DAIRY AND MEAT ANALOGUES CONTAINING E UGLENA -DERIVED

COMPONENTS

CLAIM OF PRIORITY

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63/163,071, filed March 19, 2021, and to U.S. Provisional Application No. 63/163,072, filed March 19, 2021, which are incorporated herein by reference in their entirety.

FIELD

[0002] Embodiments herein are directed to dairy and meat analogue compositions and their use in a food products, wherein the meat and dairy analogues possess one or more functional properties similar to a natural dairy product. Embodiments herein are directed to methods of making dairy and meat analogues.

BACKGROUND

[0003] As a civilization we face significant challenges in the years ahead. The population growth predicted over the next few decades, about 9.7 billion global population by 2050, will cause severe food shortages. Malnutrition is a leading cause of death, accounting for about 3.5 million deaths per year. Global deforestation will cause the loss of a significant portion of the food we rely on currently, i.e. palm oil. The resources we utilize currently are unsustainable, two planets worth of resources will be needed to support the expected population by 2050. Accordingly, there is a significant need to identify sustainable alternatives. For example, food sources that can provide improved functionality, higher nutritional value, minimal waste stream, reduced water usages and reduced carbon dioxide emissions.

[0004] Microalgae are a rich source of protein, essential fatty acids, vitamins, and minerals. After lipid removal, the residual biomass contains even higher concentrations of protein and other nutrients. Microalgae are good sources of long chain polyunsaturated fatty acids (“PUFA”) and have been used to enrich diets with omega-3 PUFA. Described herein are, inter alia, novel techniques for extracting a variety of components from heterotrophically cultivated microalgae, e.g., Euglena, without the use of harsh chemicals or solvents.

[0005] A specific species of algae named Euglena gracilis (hereinafter Euglena or Euglena) belongs to a group of single-celled microscopic algae, that is often used as a candidate species for laboratory studies and technological applications. Euglena possess the representative features typical of eukaryotic cells such as a mitochondria, nucleus, and lysosome. Euglena can further be characterized for its long flagellum and large red eyespot. They are distinctive as they can produce their own nourishment (autotrophic) similar to plants, as well as eat and digest external food sources (heterotrophic) like animals. Euglena is a demonstrated, multifaceted model organism for study. Through optimizing the natural ability to employ singly or both modes of nourishment, Euglena can be directed to produce target compounds by adjusting key parameters in the production process. These critical adjustments can be used to enhance the natural mechanisms of the microorganism, to encourage rapid growth and the efficient conversion of valuable products with little waste production.

[0006] Dairy is a staple ingredient, found in most homes worldwide. As the population continues to lean towards a healthier and more sustainable lifestyle, this has led to a need in the marketplace for a sustainable, healthy dairy alternative. The disclosed dairy analogues are versatile affordable complete nutritive solution. Currently available vegan and plant based dairy substitutes lack full nutritional benefits as well as essential amino acids, creating an incomplete nutritive product. Further, the commercially available dairy substitute solutions lack the texture, flavor, and color of the commonly used dairy products.

[0007] Meat is a staple ingredient, found in most homes worldwide. As the population continues to lean towards a healthier and more sustainable lifestyle, this has led to a need in the marketplace for a sustainable, healthy meat alternative. The disclosed meat analogue is a versatile affordable complete nutritive solution. There are very few meat analogues available on the market. Further, the commercially available meat substitute solutions lack the texture, flavor, and color of the commonly used meat. Manufacturers of these products must add artificial colors or additional ingredients to mimic the natural look and feel of meat. There has been widespread interest in a suitable replacement for meat for quite some time, due to spoilage, concern for animal welfare, environmental damage from the industrial meat.

SUMMARY

[0008] The disclosure provides a dairy analogue composition. The dairy analogue compositions comprises about 1% to about 99.5% W/WEuglena-derved material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0009] Some embodiments of the present disclosure are directed to a food product comprising a dairy analogue composition comprising about 1% to about 99.5% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0010] Some embodiments of the present disclosure are directed to a method of making a dairy analogue, wherein the dairy analogue is selected from a yogurt, a butter, a creamer, a cream cheese, a cheese sauce, and a cheese.

[0011] Some embodiments of the present disclosure are directed to a food product comprising a dairy analogue composition comprising about 1% to about 99.5% W/W Euglena- derived material, a Euglena beta glucan isolate emulsion, and one or more additional ingredients, wherein the dairy analogue food product comprises one or more functional properties of a natural dairy product.

[0012] Some embodiments of the present disclosure are directed to a method of making a Euglena beta glucan isolate emulsion.

[0013] Embodiments described herein are directed to a meat analogue composition comprising about 0.5% to about 95% W/W Euglena-derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural animal meat product.

[00141 Some embodiments of the present disclosure are directed to a food product comprising a meat analogue composition comprising about 0.5% to about 95% W/W Euglena- derived material and one or more additional ingredient, wherein the meat analogue composition comprises one or more functional properties of a natural animal meat product.

[0015] Some embodiments of the present disclosure are directed to a method of making a texturized protein comprising Euglena flour.

BRIEF DESCRIPTION OF THE DRAWINGS:

[0016] FIG. 1 is a photograph of a Euglena- derived vegan yogurt formulation.

[0017] FIG. 2A is a photograph of a Euglena- derived beta-glucan isolate (“BGI”) based yogurt formulation.

[0018] FIG. 2B is a photograph of a Euglena- derived RTG based yogurt formulation.

[0019] FIG. 3 is a photograph of a Euglena- derived vegan butter spread formulation.

[0020] FIG. 4A is a photograph of a Euglena- derived BGI based cream cheese formulation.

[0021] FIG. 4B is a photograph of a ready to gel based cream cheese formulation. [0022] FIG. 5A is a photograph of a Euglena- derived creamer formulation before it was added to coffee.

[0023] FIG. 5B is a photograph of a Euglena- derived creamer formulation, after it was added to coffee.

|0024] FIG. 6 is a photograph of a Euglena- derived cheese in shells and cheese.

[0025] FIG. 7 is a photograph of the texture of Vegan Yogurt made with Euglena

BGI.

[0026] FIG. 8 is a photograph of appearance of Vegan Cheese VI with Euglena Protein Concentrate.

[0027] FIG. 9 is a photograph of appearance of Softer Vegan Cheese, V2 with Euglena protein concentrate.

[0028] FIG. 10 is a photograph of a vegan cream cheese using Euglena BGI, formulation 1 (see Example 10), pH = 4.4.

[0029] FIG. 11 is a photograph of a vegan cream cheese using Euglena BGI, formulation 2 (see Example 10), pH = 5.28.

[0030] FIG. 12 is a photograph of a vegan cream cheese using Euglena BGI, formulation 3 (see Example 10), pH = 4.81.

[0031] FIG. 13 is a photograph of a vegan cream cheese using Euglena BGI, formulation 4 (see Example 10), pH = 4.53.

[0032] FIG. 14A is a schematic representation of a preparation method of BGI emulsion. BGI was incubated into water overnight, and then oil was added.

[0033] FIG. 14B is a schematic representation of a preparation method of BGI emulsion. BGI was incubated into oil overnight, and then water was added.

[0034] FIG. 15 A is a microscopic image of the emulsion without BGI with 40% oil and 60% water. The emulsion was prepared by mixing oil with water without BGI, then the mixture was homogenized using a high power homogenizer at highest power (Stage 3). The microscopic image was taken with 40-60x magnification, and the scale bar represents 100 pm.

[0035] FIG. 15B is a series of microscopic images of emulsions where 10% BGI was incubated in water (60%) overnight (Stage 1) and then oil (30%) was added, and the mixture was homogenized (Stage 3). All emulsions were prepared using a high power homogenizer at highest power. The microscopic images were taken with 40-60x magnification, and the scale bar represents 100 pm. [00361 FIG. 15C is a series of microscopic images of emulsions where 10% BGI was incubated in oil (30%) overnight (Stage 1) and then water (60%) was added (Stage 2, before homogenization), then the mixture was homogenized (Stage 3). All emulsions were prepared using a high power homogenizer at highest power. The microscopic images were taken with 40-60x magnification, and the scale bar represents 100 pm.

[0037] FIG. 16A is a series of microscopic images of the BGI emulsion prepared under Vortex for 2 mins, and the BGI incubation time in oil was 24 hours, 96 hours or 10 days. Images were taken at 40-60x magnification, and the scale bar represents 50 - 100 pm.

[0038] FIG. 16B is a series of microscopic images of the BGI emulsion prepared using a Fisherbrand 150 homogenizer with a speed of 35,000rpm and homogenized for 5 mins, and the BGI incubation time in oil was 24 hours, 96 hours or 10 days. Images were taken at 40-60x magnification, and the scale bar represents 50 - 100 pm.

[0039] FIG. 16C is a series of microscopic images of the BGI emulsion prepared using an Omni international GLH-01 homogenizer with a speed of 28,000 rpm and homogenized for 5 mins, and the BGI incubation time in oil was 24 hours. Images were taken at 40-60x magnification, and the scale bar represents 50 - 100 pm.

[0040] FIG. 17 is a series of microscopic images of the effects of sample compositions on BGI emulsion formation. SBGI: Spray dried BGI; FBGI: Freeze dried BGI; Biomass: Spray dried protein rich biomass; FMP: fully milled Paramylon; PMP: partially milled Paramylon. The time in the sample label indicates the material's incubation time in oil. BGIs and Biomass have the composition of 1:3:6 (BGI or biomass: oil: water), FMP and PMP have the composition of 0.5:5: 10 (milled: oil: water). The images were taken on diluted emulsion samples with 40-60x magnification, and the scale bar represents 50 - 100 pm.

[0041] FIG. 18A is a series of microscopic images of the comparison of the top phases of the emulsions of spray dried BGI (SBGI) and freeze dried BGI (FBGI).

[0042] FIG. 18B is a series of microscopic images of the comparison of the bottom phases of spray dried BGI (SBGI) and freeze dried BGI (FBGI).

[0043] FIG. 19 is a line graph depicting oil binding capacity (g/g) of SBGI with increasing incubation time.

[0044] FIG. 20A is a microscopic image of the oil and BGI phase of a BGI emulsion made with a low power homogenizer and a short oil incubation time (Type 1, < 16 hours).

[0045] FIG. 20B is a microscopic image of the oil and BGI phase of a BGI emulsion made with a low power homogenizer and an oil incubation time of 24 hours (Type 2). [00461 FIG. 20C is a microscopic image of the oil and BGI phase of a BGI emulsion made with a low power homogenizer and an oil incubation time of 48 hours (Type 3).

[0047] FIG. 20D is a microscopic image of the oil and BGI phase of a BGI emulsion made with a low power homogenizer and an oil incubation time of a long period of time, i.e. 96 hours (Type 4).

[0048] FIG. 20E is a microscopic image of the BGI water phase of a BGI emulsion made with a low power homogenizer and a short oil incubation time (Type 1, < 16 hours).

[0049] FIG. 20F is a microscopic image of the BGI water phase of a BGI emulsion made with a low power homogenizer and an oil incubation time of 24 hours (Type 2).

[0050] FIG. 20G is a microscopic image of the BGI water phase of a BGI emulsion made with a low power homogenizer and an oil incubation time of 48 hours (Type 3).

[0051] FIG. 20H is a microscopic image of the BGI water phase of a BGI emulsion made with a low power homogenizer and an oil incubation time of a long period of time, i.e. 96 hours (Type 4).

[0052] FIG. 21 is a microscope image of an oil and water emulsion formed from 30% oil in water. Scale bar represents 100 pm.

[0053] FIG. 22A is a microscopic image showing the effect of BGI content on emulsions. The composition was a weight ratio of BGFoihwater of 5:20:80. The emulsion was prepared from spray dried BGI using a high powered homogenizer with a speed of 28,000 rpm and homogenized for 5 min. Scale bar represent 100 pm.

[0054] FIG. 22B is a microscopic image showing the effect of BGI content on emulsions. The composition was a weight ratio of BGFoihwater of 10:20:80. The emulsion was prepared from spray dried BGI using a high powered homogenizer with a speed of 28,000 rpm and homogenized for 5 min. Scale bar represent 100 pm.

[0055] FIG. 22C is a microscopic image showing the effect of BGI content on emulsions. The composition was a weight ratio of BGFoihwater of 15:20:80. The emulsion was prepared from spray dried BGI using a high powered homogenizer with a speed of 28,000 rpm and homogenized for 5 min. Scale bar represent 100 pm.

[0056] FIG. 22D is a microscopic image showing the effect of BGI content on emulsions. The composition was a weight ratio of BGFoihwater of 20:20:80. The emulsion was prepared from spray dried BGI using a high powered homogenizer with a speed of 28,000 rpm and homogenized for 5 min. Scale bar represent 100 pm. [0057] FIG. 23A is a microscopic image showing a BGI emulsion with low BGI content (0.25% BGI). The emulsion was prepared using Omni international GLH-01 with speed of 28,000rpm and homogenized for 2 min. BGI was incubated in oil for 6 days before being homogenized. The microscopic images were taken at 40X magnification, and the scale bar represents 50 pm.

[0058] FIG. 23B is a microscopic image showing a BGI emulsion with low BGI content (0.6% BGI). The emulsion was prepared using Omni international GLH-01 with speed of 28,000rpm and homogenized for 2 min. BGI was incubated in oil for 6 days before being homogenized. The microscopic images were taken at 40X magnification, and the scale bar represents 50 pm.

[0059] FIG. 23 C is a microscopic image showing a BGI emulsion with low BGI content (1.2% BGI). The emulsion was prepared using Omni international GLH-01 with speed of 28,000rpm and homogenized for 2 min. BGI was incubated in oil for 6 days before being homogenized. The microscopic images were taken at 40X magnification, and the scale bar represents 50 pm.

[0060] FIG. 23D is a microscopic image showing a BGI emulsion with low BGI content (2.5% BGI). The emulsion was prepared using Omni international GLH-01 with speed of 28,000rpm and was homogenized for 2 min. BGI was incubated in oil for 6 days before being homogenized. The microscopic images were taken at 40X magnification, and the scale bar represents 50 pm.

[0061] FIG. 24 is a series of microscopic images of emulsions with different canola oil content. Emulsion was prepared from Spray dried BGI using a high power homogenizer with speed of 28,000rpm and homogenized for 5 min, and prepared with a composition of weight ratio (BGI: oil: water). CalO: 10: 10:80; Ca20: 10:20:70; Ca30: 10:30:60; Ca40: 10:40:50; Ca45: 10:45:45; Ca60: 10:60:30; Ca70: 10:70:20.

[0062] FIG. 25A is a series of microscopic images taken at 2 hours after homogenization with different oils. The composition was BGLoikwater 10: 10:80. The emulsions were prepared using the high power homogenizer with speed of 28,000 rpm, and homogenized for 5 min. Ca: Canola oil, Co: Coconut oil, P: Palm oil, Li: Linseed oil, Su: sunflower oil. The images were taken using 40x magnification and the scale bar represents 100 pm.

[0063] FIG. 25B is a series of microscopic images taken at 2 hours after homogenization with different oils. The composition was BGLoikwater 10:45:45. The emulsions were prepared using the high power homogenizer with speed of 28,000 rpm, and homogenized for 5 min. Ca: Canola oil, Co: Coconut oil, P: Palm oil, Li: Linseeds oil, Su: sunflower oil. The images were taken using 40x magnification, and the scale bar represents 100 pm.

[0064] FIG. 26 is a series of microscopic images of Canola oil emulsions at different storage temperatures. The composition of emulsion had the composition of BGI: oil: water 10:30:60, prepared using Omni International GLH-01 homogenizer at the speed of 28,000 RPM and homogenized for 5 min. The emulsions were stored at -20°C, 4°C, and room temperature for 16 hours, and 2 hours at 80°C. Ca: Canola oil; 30 indicates oil content as 30%; temperature in the label indicates the storage temperature; the images were taken using 40x magnification, with scale bars representing 100 pm.

[0065| FIG. 27 is a series of images of the textures of the emulsions prepared using different compositions. The emulsions were prepared using Omni international GLH-01 with speed of 28,000 rpm and homogenized for 5 mins. Prior to homogenization, BGI was incubated in oil for 16 hours. The textures were physically observed. The compositions of the BGPoikwater emulsions were as follows: Watery: 0:30:70; 2% milk: 5:10:85; 3:5% cream: 5:30:65; 10% cream: 10:30:60; Drinkable yogurt: 10:40:50; Normal yogurt: 20:30:50; Greek yogurt: 20:40:40; Ricotta Cheese/Mozzarella: 30:30:40; and Bocconcini cheese: 40:40:20. The microscopic images were taken at 40x magnification and the scale bar represents 100 pm.

[0066] FIG. 28A is a photograph image of top view of coffee with BGI creamer prepared using a Kitchen Aid mixer.

[0067] FIG. 28B is a photograph image of top view of coffee with BGI creamer prepared using an Omni GLH polytron blender and commercial creamers.

[0068] FIG. 28C is a photograph image of top view of coffee with BGI creamer prepared using Nestle coconut milk based creamer.

[0069] FIG. 28D is a photograph image of top view of coffee with BGI creamer prepared using Earth’s Own oat based creamer.

[0070] FIG. 29A is a series of photograph images of coffee with creamer prepared using a polytron right after the addition of creamer to coffee (0 min) and 30 minutes after the addition of creamer to coffee (30 min).

[0071] FIG. 29B is a series of photograph images of coffee with creamer prepared using a high pressure homogenizer right after the addition of creamer to coffee (0 min) and 30 minutes after the addition of creamer to coffee (30 min). [0072] FIG. 30A is a photograph image of creamer prepared with all ingredients in the formula shown in Table 1 after storing in the refrigerator for 2 days.

[0073] FIG. 30B is a photograph image of creamer prepared without the sugar in the formula shown in Table 1 after storing in the refrigerator for 2 days.

|0074] FIG. 30C is a photograph image of creamer prepared without the sunflower lecithin in the formula shown in Table 1 after storing in the refrigerator for 2 days.

[0075] FIG. 30D is a photograph image of creamer prepared without the carrageenan in the formula shown in Table 1 after storing in the refrigerator for 2 days.

[0076] FIG. 30E is a photograph image of creamer prepared without the BGI in the formula shown in Table 1 after storing in the refrigerator for 2 days.

[0077] FIG. 30F is a photograph image of creamer prepared without the coconut oil in the formula shown in Table 1 after storing at the refrigerator for 2 days.

[0078] FIG. 31A is a photograph image of the performance of BGI creamers without any protein and with no gum additives.

[0079] FIG. 3 IB is a photograph image of the performance of BGI creamers without any protein and with no guar gum.

[0080] FIG. 31C is a photograph image of the performance of BGI creamers without any protein and with no acacia gum or gellan gum.

[0081] FIG. 3 ID is a photograph image of the performance of BGI creamers without any protein and with no carrageenan.

[0082] FIG. 32 is a photograph image of the emulsion prepared using wet ready to gel beta glucan isolate for yogurt application right after preparation.

[0083] FIG. 33 A is a photograph image of cream cheese developed using dairy formula 9 (see Table 35). Oat milk flour is the protein source in dairy formula 9.

[0084] FIG. 33B is a photograph image of cream cheese developed using dairy formula 11 ( see Table 35). Pea protein is the source of protein in dairy formula 11.

[0085] FIG. 34A is a representative image of cream cheese analogue prepared using

BGI with Admul as an emulsifier.

[0086] FIG. 34B is a representative image of cream cheese analogue prepared using BGI with Myverol 19-02K as an emulsifier.

[0087] FIG. 35 A is a photograph of a Euglena- derived textured vegetable protein (“TVP”) formulation.

[0088] FIG. 35B is a photograph of a Euglena- derived high moisture extrusion cooking (“HMEC”) formulation. [0089] FIG. 36A is a photograph of a bacon-like product prepared using a HMEC process formulation.

[0090] FIG. 36B is a photograph of a pulled pork-like product prepared using a HMEC process formulation.

|0091] FIG. 37A is a photograph of a Euglena-dcnvcd chicken-like product in a chicken soup.

[0092] FIG. 37B is a photograph of a Thai chicken and rice soup.

[0093] FIG. 38 is a photograph of finely flaked extruded Euglena- derived tuna-like product made into a wrap.

[0094] FIG. 39A is a photograph of a Euglena- derived protein crisps food product.

[0095] FIG. 39B is a photograph of a Euglena- derived protein crisps food product.

[0096] FIG. 40 is a photograph of prepared and baked Chicken Nugget prepared with Euglena TVP.

[0097] FIG. 41 is a photograph of prepared Euglena TVP Sausage Link.

[0098] FIG. 42 is a photograph of prepared Pulled Pork with Euglena TVP.

[0099] FIG. 43 is a photograph of prepared Fish Stick with Euglena TVP.

[0100] FIG. 44 is a photograph of prepared Italian Dinner Sausage with Euglena

TVP.

[0101] FIG. 45 is a photograph of prepared Burger Patty with Euglena TVP.

[0102] FIG. 46A is a series of images of the different HMMA formulas that show the tear-ability of the product. 1) Formulation with wet Euglena biomass and Pea Protein Isolate 2. 2) Formulation with wet Euglena biomass and Pea Protein Isolate 1. 3) Formulation with Euglena flour (15%) and Pea Protein Isolate 1 4) Formulation with Euglena flour (10%) and Pea Protein Isolate 1.

[0103] FIG. 46B is a series of images of the different HMMA formulas that show the product. 1) Formulation with wet Euglena biomass and Pea Protein Isolate 2. 2) Formulation with wet Euglena biomass and Pea Protein Isolate 1. 3) Formulation with Euglena flour (15%) and Pea Protein Isolate 1 4) Formulation with Euglena flour (10%) and Pea Protein Isolate 1.

[0104] FIG. 47A is a photograph of the textured protein from TVP Formula 1, representing a Chunk product at a slower speed.

[0105] FIG. 47B is a photograph of the texturized protein from TVP Formula 1, representing a Die Cut product at a higher speed. [0106] FIG. 47C is a photograph of the texturized protein representing an Urshel comitrol treated product.

[0107] FIG. 48A is a photograph of the texturized protein from TVP Formula 2, representing a Chunk product at a slower speed.

|0108] FIG. 48B is a photograph of the texturized protein from TVP Formula 2, representing a Die Cut product at a higher speed.

[0109] FIG. 48C is a photograph of the texturized protein from TVP Formula 2, representing an Urshel comitrol treated product.

DETAILED DESCRIPTION

[0110] This invention is not limited to the particular processes, compositions, or methodologies described, as these may vary. The terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. All publications mentioned herein, are incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

Definitions

[0111 ] For convenience, certain terms employed in the specification, examples and claims are collected here. Unless defined otherwise, all technical and scientific terms used in this disclosure have the same meanings as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0112] In the present disclosure, where a range of values is provided, it is intended that each intervening value between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the disclosure. For example, if a range of 1 g to 8 g is stated, it is intended that 2 g, 3 g, 4 g, 5 g, 6 g, and 7 g are also explicitly disclosed, as well as the range of values greater than or equal to 1 g and the range of values less than or equal to 8 g. [01131 The singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a “cell” includes a single cell as well as two or more of the same or different types of cells.

[0114] The word “about” when immediately preceding a numerical value means a range of plus or minus 5% of that value, e.g, “about 50” means 45 to 55, “about 25,000” means 22,500 to 27,500, etc., unless the context of the disclosure indicates otherwise, or is inconsistent with such an interpretation. Furthermore, the phrases “less than about” a value or “greater than about” a value should be understood in view of the definition of the term “about” provided herein.

[0115] “Baked good” means a food item, typically found in a bakery, that is prepared by using an oven and usually contain a leavening agent. Baked goods include, but are not limited to breads, brownies, cookies, pies, cakes and pastries.

[0116] As used herein “beta glucan isolate” or “BGI” refers to beta-l,3-D-glucan that has been extracted from Euglena cells and is a stored polysaccharide. In Euglena, the terms “beta glucan” and “paramylon” can be used interchangeably. “Beta glucan” is stored as small discoid granules between 1 to 2 pm in size and is a high molecular weight and high crystallinity polymer b-glucan is a long-chain polysaccharide made of glucose monomers connected through glycosidic linkages. It is a structural component in fungi, algae, bacteria, and plants, including cereal grains such as oats, barley, wheat, and rye. Significant quantities of b-glucan are found in barley (2% to 20%), oat (3% to 8%), and sorghum (1.1% to 6.2%). Microalgae Euglena gracilis contains about 25% to 60% (w/w) of water-insoluble linear b- 1,3 -glucan, which is also known as paramylon. Euglena gracilis produces paramylon as a fibrillar high molecular weight polymer of greater than 500 kDa. The crystallinity of paramylon in its native state approaches 90%. In Euglena cells, paramylon is deposited as granules (~1 to 2 pm) that corresponds to 100% glucose in NMR spectra. Isolated paramylon is a fine, free flowing white powder.

[0117] “Bread” means a food item that contains flour, liquid, and usually a leavening agent. Breads are usually prepared by baking in an oven, although other methods of cooking are also acceptable. The leavening agent can be chemical or organic/biological in nature. Typically, the organic leavening agent is yeast. In the case where the leavening agent is chemical in nature (such as baking powder and/or baking soda), these food products are referred to as “quick breads.” Crackers and other cracker-like products are examples of breads that do not contain a leavening agent. [01181 The transitional term “comprising,” which is synonymous with “including,” “containing,” or “characterized by,” is inclusive or open-ended and does not exclude additional, un-recited elements or method steps. By contrast, the transitional phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim. The transitional phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps “and those that do not materially affect the basic and novel characteristic(s)” of the claimed invention. In embodiments or claims where the term comprising is used as the transition phrase, such embodiments can also be envisioned with replacement of the term “comprising” with the terms “consisting of’ or “consisting essentially of.”

[0119] “Dairy analogue” as used herein refers to a Euglena-derived product that has a resemblance to dairy products.

[0120] “Dispersion” refers to a distribution of particles more or less evenly throughout a medium, including a liquid or gas. One common form of dispersion is an emulsion made up of a mixture of two or more immiscible liquids such as oil and water.

[0121] The term “dry biomass” as used herein refers to biomass or Euglena biomass that exists in the absence of water.

[0122] “Dry weight” means weight determined in the relative absence of water. For example, reference to a dry mixture refers to a specified percentage of a particular component(s) by dry weight as a percentage and is calculated based on the weight of the composition before any liquid has been added.

[0123] As used in this disclosure, the terms “emulsifying,” “emulsion” or a derivative thereof refers to where a substance is present in a food composition or food product as a single-phase mixture where a two-phase system of oil and water would normally have existed. An emulsion thus refers to a kinetically stable mixture of two normally immiscible liquids, i.e. oil and water. In some other foods, the water is dispersed in oil.

[0124] “Edible ingredient” means any substance or composition which is fit to be eaten. “Edible ingredients” include, without limitation, grains, fruits, vegetables, proteins, herbs, spices, carbohydrates, and fats.

[0125] As used herein, the term “enriched” refers to an increase in a percent amount of a molecule, for example, a protein, in one sample relative to the percent amount of the molecule in a reference sample. In some embodiments, the enrichment is on a weight to weight basis. In some embodiments, the enrichment refers to an increase of about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% relative to the reference value or amount. In some embodiments, the enrichment refers to an increase of at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% relative to the reference value or amount.

[0126] The term uglena -derived material” as used herein refers to any material derived from the species of Euglena. “Euglena- derived material” can be Euglena biomass (e.g., fresh, wet or dry biomass), a Euglena derived protein (i.e., Euglena protein), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate, a Euglena protein isolate, a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, ready to gel Euglena beta-glucan isolate, Euglena beta- glucan slurry, a Euglena oil, milled Euglena paramylon, Euglena wet protein concentrate, and combinations thereof. Euglena is a genus of single cell flagellate eukaryotes. Euglena gracilis is a freshwater species of single-celled alga in the genus Euglena. It has secondary chloroplasts and is a mixotroph able to feed by photosynthesis or phagocytosis. The process of obtaining Euglena- derived materials are described in WO2020/261245, which is incorporated herein by reference in its entirety.

[0127] “Finished food product” and “finished food ingredient” mean a food composition that is ready for packaging, use, or consumption. For example, a “finished food product” may have been cooked or the ingredients comprising the “finished food product” may have been mixed or otherwise integrated with one another. A “finished food ingredient” is typically used in combination with other ingredients to form a food product.

[0128] The term “functional food product” as used herein refers to a food product given an additional function by adding new ingredients or more of existing ingredients. For example, where protein is added to a food product to provide texture, water holding capacity or nutritional support to a food product.

[0129] “Food,” “food composition,” “food product,” and “foodstuff’ mean any composition intended to be or expected to be ingested by humans or other animals as a source of nutrition and/or calories. Food compositions are composed primarily of carbohydrates, fats, water and/or proteins and make up substantially all of an individual’s daily caloric intake.

[0130] “Foamability” as used herein, refers to the ability of a material to rapidly adsorb on the air-liquid interface during whipping or bubbling, and to form a cohesive viscoelastic film by way of intermolecular interactions.

[0131] The term “fresh biomass” or “fresh Euglena biomass” as used herein refers to biomass or Euglena biomass that is not frozen or dried after harvesting and is kept at 4 °C until use. [0132] The term “gelling,” “gelification” or a derivative thereof as used herein refers to a food composition or a food product in a gelatinous form. A gelatinous form is created by incorporating solids and liquids into a uniform three dimensional, semi-solid structure. A gelatinous food product is considered a soft gel when its tensile strength is in the range of 500 1000 g/cm ² , as seen in, for example, jelly and jams, nut butters (e.g. just nuts versions), jelly- like products, and fondant. A gelatinous food product is considered a hard gel when its tensile strength is in the range of 1000-3000 g/cm ² , as seen in, for example, gummy candy, confectionary gels (i.e. cookie filling), fruit gel bars, and fruit snacks.

[0133] “Good manufacturing practice” and “GMP” mean those conditions established by regulations set forth at 21 C.F.R. 110 (for human food) and 111 (for dietary supplements), or comparable regulatory schemes established in locales outside the United States. The U.S. regulations are promulgated by the U.S. Food and Drug Administration under the authority of the Federal Food, Drug, and Cosmetic Act to regulate manufacturers, processors, and packagers of food products and dietary supplements for human consumption. All of the processes described herein can be performed in accordance with GMP or equivalent regulations. In the United States, GMP regulations for manufacturing, packing, or holding human food are codified at 21 C.F.R. 110 These provisions, as well as ancillary provisions referenced therein, are hereby incorporated by reference in their entirety for all purposes. GMP conditions in the Unites States, and equivalent conditions in other jurisdictions, apply in determining whether a food is adulterated (the food has been manufactured under such conditions that it is unfit for food) or has been prepared, packed, or held under unsanitary conditions such that it may have become contaminated or otherwise may have been rendered injurious to health. GMP conditions can include adhering to regulations governing: disease control; cleanliness and training of personnel; maintenance and sanitary operation of buildings and facilities; provision of adequate sanitary facilities and accommodations; design, construction, maintenance, and cleanliness of equipment and utensils; provision of appropriate quality control procedures to ensure all reasonable precautions are taken in receiving, inspecting, transporting, segregating, preparing, manufacturing, packaging, and storing food products according to adequate sanitation principles to prevent contamination from any source; and storage and transportation of finished food under conditions that will protect food against physical, chemical, or undesirable microbial contamination, as well as against deterioration of the food and the container. [0134] The term “low moisture meat analogue” as used herein refers to a meat analogue that has been developed through a low moisture extrusion process to texturized the protein into a meat analogue.

[0135] The term “high moisture meat analogue” as used herein refers to a meat analogue that has been developed through a high moisture extrusion process in order to texturize the protein into a fibrous and meat-like structure. It is better producing the fibrous texture in the meat analogue over low moisture extrusion.

[0136] “Increased lipid yield” means an increase in the lipid/oil productivity of a microalgal culture that can achieved by, for example, increasing the dry weight of cells per liter of culture, increasing the percentage of cells that contain lipid, and/or increasing the overall amount of lipid per liter of culture volume per unit time.

[0137] “Milk alternative” as used herein refers to any substance that resembles milk and can be used in the same ways as an animal-derived milk. The milk alternative can be in liquid or dry powder form. A milk alternative can be a nut milk, coconut milk, or a grain milk such as oat milk.

[0138] “Meat analogue” as used herein refers to a Euglena- derived product that has a resemblance to meat products.

[0139] The terms “natural animal meat product” and “natural animal dairy product” as used herein refer to meat and dairy products, respectively, that are derived from animals.

[0140] The term “Oil” as used herein means any triacylglycerol (or triglyceride oil), produced by organisms, including microalgae, other plants, and/or animals. “Oil,” as distinguished from “fat,” refers, unless otherwise indicated, to lipids that are generally liquid at ordinary room temperatures and pressures. For example, “oil” includes vegetable or seed oils derived from plants, including without limitation, an oil derived from soy, rapeseed, grapeseed, canola, palm, palm kernel, coconut, com, olive, sunflower, cotton seed, cuphea, peanut, camelina sativa, mustard seed, cashew nut, oats, lupine, kenaf, calendula, hemp, coffee, linseed, hazelnut, euphorbia, pumpkin seed, coriander, camellia, sesame, safflower, rice, tung oil tree, cocoa, copra, opium poppy, castor beans, pecan, jojoba, jatropha, macadamia, Brazil nuts, and avocado, Euglena derived oil, medium-chain triglycerides (MCT), palmitic, omega- 3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and oleic acid as well as combinations thereof. “Oil” means any triacylglyceride (or triglyceride), produced by organisms, including microalgae, other plants, and/or animals. “Oil,” as distinguished from “fat,” refers, unless otherwise indicated, to lipids that are generally liquid at ordinary room temperatures and pressures. [0141] The term “shaking” as used herein refers to the movement of a sample, in an up and down or side to side, rapid, forceful or jerky movement. This may be done manually, or mechanically.

[0142] The term “solution” as used herein refers to a homogeneous mixture of a substance (solute) dispersed through a liquid medium (solvent) that cannot be separated by the forces of gravity alone.

[0143] The term “solid content” as used herein refers to how much mass (i.e. biomass or protein concentrate) is in a liquid by weight of each.

[0144] The term “substantially free” as used herein refers to the complete or near complete lack of light or a component. For example, a composition that is “substantially free” of water would either completely lack water, or so nearly completely lack water that the effect would be the same as if it completely lacked water.

[0145] As used herein, the term “stability” and derivatives thereof refer to heat stability, freeze-thaw stability, light stability, emulsion stability, or storage stability. Heat stability is the ability of a product or material to retain the same properties after exposure to a high heat for a single set period of time or a cycling of exposure times. Freeze-thaw stability is the ability of a product or material to retain the same properties after being frozen and subsequently thawed, which can be cycled through a number of freeze thaw cycles. Light stability is the ability of a product or material to retain the same properties after exposure to a light, such as sunlight or indoor light for a single set period of time or a cycling of exposure times. Emulsion stability is the ability of a product or material to retain an emulsion and to prevent separating, over time. Further, the term “stabilizer” relates to a material that provides stability described herein when added to a product or another material . For example, a stabilizer may be an ingredient incorporated into a final food formulation which preserves the structure and sensory characteristics of a food product over time, which would not otherwise be maintained in the absence of the stabilizer.

[0146] The term “solubility” as used herein, refers to the maximum amount of a substance that is able to be completely dissolved in a solution, usually in a specific amount.

[0147] “Uncooked product” means a composition that has not been subjected to heating but may include one or more components previously subjected to heating.

[0148] As used herein, the term “viscosity” refers to the resistance of a fluid when attempting to flow, may also be thought of as a measure of fluid friction.

[0149] The term “wet biomass” as used herein refers to biomass or Euglena biomass comprising moisture. A “wet biomass” may be a solid, gel or liquid. [0150] “W/W” or “w/w,” in reference to proportions by weight, means the ratio of the weight of one substance in a composition to the weight of the composition. For example, reference to a composition that comprises 5% w/w microalgal biomass means that 5% of the composition’s weight is composed of microalgal biomass (e.g., such a composition having a weight of 100 mg would contain 5 mg of microalgal biomass) and the remainder of the weight of the composition (e.g., 95 mg in the example) is composed of other ingredients.

[0151] “W V” or “w/v” means the ratio of the weight of one substance in a composition to total volume of the composition. For example, reference to a composition that comprises 5% w/v microalgal biomass means that 5g of microalgal biomass is dissolved in a final volume of 100 mL aqueous solution.

[0152] The term “whipping” as used herein, refers to the action of using a whisk, or a mixer to beat a sample in order to rapidly incorporate air and produce expansion.

[0153] The term “water holding capacity” or WHC or a derivative thereof as used herein relating to food composition or product refers to the ability to hold the food’s own and added water during the application of forces, pressing, centrifugation, or heating. WHC may also be described as a physical property, for example, the ability of a food structure to prevent water from being released from the three-dimensional structure of, for example, a gel.

[0154] Throughout this disclosure, the Euglena may be selected from the group of species selected from Euglena gracilis, Euglena sanguinea, Euglena deses, Euglena mutabilis, Euglena acus, Euglena virdis, Euglena anabaena, Euglena geniculata, Euglena oxyuris, Euglena proxima, Euglena tripteris, Euglena chlamydophora, Euglena splendens, Euglena texta, Euglena intermedia, Euglena polymorpha, Euglena ehrenbergii, Euglena adhaerens, Euglena clara, Euglena elongata, Euglena elastica, Euglena oblonga, Euglena pisciformis, Euglena cantabrica, Euglena granulata, Euglena obtusa, Euglena limnophila, Euglena hemichromata, Euglena variabilis, Euglena caudata, Euglena minima, Euglena communis, Euglena magnifica, Euglena terricola, Euglena velata, Euglena repulsans, Euglena clavata, Euglena lata, Euglena tuberculata, Euglena contabrica, Euglena ascusformis, Euglena ostendensis, Chlorella autotrophica, Chlorella colonials, Chlorella lewinii, Chlorella minutissima, Chlorella pituita, Chlorella pulchelloides, Chlorella pyrenoidosa, Chlorella rotunda, Chlorella singularis, Chlorella sorokiniana, Chlorella variabilis, Chlorella volutis, Chlorella vulgaris, Schizochytrium aggregatum, Schizochytrium limacinum, Schizochytrium minutum, and combinations thereof. In certain embodiments, the microalgae is Euglena gracilis. [01551 Various aspects now will be described more fully hereinafter. Such aspects may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey its scope to those skilled in the art.

Dairy Analogue Composition

[0156] Embodiments described herein are directed to a dairy analogue composition comprising about l% to about 100% W/W Euglena-dcnvcd material (e.g., about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%), and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0157J In some embodiments described herein, the dairy analogue composition comprises about l%to about 100% W/W Euglena-dcnvcd material, about 0.05%to about 70% W/W additional protein source, and one or more additional ingredient. In some embodiments, the dairy composition comprises about 10% to about 98% W/W Euglena- derived material. In some embodiments, the dairy analogue composition comprises about 10% to about 98% W/W Euglena- derived material. In some embodiments, the dairy analogue composition comprises about 0.01% to about 100% W/W Euglena-dcm cd material. In some embodiments, the dairy analogue composition comprises about 0.01%, about 0.05%, about 1%, about 2%, about 3%, about 4% about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100% W/W, or any range within these values.

[0158] In embodiments described herein, the Euglena- derived material can be Euglena biomass (e.g., fresh, wet or dry biomass), a Euglena- derived protein (i.e., Euglena protein, e.g. wet or dry), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate (e.g. wet or dry), a Euglena protein isolate (e.g. wet or dry), a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, Ready To Gel Euglena beta glucan isolate, Euglena beta glucan slurry, a Euglena oil, milled Euglena paramylon (e.g. wet or dry), Euglena wet protein concentrate, Euglena dry protein concentrate, and combinations thereof.

[0159] In some embodiments of the dairy analogue described herein, the fresh Euglena- derived material can be Euglena biomass (e.g., fresh, wet or dry biomass), a Euglena derived protein (i.e., Euglena protein), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate, a. Euglena protein isolate, a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, ready to gel Euglena beta-glucan isolate, Euglena beta-glucan slurry, a Euglena oil, milled Euglena paramylon, Euglena wet protein concentrate, and combinations thereof. Euglenid protozoans (e.g., Euglena gracilis) are a source of an insoluble, linear (l,3)- -glucan of high molecular mass called paramylon that occurs naturally in a high crystalline form in discrete membrane- bound granules in the cytoplasm.

[0160] In some embodiments, a dairy analogue composition comprising about 10 to about 20% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0161] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0162] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0163] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0164] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0165] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0166] In some embodiments, a dairy analogue composition comprising about 20 to about 30% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product. [0167] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0168] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0169] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0170] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0171] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0172] In some embodiments, a dairy analogue composition comprising about 30 to about 40% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0173] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0174] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0175] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0176] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0177] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0178] In some embodiments, a dairy analogue composition comprising about 40 to about 50% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0179] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0180] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0181] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0182] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0183] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [0184] In some embodiments, a dairy analogue composition comprising about 50 to about 60% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

|0185] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0186] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0187] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0188] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0189] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0190] In some embodiments, a dairy analogue composition comprising about 60 to about 70% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0191] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0192] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0193] hi some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0194] hi some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0195] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0196] In some embodiments, a dairy analogue composition comprising about 70 to about 80% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0197] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0198] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1%to about 99.9% W/W ofthe composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0199] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0200] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [02011 In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0202] In some embodiments, a dairy analogue composition comprising about 80 to about 90% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0203] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0204] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0205] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0206] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0207] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0208] In some embodiments, a dairy analogue composition comprising about 90 to about 99.5% W/W hug/ena-dcmcd material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0209] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0210] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0211] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0212] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0213] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0214] In some embodiments, a dairy analogue composition comprising about 5 to about 80% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0215] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0216] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0217] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [02181 In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0219] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0220] In some embodiments, a dairy analogue composition comprising about 10 to about 90% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0221] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0222] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0223] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0224] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0225] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0226] In some embodiments, a dairy analogue composition comprising about 20 to about 50% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0227] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0228] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0229] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0230] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0231] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0232] In some embodiments, the dairy analogue can include about 0.01% to about 100% Euglena protein concentrate. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, optionally about 3% to about 50%, or optionally about 0.5% to about 15% Euglena protein concentrate.

[0233] In some embodiments, the dairy analogue can include about 0.01% to about 100% Euglena protein flour or wet biomass. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, optionally about 0.1% to about 60%, or optionally about 5% to about 30% Euglena protein flour or wet biomass.

[0234] In some embodiments, the dairy analogue can include about 0.01% to about 100 % Euglena protein isolate. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena protein isolate. [02351 In some embodiments, the dairy analogue can include about 0.01% to about 100% Euglena beta glucan isolate. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, or optionally about l%to about 50 % Euglena beta glucan isolate.

|0236] In some embodiments, the dairy analogue can include about 0.01% to about 100% Euglena textured vegetable protein. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena textured vegetable protein. In some embodiments, the dairy analogue can include optionally about 5 to about 50% Euglena textured vegetable protein.

[0237] In some embodiments, the dairy analogue can include about 0.01% to about 100% Euglena high moisture extrusion product. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena high moisture extrusion product.

[0238] In some embodiments, the dairy analogue can include about 0.01% to about 100% Euglena low moisture extrusion product. In some embodiments, the dairy analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena low moisture extrusion product.

[0239] In some embodiments, the Euglena- derived wet protein concentrate is in an amount of about 5% to about 95%, about 15% to about 95%, about 20% to about 95%, about 25% to about 95%, about 30% to about 95%, about 30% to about 95%, about 35% to about 95%, about 40% to about 95%, about 45% to about 95%, about 50% to about 95%, about 55% to about 95% W/W, in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 94% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 56% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 7% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 40% to about 87% in a liquid mixture. In certain embodiments, the Euglena-dcnvcd wet protein concentrate is in an amount of about 40% to about 90% in a liquid mixture.

[0240] In some embodiments, the Euglena- derived wet protein concentrate is about 25% to about 80% protein, about 30% to about 80% protein, about 35% to about 80% protein, about 40% to about 80% protein, about 45% to about 80% protein, about 50% to about 80% protein, about 55% to about 80% protein, about 60% to about 80% protein, about 65% to about 80% protein, or about 25% to about 75% protein. In certain embodiments, the Euglena- derived wet protein concentrate is about 70% protein.

[0241] In some embodiments, the Euglena- derived wet protein concentrate is an Euglena- derived protein isolate.

|0242] In some embodiments, the Euglena protein concentrate can be about 5% to about 50% ofthe composition. The amount of Euglena protein concentrate can vary depending on the type of composition. In some embodiments, the Euglena protein concentrate can be wet Euglena protein concentrate. In some embodiments, the Euglena protein concentrate can be dry Euglena protein concentrate. In some embodiments there the ratio of moisture content to protein is 2: 1. In some embodiments, the moisture content of protein concentrate is about 50% to about 90%. In some embodiments, the moisture content of protein concentrate is about 70 to about 90%.

[0243] In some embodiments, the protein concentrate is a ready to gel beta glucan isolate, wherein the protein concentrate is in an amount of about 0.01% to about 100%. In certain embodiments, the protein concentrate is a Ready To Gel beta glucan isolate, wherein the protein concentrate is in an amount of about 0.1% to about 20%.

[02441 In some embodiments, the protein concentrate is a beta glucan slurry or a milled paramylon, wherein the protein concentrate is in an amount of about 0.01% to about 100%. In some embodiments, the protein concentrate is a beta glucan slurry or a milled paramylon, wherein the protein concentrate is in an amount of about 0.1% to about 100 %. In some embodiments, the protein concentrate is a beta glucan slurry or a milled paramylon, wherein the protein concentrate is in an amount of about 5% to about 95% W/W.

[0245] In some embodiments, the Euglena- derived dry protein concentrate is in an amount of about 5% to about 95%, about 15% to about 95%, about 20% to about 95%, about 25% to about 95%, about 30% to about 95%, about 30% to about 95%, about 35% to about 95%, about 40% to about 95%, about 45% to about 95%, about 50% to about 95%, about 55% to about 95% W/W, in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 94% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 56% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 7% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 87% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 90% in a liquid mixture. [02461 In some embodiments, the Euglena- derived dry protein concentrate is in an amount of about 5% to about 95%, about 15% to about 95%, about 20% to about 95%, about 25% to about 95%, about 30% to about 95%, about 30% to about 95%, about 35% to about 95%, about 40% to about 95%, about 45% to about 95%, about 50% to about 95%, about 55% to about 95% W/W, in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 94% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 56% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 7% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 87% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 90% in a dry mixture.

[0247J In some embodiments, the Euglena- derived dry protein concentrate is about 25% to about 80% protein, about 30% to about 80% protein, about 35% to about 80% protein, about 40% to about 80% protein, about 45% to about 80% protein, about 50% to about 80% protein, about 55% to about 80% protein, about 60% to about 80% protein, about 65% to about 80% protein, or about 25% to about 75% protein. In certain embodiments, the Euglena- derived wet protein concentrate is about 70% protein.

[0248] In some embodiments, the Euglena- derived dry protein concentrate is an Euglena-dcnvcd protein isolate.

[0249] For the dairy analogue using fresh biomass or wet or powdered Euglena protein concentrate maskers are required to mask the marine flavours of Euglena. An ideal masker should completely mask the off notes (both aroma and flavour) coming from Euglena during cooking and in cooked products without introducing any new flavours or aroma. Flavours can also be used to improve the flavour. Ideal flavour should provide a flavour that is similar to cooked real dairy. In certain embodiments, the maskers can be purchased from Firmenich, IFF, Givaudan, Symrise, Mane, Fona, Flavor producer, McCormick, edlong, T Hasegawa. Sensient Flavors, Robertet SA, Prova, Wild/ADM, Takasago, Synergy, and others.

[0250] In some embodiments of the dairy analogue composition described herein, the Euglena- derived material is a dry material. In some embodiments of the dairy analogue composition described herein, the Euglena- derived material is a wet material.

[0251] In some embodiments of the dairy analogue composition described herein, the Euglena- derived material can be can be Euglena biomass (e.g., fresh, wet or dry biomass), a Engle na-Ac ri ved protein (i.e., Euglena protein, e.g. wet or dry), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate (e.g. wet or dry), a Euglena protein isolate (e.g. wet or dry), a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, Ready To Gel Euglena beta glucan isolate, Euglena beta glucan slurry, a. Euglena oil, milled Euglena paramylon (e.g. wet or dry), Euglena wet protein concentrate, Euglena dry protein concentrate, and combinations thereof.

[0252] In some embodiments of the dairy analogue composition described herein, the Euglena- derived material is in an amount of about 5% to about 90%, about 50% to about 85%, about 50% to about 75%, about 50% to about 65%, about 60% to about 85%, or about 70% to about 85% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 55% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 62% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 7% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 40% to about 87% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 40% to about 90% in the mixture.

[0253] In some embodiments of the dairy analogue composition described herein, the additional protein source can be of pea protein, soy protein, com protein, wheat protein, rice protein, beans protein, seed protein, nut protein, almond protein, peanut protein, seitan protein, lentil protein, chickpea protein, flaxseed protein, wild rice protein, quom protein, chia seed protein, quinoa protein, oat protein, fava bean protein, buckwheat protein, bulgar protein, sorghum protein, millet protein, microalgae protein, yellow pea protein, mung bean protein, hemp protein, sunflower protein, canola protein, lupin protein, legumes protein, potato protein, and combinations thereof. The additional protein source may be any reasonable flour, protein concentrate, protein isolate source, and combinations thereof.

[0254] In some embodiments, the dairy analogue can include about 0.1% to about 95% W/W pea protein. In some embodiments, the dairy analogue can include optionally about 1% to about 95%, or optionally about 1% to about 75% W/W pea protein.

[0255] In some embodiments of the dairy analogue composition described herein, the additional protein source is in an amount of about 0.05% to about 70%, about 0.5% to about 70%, about l%to about 70%, about 5% to about 70%, about 10% to about 70%, about 15% to about 70%, about 0.05% to about 60%, about 0.05% to about 50%, about 0.05% to about 40%, about 0.05% to about 30%, or about 0.05% to about 20%. In certain embodiments of the dairy analogue composition described herein, the additional protein source is in an amount of about 0.05% to about 15%. In certain embodiments of the dairy analogue composition described herein, the additional protein source is in an amount of about 20% to about 60%.

[0256] In some embodiments of the dairy analogue composition described herein, the one or more additional ingredients are selected from a protein, a dietary fiber, a fat, a sugar, a sensory peptide or amino acid, or combinations thereof. In some embodiments of the dairy analogue composition described herein, the one or more additional ingredient can be gellan gum, methylcellulose, yeast extract, flavoring, antioxidant blend, maskers, leavening agents, baking powder, baking soda, enzymes, transglutaminase, emulsifiers, lecithin, mono-and diglycerides, binders, carrot fiber, defatted linseed flour, and combinations thereof.

[0257] In some embodiments, the flavoring can be black salt, sea salt, onion powder, and combinations thereof. In some embodiments, the flavoring can be black salt, black pepper, Himalayan sea salt, salt, onion powder, minced onion, roasted garlic, mushroom powder, yeast extract, and combinations thereof. In some embodiments, the seasoning can be selected from almond extract, anise, basil, bay leaves, BBQ seasoning (various), black garlic powder, brown sugar, cajun seasoning, caraway, cardamom, cayenne pepper, celery salt, chili, chipotle, chives, cinnamon, cloves , cocoa powder, cumin, dill, fennel, five spice, garam masala, garlic (various), ginger, jalapeno, lavender, lemon, lime, maple sugar, maqoram, molasses, mulling spices, mustard seed, mustard powder, nutmeg, onion powder, oregano, paprika pepper (various), parsley, peppermint extract, poppy seeds, poultry seasoning, red chili flakes, rosemary, saffron, sage, salt (various), sesame seeds, soy, star anise, sugar, tarragon, thyme, tumeric, vanilla, wasabi, and combinations thereof.

[0258] In some embodiments of the dairy analogue composition described herein, each of the one or more additional ingredient is in an amount of about 0.05% to about 5%, about 0.05% to about 4%, or about 0.05% to about 3%, or about .05% to about 2%. In certain embodiments of the dairy analogue composition described herein, each of the one or more additional ingredient is in an amount of about 0.1% to about 1%.

[0259] In some embodiments of the dairy analogue composition described herein, the dairy analogue composition further comprises one or more hydrocolloids. In some embodiments the one or more hydrocolloids can be locust bean gum, a guar gum, a konjac gum, a gellan gum, a high methoxy pectin, a low methoxy pectin, an Agar, a kappa carrageenan, an iota carrageenan, a lambda carrageenan, an alginate, a curdlan, a methyl cellulose, a carboxymethyl cellulose (CMC), a xanthan gum, a gum Arabic, a Euglena derived beta-glucan and combinations thereof. [0260] In some embodiments of the dairy analogue composition described herein, each of the one or more hydrocolloids is in an amount of about 0.05% to about 8%, about 0.1% to about 8%, about 0.05% to about 7%, about 0.05% to about 6%, about 0.05% to about 5%, about 0.05% to about 4%, or about 0.05% to about 3%. In certain embodiments of the dairy analogue composition described herein, each of the one or more hydrocolloids is in an amount of about 0.1% to about 2%.

[0261] In some embodiments of the dairy analogue composition described herein has a similar nutritional content as a natural animal dairy product, which previous attempts to create a meat substitute have failed to attain.

[0262] In some embodiments described herein, the dairy analogue or food composition provides the same functional benefits as a natural animal dairy product. The functional properties are measured and evaluated using the dairy analogue or food product (e.g., yogurt, creamer, cheese sauce, butter, cream cheese, cheese, etc.) described herein. In some embodiments, taste can be measured using chromatographic analysis and consumer testing. In some embodiments, color can be measured using a colorimetric analyzer. The dairy analogue composition or food product described herein has been developed to provide some or all of the same functional benefits or properties as a natural dairy product. Functional properties can include, for example, water absorption and retention, solubility, color, gelation, viscosity, texture, emulsification properties, foam formation, flavor-binding properties, curdling, enzymatic browning, dextrinisation, caramelisation, flavour, preserving properties (e.g., shelf life), gelation, denaturation, coagulation, gluten formation, shortening, plasticity, aeration, flakiness, retention of moisture, sensory attributes, taste, and combinations thereof. That is, a dairy analogue or food product as described herein can have one or more functional properties that are the same or similar when compared to a natural dairy product such as natural milk from a cow, sheep, goat, buffalo, etc. or yogurt, creamer, cheese sauce, butter, cream cheese, or cheese made from these natural milks. For example, a yogurt made with a dairy analog as described herein can have the same or similar gelation, taste, and color of a yogurt made with natural cow’s milk.

[0263] In some embodiments described herein, the dairy analogue composition has a shelf life which is longer than a natural animal dairy product at either room temperature, refrigerator temperature (about 2°C), or freezer temperature (about -4°C). In certain embodiments, the dairy analogue composition contains no added preservatives. The dairy analogue composition described herein has an enhanced shelf life at room temperature. In some embodiments, the shelf life at room temperature is greater than about 3 months, greater than about 4 months, greater than about 5 months, greater than about 6 months, greater than about 7 months, greater than about 8 months, greater than about 9 months, greater than about 10 months, greater than about 11 months, or greater than about 12 months. In some embodiments, the shelf life at room temperature is at least about 6 months. The dairy analogue composition described herein has an enhanced shelf life when stored in a refrigerator or freezer. In some embodiments, the shelf life when stored in a refrigerator or freezer is greater than about 3 months, greater than about 4 months, greater than about 5 months, greater than about 6 months, greater than about 7 months, greater than about 8 months, greater than about 9 months, greater than about 10 months, greater than about 11 months, or greater than about 12 months. In some embodiments, the shelf life when stored in a refrigerator or freezer is at least about 6 months. In some embodiments, the dairy analogue composition described herein has the freeze stability and thaw stability of a natural animal dairy product.

[0264] Some embodiments describe a food product comprising a dairy analogue composition according to any embodiment described herein. Some embodiments described herein are directed to a food product comprising a dairy analogue composition comprising about 1% to about 100% (e.g., about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena- derived material and one or more additional ingredient, wherein the dairy analogue composition comprises one or more functional properties of a natural animal dairy product.

[0265] In some embodiments described herein, the one or more additional ingredients are selected from a protein, dietary fiber, a fat, a sugar, a sensory peptide or amino acid, or combinations thereof.

[0266] In some embodiments described herein, the protein is selected from Euglena flour, soy flour, chickpea flour, lentils flour, almond flour, legume flour, quinoa flour, millet flour, sorghum flour, or any other reasonable flour, protein concentrate or protein isolate source, and combinations thereof.

[0267] In some embodiments, the dairy analogue can include about 0.001% to about 30% yeast, live yeast, probiotics, whole cell inactive yeasts, yeast fractions, yeast extracts, yeast cell walls, yeast flakes. In some embodiments, the dairy analogue can include optionally about 0.05% to about 15% yeast, live yeast, probiotics, whole cell inactive yeasts, yeast fractions, yeast extracts, yeast cell walls, yeast flakes.

[0268] In some embodiments, the dairy analogue can include about 0.001% to about 30% nutritional yeast. In some embodiments the dairy analogue can include optionally about 0.05% to about 15% nutritional yeast. [0269] In some embodiments described herein, the dietary fiber is selected from cellulose, inulin and oligofructose, pectins, beta glucans, psyllium, lignin, resistant starch, hemicelluloses, gums, chitin and chitosan, fructooligosaccharides, galactooligosaccharides, poly dextrose and polyols, resistant dextrins, glucomannan, CitriFi® 100 FG (natural citrus fiber), methylcellulose, and combinations thereof.

[0270] In some embodiments described herein, the fat is selected from vegetable or seed oils derived from plants, or microbial source including without limitation, an oil derived from soy, rapeseed, flaxseed, walnut, canola, palm, soybean, palm kernel, coconut, com, olive, sunflower, cotton seed, cuphea, peanut, cashew, camelina sativa, mustard seed, cashew nut, oats, lupine, kenaf, calendula, hemp, coffee, linseed, hazelnut, euphorbia, pumpkin seed, coriander, camellia, sesame, safflower, rice, tung oil tree, cocoa, copra, opium poppy, castor beans, pecan, jojoba, jatropha, macadamia, Brazil nuts, and avocado, Euglena derived oil, medium-chain triglycerides (MCT), omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and oleic acid, and combinations thereof. In some embodiments, the fat is butter, lard, fats from fatty fish (e.g., salmon tuna, mackerel, herring, trout, sardines, or other suitable fish), soymilk derived fat, tofu derived fat, omega 3s, olive oil, sesame oil, avocado fat, olives, peanut butter, or combinations thereof.

[0271] In some embodiments, the dairy analogue can include about 0.1% to about 50% fat. In some embodiments, the dairy analogue can include one oil or a combination of oils, comprising about 0.1% to about 30% fat.

[0272] In some embodiments described herein, the dietary sugar is selected from granulated sugar, brown sugar, dark brown sugar, golden brown sugar, raw sugar, brown rice syrup, rice syrup, coconut sugar, com syrup, high-fructose com symp, maple symp, honey, agave, agave nectar, agave symp, cane sugar, dextrose, glucose, sucrose, fructose, galactose, lactose, maltose, molasses, com steep liquids, turbinado sugar, yellow sugar, muscovado sugar, maltodextrin, icing sugar, grape sugar, date sugar, confectioner’s sugar, beet sugar, and combinations thereof.

[0273] Sensory peptides and amino acids are compounds that influence the taste of foods. A database of sensory peptides and amino acids can be found at, for example, Iwaniak etal., BIOPEP database of sensory peptides and amino acids, Food Res Int 2016 Jul;85: 155- 161, which is incorporated herein by reference. In some embodiments described herein, a sensory peptide or amino acid is selected from bitter peptides, manchego cheese peptides, sour amino acids, sour peptides, umami amino acids, umami peptides, salty amino acids, astringent amino acids, sweet peptides, sweet amino acids, delicious peptide (umami), delicious peptide (sour), sourness suppressing peptide, sweetness suppressing peptide, umami enhancing peptide, salty taste enhancing peptide, kokumi peptide, and combinations thereof.

[0274] In some embodiments described herein, the food product comprises a preservative. In some embodiments, the food product has no added preservative. The preservative can be selected from Refresh 386, acetic acid, ascorbic acid, calcium ascorbate, erythorbic acid, iso-ascorbic acid, potassium nitrate, potassium nitrite, sodium ascorbate, sodium erythorbate, sodium iso-ascorbate, sodium nitrate, sodium nitrite, wood smoke, benzoic acid, calcium sorbate, Carnobacterium divergens M35, Carnobacterium maltaromaticum CB1, ethyl lauroyl arginate, 4-Hexyresoricinol, Leuconostoc carnosum 4010, methyl -/-hydroxybenzoate (methylparaben), modified vinegar (a liquid or spray-dried mixture containing acetic acid and one or more of potassium acetate, potassium diacetate, sodium acetate or sodium diacetate, prepared by adding potassium bicarbonate or sodium bicarbonate, potassium carbonate or sodium carbonate or potassium hydroxide or sodium hydroxide to vinegar), nisin, potassium acetate, potassium, benzoate, potassium bisulphite, potassium diacetate, potassium lactate, potassium, metabisulphite, potassium sorbate, propionic acid, propyl-/ -hydroxybenzoate (propylparaben), sodium acetate, sodium benzoate, sodium bisulphite, sodium diacetate, sodium lactate, sodium metabisulphite, sodium propionate, sodium salt of methyl-p-hydroxybenzoate acid, sodium salt of propyl-/ -hydroxybenzoate. sodium sorbate, sodium sulphite, sodium dithionite, sorbic acid, sulphurous acid, calcium propionate, calcium sorbate, dimethyl decarbonate, natamycin, potassium sorbate, propionic acid, sodium diacetate, sodium propionate, sodium sorbate, sorbic acid, asorbic acid, ascorbyl palmitate, ascorbyl stearate, butylated hydro-xyanisole (a mixture of 2-tertiarybutyl-4- hydroxyanisole and 3-tertiarybutyl-4- hydroxyanisole), butylated hydroxytoluene (3,5- ditertiarybutyl-4-hydroxytoluene), citric acid, citric acid esters of mono- and diglycerides, L- cysteine, L-cysteine hydrochloride, gum, guaiacum, lecithin, lethicin citrate, monoglyceride citrate, monoisopropyl citrate, propyl, gallate, sodium metabisulphite, tartaric acid, tertiary butyl hydroquinone, tocopherols (alpha-tocopherol; tocopherols concentrate, mixed), nitrites (nitrates and nitrosamines), and combinations thereof. In some embodiments the dairy analogue can include about 0.05% to about 10% preservative. In some embodiments, the dairy analogue can include optionally about 0.1% to about 2% preservative.

[0275] In some embodiments described herein, the food product comprises an antioxidant. The antioxidant can be selected from Dadex ARHS, ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), flavonoids, tannins, phenols, lignans, ascorbic acid, glutathione, melatonin, tocopherols, tocotrienols, uric acid, beta- carotene, lycopene, lutein, lipoic acid, vitamin E, selenium, manganese, and combinations thereof. In some embodiments, the dairy analogue can include about 0.0001% to about 10% antioxidants. In some embodiments, the dairy analogue can include optionally about 0.0001% to about 2% antioxidants.

|0276] In some embodiments described herein, the food product comprises a yeast. The yeast can be, active dry (traditional) yeast, instant yeast, bread machine/pizza yeast, rapid rise (or quick rising) instant yeast, fresh yeast, live yeast/probiotics, whole cell inactive yeasts, yeast fractions, yeast extracts, yeast cell walls, nutritional yeast, and combinations thereof.

[0277] In some embodiments described herein, the food product is selected from yogurt, butter, creamer, cream cheese, cheese sauce, cheese food products, sour cream, cheese curds, shredded cheese, block cheese, cheese based product, table cream, whipping cream, cheese strings, cottage cheese, flavoured cheese, a milk, a buttermilk, and combinations thereof.

[0278] Embodiments described herein are directed to food product comprising a dairy analogue composition comprising about 1% to about 100% (e.g., about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena- derived material, a Euglena beta glucan isolate emulsion, and one or more additional ingredients, wherein the dairy analogue food product comprises one or more functional properties of a natural dairy product.

[0279] In some embodiments, the Euglena beta glucan isolate is more than about 10% of the dairy analogue.

[0280] In some embodiments, the Euglena beta glucan isolate emulsion has an emulsion micelle size of about 10 mm to about 200 mm.

[0281] In some embodiments, the Euglena beta glucan isolate comprises a gelation agent in an amount of about 0% to about 1%.

[0282] In some embodiments, a dairy analogue composition comprising about 1 to about 10% Euglena-dcnvcd material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

Method of Making a Dairy Analogue

[0283] In some embodiments of the dairy analogue composition herein, methods of making and/or producing the plant-based protein product of the present disclosure comprise processes including, but not limited to preconditioning (e.g., to hydrate a dry mix), mixing, conveying, extruding, compressing, cooking, heating, shearing, cooling, tenderizing, marinating, and/or drying. [0284] The plant-based protein product of the present disclosure may be made and/or produced by introducing one or more protein sources, a plurality of dry ingredients, and a medium, along with any optional enhancer components (e.g., a flavoring agent) to an apparatus, an instrument, or an equipment, such as an extruding apparatus

|0285] In some embodiments, methods of making a yogurt are provided. A milk alternative, lecithin, water, Euglena- derived material, sugar, and flour can be mixed over heat to combine. In an embodiment the milk alternative can be, for example, cocoa butter powder, or nut, coconut, oat milk or powder. The milk alternative can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The lecithin can be, for example, sunflower lecithin. The lecithin can be in an amount of about 0.01% to about 50% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50%). The Euglena- derived material can be, for example, Euglena beta-glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The sugar can be, for example cane sugar. The sugar can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The flour can be, for example, oat flour. The flour can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The mixture can be heated to a temperature of about 160 degrees Fahrenheit to about 190 degrees Fahrenheit for 5, 10, 15, 20, 30, 60, 120 minutes or longer. The mixture can be cooled to about 80 to 140 degrees Fahrenheit while stirring. A suitable acid such as lactic acid can be added to the mixture. The suitable acid can be in an amount of about 0,01% to about 50%, about 0.01% to about 20%, or about 0.01% to about 5% (e.g. about 0.01, 1, 5, 10, 20, 30, 40, or 50%).

[0286] The mixture can be set at about 35 to about 50 degrees Fahrenheit for about 12, 24, 36 hours or more. The yogurt has one or more functional properties of a yogurt made with a natural dairy source (e.g., cow’s milk). A yogurt bacterial culture can be any suitable bacterial culture used to make yogurt such as Lactobacillus bulgaricus, Lactobacillus delbrueckii, Streptococcus thermophilus, and combinations thereof.

[0287] In some embodiments, the method of making a dairy analogue, wherein a yogurt bacterial culture such as a frozen yogurt bacterial culture, is added prior to the step of setting the mixture.

[0288] In some embodiments, the method of making a dairy analogue yogurt formulation comprising of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena- derived material, about 0.05%to about 70% additional liquid source and one or more additional ingredient, wherein the yogurt formulation comprises of one or more properties of natural animal yogurt.

[0289] In some embodiments, the method of making a dairy analogue yogurt formulation, wherein the Euglena- derived material was a dry material. In some embodiments of the yogurt formulation described herein, the Euglena- derived material was a wet material. In some embodiments the yogurt formulation described herein used both wet and dry Euglena derived material.

[0290] In some embodiments, methods of making a butter are provided. An oil, Euglena- derived material, and yeast can be mixed over heat to combine. In an embodiment, the oil can be, for example coconut oil. The oil can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the Euglena- derived material can be, for example, Euglena beta-glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the yeast can be, for example, nutritional yeast. The yeast can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The mixture can be heated to a temperature of about 100 degrees Fahrenheit to about 190 degrees Fahrenheit for 5, 10, 15, 20, 30, 60, 120 minutes or longer. The mixture can be cooled to about 30 degrees Fahrenheit to about 140 degrees Fahrenheit.

[0291] In some embodiments, methods of making a butter are provided. The method comprising heating oil; adding Euglena flour and yeast extract to the oil while stirring to make a mixture; cooling the mixture; and blending the mixture at least one time; and allowing the mixture to solidify; to create a dairy analogue comprising one or more functional properties of a natural butter.

[0292] In some embodiments, the method of making a dairy analogue, wherein the Euglena flour is a refined flour. In some embodiments, the Euglena flour further comprises a masker and an antioxidant.

[0293] In some embodiments, the method of making a butter formulation comprising of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena- derived material, about 0.05% to about 70% additional liquid source and one or more additional ingredients, wherein the butter formulation comprises of one or more properties of a natural animal butter.

[0294] In some embodiments of the butter formulation described herein, the Euglena-dcnvcd material was a dry material. In some embodiments of the butter formulation described herein, the Euglena- derived material was a wet material. In some embodiments the butter formulation described herein used both wet and dry Euglena derived material.

[0295] In some embodiments, the method of making a butter, wherein the mixture comprises a Euglena beta glucan isolate. In some embodiments, the method of making a dairy analogue, wherein the mixture comprises a flavourant.

[0296] In some embodiments, methods of making a creamer are provided. A Euglena-dcnvcd material, oil, lecithin, a protein source, and an emulsifier are combined. In an embodiment, the oil can be, for example, coconut oil. The oil can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The lecithin can be, for example, sunflower lecithin. The lecithin can be in an amount of about 0.01% to about 50% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50%). In an embodiment, the Euglena- derived material can be, for example, Euglena beta-glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the protein source can be, for example pea protein. The protein source can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the emulsifier can be, for example, Myverol 19-02K. The emulsifier can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). Embodiments described herein a directed to a method of making a dairy analogue, wherein the dairy analogue is a creamer, the method comprising mixing Euglena- derived material, oil, lecithin, a protein source, and an emulsifier to create a dairy analogue comprising one or more functional properties of a natural creamer.

[0297] In some embodiments, the method of making a creamer, wherein the Euglena- derived material is a ready to gel Euglena beta glucan isolate, a wet Euglena beta glucan isolate, a Euglena beta glucan slurry, a Euglena protein concentrate, a Euglena protein isolate, a Euglena oil, a Euglena biomass, a Euglena flour, a Euglena milled paramylon, an emulsified Euglena beta glucan isolate, and combinations thereof.

[0298] In some embodiments, methods of making a cream cheese are provided. A Euglena- derived material, a protein source, a hydrocolloid, and a lecithin can be combined and hydrated to create a beta glucan isolate slurry. The hydrated mixture can then be emulsified and cooled. In an embodiment, the Euglena- derived material can be, for example, Euglena beta-glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The protein source can be, for example, pea protein. The protein source can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the hydrocolloid may be, for example, xantham gum. The hydrocolloid source can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In some embodiments, the lecithin can be, for example, sunflower lecithin. The lecithin can be in an amount of about 0.01% to about 50% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50%). Embodiments described herein are directed to a method of making a dairy analogue, wherein the dairy analogue is a cream cheese, the method comprising creating a beta glucan isolate slurry comprising Euglena beta glucan isolate, a protein source, a hydrocolloid, and lecithin to form a mixture; hydrating the mixture for about 1 hour to about 24 hours; emulsifying an oil to form an emulsified oil; combining the emulsified oil to the mixture while stirring; cooling the emulsified oil and the mixture combination for about 12 to about 16 hours; and blending a suitable acid into the oil and the mixture. The mixture can be cooled to about 35 to about 50 degrees Fahrenheit. The suitable acid can be, for example, citric acid.

[0299] In some embodiments, the method of making a cream cheese wherein the mixture is hydrating for about 1 to about 6 hours. In some embodiments, the method of making a cream cheese wherein the mixture is hydrating for at least about 2 hours.

[0300] In some embodiments, the method of making a dairy analogue cream cheese comprising about 0.1% to about 100% (e.g., about 0.1,1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena- derived material, about 0.05% to about 70% additional liquid source and one or more additional ingredient, wherein the cream cheese comprises of one or more properties of natural animal cream cheese.

[0301] In some embodiments, the method of making a dairy analogue cream cheese, wherein the Euglena-dcnvcd material is a dry material. In some embodiments, the Euglena- derived material is a wet material. In some embodiments the method comprises both wet and dry Euglena-dcmcd material.

[0302] In some embodiments, methods of making a cream cheese are provided. A Euglena-dcm cd material, a protein source, a hydrocolloid, a lecithin, an oil, and a gelling agent can be combined. In an embodiment, the Euglena-dcmcd material can be, for example, Euglena beta-glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The protein source can be, for example, pea protein. The protein source can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the hydrocolloid may be, for example, xantham gum. The hydrocolloid source can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In some embodiments, the lecithin can be, for example, sunflower lecithin. The lecithin can be in an amount of about 0.01%to about 50% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50%).In some embodiments, the oil can be, for example, coconut oil. The oil can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In some embodiment, the gelling agent can be, for example, Ready to Gel. The gelling agent can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). Embodiments described herein are directed to a method of making a dairy analogue, wherein the dairy analogue is a cream cheese, the method comprising mixing a protein source, one or more hydrocolloids, xanthan and water for form a mixture; adding a suitable acid to the mixture, wherein the mixture has a pH of about pH 3.5 to about pH 4.5; stirring the mixture; adding to the mixture a gelation agent, oil, and lecithin.

[0303] In some embodiments, the method of making a cream cheese wherein the cream cheese comprises a flavourant.

[0304] In some embodiments, method of making a cheese sauce are provided. A milk alternative, a Euglena- derived material, a yeast, a protein source, an oil, a starch, and a hydrocolloid are combined. In an embodiment the milk alternative can be, for example, cocoa butter powder, or nut, coconut, oat milk or powder. The milk alternative can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The Euglena- derived material can be, for example, Euglena beta- glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The yeast can be, for example nutritional yeast. The yeast can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The protein source can be, for example pea protein. The protein source can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The oil can be, for example, coconut oil. The oil can be in an amount of about 0.1% to about 100% (e.g., about 0.1 _, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The starch can be, for example oat starch or pea starch. The starch can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The hydrocolloid can be, for example, xantham gum. The hydrocolloid can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The method comprises mixing a milk alternative, a Euglena source, nutritional yeast, a protein source, a coconut oil, a starch source, and a hydrocolloid, to create a mixture; and blending the mixture.

|0305] In some embodiments, method of making a cheese are provided. A milk alternative, a Euglena- derived material, an oil, an antibacterial agent, an antifungal agent, a hydrocolloid, and optionally a yeast are combined then heated followed by cooling. In an embodiment the milk alternative can be, for example, cocoa butter powder, or nut, coconut, oat milk or powder. The milk alternative can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). Th e Euglena- derived material can be, for example, Euglena beta-glucan isolate. The Euglena- derived material can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The yeast can be, for example, nutritional yeast. The yeast can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The oil can be, for example, coconut oil. The oil can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The hydrocolloid can be, for example, xantham gum. The hydrocolloid can be in an amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The antibacterial agent can be acetic acid, benzoic acid, acetic acid, nisin, nitrates, nitrites, propionic acid, sorbic acid, sulfites and sulfur dioxide. The antibacterial agent can be in an amount of about 0.01% to about 50% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50%). The antifungal agent can be acetic acid, benzoic acid, natamycin, propionic acid, sorbic acid, sultites and sulfur dioxide, parabens, ethyl formate. The antifungal agent can be in an amount of about about 0.01% to about 50% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50%). Embodiments described herein are directed to a method of making a dairy analogue, wherein the dairy analogue is a cheese, the method comprising combining a hydrocolloid, Euglena protein concentrate, a suitable anti-bacterial agent, and a suitable anti-fungal agent to create a mixture; adding oil and liquid to the mixture; heating to about 160 degrees Fahrenheit to about 190 degrees Fahrenheit; and storing at about 35 degrees Fahrenheit to about 50 degrees Fahrenheit for at least about 7 days. The mixture can be heated to a temperature of about 100 degrees Fahrenheit to about 190 degrees Fahrenheit for 5, 10, 15, 20, 30, 60, 120 minutes or longer. The mixture can be cooled to about 30 degrees Fahrenheit to about 140 degrees Fahrenheit. [03061 In some embodiments, the method of making a cheese wherein the mixture further comprises a yeast extract.

[0307] In some embodiments, the method of making a cheese comprising of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena-dcnvcd material, about 0.05% to about 70% additional liquid source and one or more additional ingredient, wherein the cheese formulation comprises of one or more properties of natural animal cheese.

[0308] In some embodiments, the method of making a cheese comprising Euglena- derived material, wherein the Euglena- derived material is a dry material. In some embodiments the Euglena- derived material was a wet material. In some embodiments the method comprises both wet and dry Euglena derived material.

[0309] Embodiments described herein are directed to a method of making a Euglena beta glucan isolate emulsion, the method comprising incubating Euglena beta glucan isolate in oil for about 12 to about 18 hours; adding water to the beta glucan isolate and oil to create a mixture; and homogenizing the mixture.

[0310] In some embodiments, the method of making a Euglena beta glucan isolate emulsion wherein the oil is selected from palm oil, canola oil, linseed oil, sunflower oil, coconut oil, or combinations thereof.

[0311] In some embodiments, the method of making a Euglena beta glucan isolate emulsion wherein the Euglena beta glucan isolate emulsion has an emulsion micelle size of about 10 mm to about 200 mm. In some embodiments, the emulsion micelle size is about 50 mm to about 100 mm.

[0312] Embodiments described herein are directed to a method of making a ready to gel beta glucan isolate, the method comprising: 1) dissolving beta glucan isolate in 1M NaOH to form a 5% paramylon - NaOH solution; 2) adding 35% citric acid solution to the 5% paramylon - NaOH solution; 3) stirring the solution at increasing speeds until the solution is mixing vigorously, wherein a gel will begin to form at about pH 6 or higher, and wherein a 35% citric acid solution can be used to adjust the pH of the solution; 4) removing the stir bar after the gel is formed, wherein the gel can be used as a wet gel product or formed into a ready to gel powder; 5) blending the gel with a blender; 6) centrifuging the gel for about 10 to about 20 minutes at about 3500 rpm; 7) pouring off the supernatant and adding deionized water; 8) mixing the gel and water vigorously for 1 minute, wherein no clumps exist after stirring; 9) centrifuging the gel at 3500 rpm for about 10 minutes; 10) decanting the supernatant and adding deionized water to wash the gel; 11) repeating the washing about six times; 12) drying the gel 13) freezing the gel at about -80 degrees Celsius; 14) drying the frozen gel in the freezer dryer; 15) grinding the freezer dried gel into a fine powder to form a dry gel; 16) optionally forming a wet gel by adding water to the dry gel to form a final ready to gel powder concentration of at least about 5% or at least about 10% Ready To Gel powder in water.

Meat Analogue Compositions

[0313] Embodiments described herein are directed to a meat analogue composition comprising about 0.5% to about 95% W/W Euglena- derived material, and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural animal meat product.

[0314] In some embodiments described herein, the meat analogue composition comprises about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena- derived material, about 0.05% to about 70% additional protein source, and one or more additional ingredient. In some embodiments, the meat analogue composition comprises about 10% to about 98% Euglena- derived material. In some embodiments, the meat analogue composition comprises about 0.01% to about 100% Euglena- derived material. In some embodiments, the meat analogue composition comprises about 0.01%, about 0.05%, about 1%, about 2%, about 3%, about 4% about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100% Euglena- derived material or any range within these values.

[0315] In some embodiments described herein, the Euglena-dcmcd material can be Euglena biomass (e.g., fresh, wet or dry biomass), a Euglena derived protein (i.e., Euglena protein, e.g. wet or dry), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate (e.g. wet or dry), a Euglena protein isolate (e.g. wet or dry), a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, Ready To Gel Euglena beta-glucan isolate, Euglena beta-glucan slurry, a Euglena oil, milled Euglena paramylon (e.g. wet or dry), Euglena wet protein concentrate, Euglena dry protein concentrate, and combinations thereof.

[0316] In some embodiments, a meat analogue composition comprising about 10 to about 20% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product. [0317] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0318] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0319] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0320] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0321] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0322] In some embodiments, a meat analogue composition comprising about 20 to about 30% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0323] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0324] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0325] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0326] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0327] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0328] In some embodiments, a meat analogue composition comprising about 30 to about 40% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0329] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0330] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0331] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0332] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0333] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [0334] In some embodiments, a meat analogue composition comprising about 40 to about 50% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

|0335] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0336] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0337] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0338] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0339] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0340] In some embodiments, a meat analogue composition comprising about 50 to about 60% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0341] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0342] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1%to about 99.9% W/W ofthe composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0343] hi some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0344] hi some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0345] hi some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0346] hi some embodiments, a meat analogue composition comprising about 60 to about 70% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0347] hi some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0348] hi some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0349] hi some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0350] hi some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [03511 In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0352] In some embodiments, a dairy analogue composition comprising about 70 to about 80% W/W Euglena- derived material and one or more additional ingredients, wherein the dairy analogue composition comprises one or more functional properties of a natural dairy product.

[0353] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0354] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0355] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0356] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0357] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0358] In some embodiments, a meat analogue composition comprising about 80 to about 90% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0359] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0360] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0361] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0362] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0363] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0364] In some embodiments, a meat analogue composition comprising about 90 to about 99.5% W/W hug/ena-dcmcd material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0365] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0366] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0367] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [03681 In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0369] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0370] In some embodiments, a meat analogue composition comprising about 5 to about 80% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0371] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0372] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0373] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0374] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0375] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0376] In some embodiments, a meat analogue composition comprising about 10 to about 90% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0377] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0378] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1% to about 99.9% W/W of the composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0379] In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0380] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0381] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0382] In some embodiments, a meat analogue composition comprising about 20 to about 50% W/W Euglena- derived material and one or more additional ingredients, wherein the meat analogue composition comprises one or more functional properties of a natural meat product.

[0383] In some embodiments, the one or more additional ingredients is a protein source, wherein the protein source is about 0.1% to about 99.9% W/W of the composition. The protein source can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0384] In some embodiments, the one or more additional ingredients is a hydrocolloid, wherein the hydrocolloid is about 0.1%to about 99.9% W/W ofthe composition. The hydrocolloid can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%). [03851 In some embodiments, the one or more additional ingredients is a fat, wherein the fat is about 0.1% to about 99.9% W/W of the composition. The fat can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

|0386] In some embodiments, the one or more additional ingredients is a flavouring, wherein the flavouring is about 0.1% to about 99.9% W/W of the composition. The flavouring can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0387] In some embodiments, the one or more additional ingredients is a sugar, wherein the sugar is about 0.1% to about 99.9% W/W of the composition. The sugar can be any amount of about 0.1% to about 100% (e.g., about 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9%).

[0388] In some embodiments of the meat analogue described herein, the fresh Euglena- derived material can be Euglena biomass (e.g., fresh, wet or dry biomass), a Euglena derived protein (i.e., Euglena protein), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate, a. Euglena protein isolate, a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, ready to gel Euglena beta-glucan isolate, Euglena beta-glucan slurry, a Euglena oil, milled Euglena paramylon, Euglena wet protein concentrate, and combinations thereof. Euglenid protozoans (e.g., Euglena gracilis) are a source of an insoluble, linear (l,3)- -glucan of high molecular mass called paramylon that occurs naturally in a high crystalline form in discrete membrane- bound granules in the cytoplasm.

[0389] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena protein concentrate. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, optionally about 3% to about 50%, or optionally about 0.5% to about 15% Euglena protein concentrate.

[0390] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena protein flour or wet biomass. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, optionally about 0.1% to about 60%, or optionally about 5% to about 30% Euglena protein flour or wet biomass.

[0391] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena protein isolate. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena protein isolate.

[0392] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena beta glucan isolate. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena beta glucan isolate.

[0393] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena textured vegetable protein. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena textured vegetable protein. In some embodiments, the meat analogue can include optionally about 5 to about 50% Euglena textured vegetable protein.

[0394] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena high moisture extrusion product. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena high moisture extrusion product.

[0395] In some embodiments, the meat analogue can include about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%) Euglena low moisture extrusion product. In some embodiments, the meat analogue can include optionally about 0.1% to about 75%, or optionally about 1% to about 50% Euglena low moisture extrusion product.

[0396] In some embodiments, the Euglena- derived wet protein concentrate is in an amount of about 5% to about 95%, about 15% to about 95%, about 20% to about 95%, about 25% to about 95%, about 30% to about 95%, about 30% to about 95%, about 35% to about 95%, about 40% to about 95%, about 45% to about 95%, about 50% to about 95%, about 55% to about 95% W/W, in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 94% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 56% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 7% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 40% to about 87% in a liquid mixture. In certain embodiments, the Euglena- derived wet protein concentrate is in an amount of about 40% to about 90% in a liquid mixture.

[0397] In some embodiments, the Euglena-AcmcA wet protein concentrate is about 25% to about 80% protein, about 30% to about 80% protein, about 35% to about 80% protein, about 40% to about 80% protein, about 45% to about 80% protein, about 50% to about 80% protein, about 55% to about 80% protein, about 60% to about 80% protein, about 65% to about 80% protein, or about 25%to about 75% protein. In certain embodiments, the Euglena- derived wet protein concentrate is about 70% protein.

[0398] In some embodiments, the Euglena- derived wet protein concentrate is an Euglena- derived protein isolate.

[0399] In some embodiments, the Euglena protein concentrate can be about 5% to about 50% ofthe composition. The amount of Euglena protein concentrate can vary depending on the type of composition. In some embodiments, the Euglena protein concentrate can be wet Euglena protein concentrate. In some embodiments, the Euglena protein concentrate can be dry Euglena protein concentrate. In some embodiments there the ratio of moisture content to protein is 2: 1. In some embodiments, the moisture content of protein concentrate is about 50% to about 90%. In some embodiments, the moisture content of protein concentrate is about 70 to about 90%.

[0400] In some embodiments, the protein concentrate is a ready to gel beta glucan isolate, wherein the protein concentrate is in an amount of about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In certain embodiments, the protein concentrate is a ready to gel beta glucan isolate, wherein the protein concentrate is in an amount of about 0.1% to about 20%.

[0401] In some embodiments, the protein concentrate is a beta glucan slurry or a milled paramylon, wherein the protein concentrate is in an amount of about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In some embodiments, the protein concentrate is a beta glucan slurry or a milled paramylon, wherein the protein concentrate is in an amount of about 0.1% to about 100 %. In some embodiments, the protein concentrate is a beta glucan slurry or a milled paramylon, wherein the protein concentrate is in an amount of about 5% to about 95% W/W.

[0402] In some embodiments, the Euglena- derived dry protein concentrate is in an amount of about 5% to about 95%, about 15% to about 95%, about 20% to about 95%, about 25% to about 95%, about 30% to about 95%, about 30% to about 95%, about 35% to about 95%, about 40% to about 95%, about 45% to about 95%, about 50% to about 95%, about 55% to about 95% W/W, in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 94% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 56% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 7% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 87% in a liquid mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 90% in a liquid mixture.

[0403] In some embodiments, the Euglena- derived dry protein concentrate is in an amount of about 5% to about 95%, about 15% to about 95%, about 20% to about 95%, about 25% to about 95%, about 30% to about 95%, about 30% to about 95%, about 35% to about 95%, about 40% to about 95%, about 45% to about 95%, about 50% to about 95%, about 55% to about 95% W/W, in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 94% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 56% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 7% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 87% in a dry mixture. In certain embodiments, the Euglena- derived dry protein concentrate is in an amount of about 40% to about 90% in a dry mixture.

[0404] In some embodiments, the Euglena- derived dry protein concentrate is about 25% to about 80% protein, about 30% to about 80% protein, about 35% to about 80% protein, about 40% to about 80% protein, about 45% to about 80% protein, about 50% to about 80% protein, about 55% to about 80% protein, about 60% to about 80% protein, about 65% to about 80% protein, or about 25%to about 75% protein. In certain embodiments, the Euglena-dcnvcd wet protein concentrate is about 70% protein.

[0405] In some embodiments, the Euglena-dcnvcd dry protein concentrate is an Euglena- derived protein isolate.

[0406] For the meat analogue using fresh biomass or wet or powdered Euglena protein concentrate maskers are required to mask the marine flavours of Euglena. An ideal masker should completely mask the off notes (both aroma and flavour) coming from Euglena during cooking and in cooked products without introducing any new flavours or aroma. Flavours can also be used to improve the flavour. Ideal flavour should provide a flavour that is similar to cooked real animal meat. In certain embodiments, the maskers can be purchased from Firmenich, IFF, Givaudan, Symrise, Mane, Fona, Flavor producer, McCormick, edlong, T Hasegawa. Sensient Flavors, Robertet SA, Prova, Wild/ADM, Takasago, Synergy, and others.

[0407] In some embodiments of the meat analogue composition described herein, the Euglena- derived material is a dry material. In some embodiments of the meat analogue composition described herein, the Euglena- derived material is a wet material.

[0408] In some embodiments of the meat analogue composition described herein, the Euglena- derived material can be Euglena biomass (e.g., fresh, wet or dry biomass), a Euglena derived protein (i.e., Euglena protein), a Euglena flour (e.g., Euglena protein-rich flour), a Euglena protein concentrate, a Euglena protein isolate, a Euglena beta-glucan isolate (e.g., wet or dry), a Euglena beta-glucan rich flour, emulsified Euglena beta-glucan isolate, ready to gel Euglena beta-glucan isolate, Euglena beta-glucan slurry, a Euglena oil, milled Euglena paramylon, Euglena wet protein concentrate, and combinations thereof.

[0409] In some embodiments of the meat analogue composition described herein, the Euglena- derived material is in an amount of about 5% to about 90%, about 50% to about 85%, about 50% to about 75%, about 50% to about 65%, about 60% to about 85%, or about 70% to about 85% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 55% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 62% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 7% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 40% to about 87% in the mixture. In certain embodiments, the Euglena- derived material is in an amount of about 40% to about 90% in the mixture.

[0410] In some embodiments of the meat analogue composition described herein, the additional protein source can be pea protein, soy protein, com protein, wheat protein, rice protein, beans protein, seed protein, nut protein, almond protein, peanut protein, seitan protein, lentil protein, chickpea protein, flaxseed protein, wild rice protein , quom protein, chia seed protein, quinoa protein, oat protein, fava bean protein, buckwheat protein, bulgar protein, sorghum protein, millet protein, microalgae protein, yellow pea protein, mung bean protein, hemp protein, sunflower protein, canola protein, lupin protein, legumes protein, potato protein, and combinations thereof. The additional protein source may be any reasonable flour, protein concentrate, protein isolate source, and combinations thereof.

[0411] In some embodiments, the meat analogue can include about 0.1% to about 95% W/W pea protein. In some embodiments, the meat analogue can include optionally about 1% to about 95%, or optionally about 1% to about 75% W/W pea protein. [0412] In some embodiments of the meat analogue composition described herein, the additional protein source is in an amount of about 0.05% to about 70%, about 0.5% to about 70%, about l%to about 70%, about 5% to about 70%, about 10% to about 70%, about 15% to about 70%, about 0.05% to about 60%, about 0.05% to about 50%, about 0.05% to about 40%, about 0.05% to about 30%, or about 0.05% to about 20% W/W. In certain embodiments of the meat analogue composition described herein, the additional protein source is in an amount of about 0.05% to about 15% W/W. In certain embodiments of the meat analogue composition described herein, the additional protein source is in an amount of about 20% to about 60% WAV.

[0413] In some embodiments of the meat analogue composition described herein, the one or more additional ingredients are selected from a protein, a dietary fiber, a fat, a sugar, a sensory peptide or amino acid, or combinations thereof. In some embodiments of the meat analogue composition described herein, the one or more additional ingredient is can be of gellan gum, methylcellulose, yeast extract, flavoring, antioxidant blend, maskers, leavening agents, baking powder, baking soda, enzymes, transglutaminase, emulsifiers, lecithin, mono- and diglycerides, binders, carrot fiber, defatted linseed flour, and combinations thereof.

[0414] In some embodiments, the flavoring is can be of black salt, sea salt, onion powder, and combinations thereof. In some embodiments, the flavoring can be of black salt, black pepper, Himalayan sea salt, salt, onion powder, minced onion, roasted garlic, mushroom powder, yeast extract, and combinations thereof. In some embodiments, the seasoning can be selected from almond extract, anise, basil, bay leaves, BBQ seasoning (various), black garlic powder, brown sugar, cajun seasoning, caraway, cardamom, cayenne pepper, celery salt, chili, chipotle, chives, cinnamon, cloves , cocoa powder, cumin, dill, fennel, five spice, garam masala, garlic (various), ginger, jalapeno, lavender, lemon, lime, maple sugar, maqoram, molasses, mulling spices, mustard seed, mustard powder, nutmeg, onion powder, oregano, paprika pepper (various), parsley, peppermint extract, poppy seeds, poultry seasoning, red chili flakes, rosemary, saffron, sage, salt (various), sesame seeds, soy, star anise, sugar, tarragon, thyme, tumeric, vanilla, wasabi, and combinations thereof.

[0415] In some embodiments of the meat analogue composition described herein, each of the one or more additional ingredient is in an amount of about 0.05% to about 5%, about 0.05% to about 4%, or about 0.05% to about 3%, or about .05% to about 2%. In certain embodiments of the meat analogue composition described herein, each of the one or more additional ingredient is in an amount of about 0.1% to about 1%. [04161 In some embodiments of the meat analogue composition described herein, the meat analogue composition further comprises one or more hydrocolloids. In some embodiments the one or more hydrocolloids can be locust bean gum, a guar gum, a konjac gum, a gellan gum, a high methoxy pectin, a low methoxy pectin, an Agar, a kappa carrageenan, an iota carrageenan, a lambda carrageenan, an alginate, a curdlan, a methyl cellulose, a carboxymethyl cellulose (CMC), a xanthan gum, a gum Arabic, a Euglena derived beta-glucan and combinations thereof.

[0417] In some embodiments of the meat analogue composition described herein, each of the one or more hydrocolloids is in an amount of about 0.05% to about 8%, about 0.1% to about 8%, about 0.05% to about 7%, about 0.05% to about 6%, about 0.05% to about 5%, about 0.05% to about 4%, or about 0.05% to about 3%. In certain embodiments of the meat analogue composition described herein, each of the one or more hydrocolloids is in an amount of about 0.1% to about 2%.

[0418] In some embodiments of the meat analogue composition described herein has a similar nutritional content as a natural animal meat product, which previous attempts to create a meat substitute have failed to attain.

[0419] In some embodiments described herein, a meat analogue composition or food product provides the same functional benefits as a natural animal meat product. The functional properties are measured and evaluated using the meat analogue or food product (e.g., fish (e.g., flaked tuna or fish sticks), chicken, pulled pork, sausage, beef, ground beef) described herein. In some embodiments, taste can be measured using chromatographic analysis and consumer testing. In some embodiments, color can be measured using a colorimetric analyzer. The meat analogue composition or food product described herein has been developed to provide some or all of the same functional benefits or properties as a natural meat product. Functional properties can include, for example, water absorption and retention, solubility, color, gelation, viscosity, texture, emulsification, foam formation, flavor-binding properties, curdling, and enzymatic browning, dextrinisation, caramelisation, flavour, preserving properties (e.g., shelf life), gelation, denaturation, coagulation, gluten formation, shortening, plasticity, aeration, flakiness, retention of moisture, sensory attributes, taste, and combinations thereof. That is, a meat analogue or food product as described herein can have one or more functional properties that are the same or similar when compared to a natural meat product such as fish (e.g., flaked tuna or fish sticks), chicken, pulled pork, sausage, beef, ground beef, or products made from these natural meats. For example, pulled pork made with a meat analog as described herein can have the same or similar moisture retention, taste, texture, and color of pulled pork made from natural pork.

[0420] In some embodiments described herein, the meat analogue composition has a shelf life which is longer than a natural animal meat product at either room temperature, refrigerator temperature (about 2°C), or freezer temperature (about -4°C). In certain embodiments, the meat analogue composition contains no added preservatives. The meat analogue composition described herein has an enhanced shelf life at room temperature. In some embodiments, the shelf life at room temperature is greater than about 3 months, greater than about 4 months, greater than about 5 months, greater than about 6 months, greater than about 7 months, greater than about 8 months, greater than about 9 months, greater than about 10 months, greater than about 11 months, or greater than about 12 months. In some embodiments, the shelf life at room temperature is at least about 6 months. The meat analogue composition described herein has an enhanced shelf life when stored in a refrigerator or freezer. In some embodiments, the shelf life when stored in a refrigerator or freezer is greater than about 3 months, greater than about 4 months, greater than about 5 months, greater than about 6 months, greater than about 7 months, greater than about 8 months, greater than about 9 months, greater than about 10 months, greater than about 11 months, or greater than about 12 months. In some embodiments, the shelf life when stored in a refrigerator or freezer is at least about 6 months. In some embodiments, the meat analogue composition described herein has the freeze stability and thaw stability of a natural animal meat product.

[0421] Some embodiments describe a food product comprising a meat analogue composition according to any embodiment described herein. Some embodiments described herein are directed to a food product comprising a meat analogue composition comprising about 0.5% to about 95% W/W Euglena- derived material and one or more additional ingredient, wherein the meat analogue composition comprises one or more functional properties of a natural animal meat product.

[0422] In some embodiments described herein, the one or more additional ingredients are selected from a protein, dietary fiber, a fat, a sugar, a sensory peptide or amino acid, or combinations thereof.

[0423] In some embodiments described herein, the protein is selected from Euglena, soy, chickpea, lentils, almond, legume, quinoa, millet, sorghum, or any other reasonable flour, protein concentrate or protein isolate source, and combinations thereof.

[0424] In some embodiments, the meat analogue can include about 0.001% to about 30% yeast, live yeast, probiotics, whole cell inactive yeasts, yeast fractions, yeast extracts, yeast cell walls, yeast flakes. In some embodiments, the meat analogue can include optionally about 0.05% to about 15% yeast, live yeast, probiotics, whole cell inactive yeasts, yeast fractions, yeast extracts, yeast cell walls, yeast flakes.

[0425] In some embodiments, the meat analogue can include about 0.001% to about 30% nutritional yeast. In some embodiments the meat analogue can include optionally about 0.05% to about 15% nutritional yeast.

[0426] In some embodiments described herein, the dietary fiber is selected from cellulose, inulin and oligofructose, pectins, beta glucans, psyllium, lignin, resistant starch, hemicelluloses, gums, chitin and chitosan, fructooligosaccharides, galactooligosaccharides, poly dextrose and polyols, resistant dextrins, glucomannan, CitriFi® 100 FG (natural citrus fiber), methylcellulose, and combinations thereof.

[0427] In some embodiments described herein, the fat is selected from vegetable or seed oils derived from plants, or microbial source including without limitation, an oil derived from soy, rapeseed, flaxseed, walnut, canola, palm, soybean, palm kernel, coconut, com, olive, sunflower, cotton seed, cuphea, peanut, cashew, camelina sativa, mustard seed, cashew nut, oats, lupine, kenaf, calendula, hemp, coffee, linseed, hazelnut, euphorbia, pumpkin seed, coriander, camellia, sesame, safflower, rice, tung oil tree, cocoa, copra, opium poppy, castor beans, pecan, jojoba, jatropha, macadamia, Brazil nuts, and avocado, Euglena derived oil, medium-chain triglycerides (MCT), omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and oleic acid, and combinations thereof. In some embodiments. The fat is butter, lard, fats from fatty fish (e.g., salmon tuna, mackerel, herring, trout, sardines, or other suitable fish), soymilk derived fat, tofu derived fat, omega 3s, olive oil, sesame oil, avocado fat, olives, peanut butter, or combinations thereof.

[0428] In some embodiments, the meat analogue can include about 0.1% to about 50% fat. In some embodiments, the meat analogue can include one oil or a combination of oils, comprising about 0.1% to about 30% fat.

[0429] In some embodiments described herein, the dietary sugar is selected from granulated sugar, brown sugar, dark brown sugar, golden brown sugar, raw sugar, brown rice syrup, rice syrup, coconut sugar, com syrup, high-fructose com symp, maple symp, honey, agave, agave nectar, agave symp, cane sugar, dextrose, glucose, sucrose, fructose, galactose, lactose, maltose, molasses, com steep liquids, turbinado sugar, yellow sugar, muscovado sugar, maltodextrin, icing sugar, grape sugar, date sugar, confectioner’s sugar, beet sugar, and combinations thereof. [0430] In some embodiments described herein, asensory peptide or amino acid is selected from bitter peptides, manchego cheese peptides, sour amino acids, sour peptides, umami amino acids, umami peptides, salty amino acids, astringent amino acids, sweet peptides, sweet amino acids, delicious peptide (umami), delicious peptide (sour), sourness suppressing peptide, sweetness suppressing peptide, umami enhancing peptide, salty taste enhancing peptide, kokumi peptide, and combinations thereof.

[0431] In some embodiments described herein, the food product comprises a preservative. In some embodiments, the food product has no added preservative. The preservative can be selected from Refresh 386, acetic acid, ascorbic acid, calcium ascorbate, erythorbic acid, iso-ascorbic acid, potassium nitrate, potassium nitrite, sodium ascorbate, sodium erythorbate, sodium iso-ascorbate, sodium nitrate, sodium nitrite, wood smoke, benzoic acid, calcium sorbate, Carnobacterium divergens M35, Carnobacterium maltaromaticum CB1, ethyl lauroyl arginate, 4-Hexyresoricinol, Leuconostoc carnosum 4010, methyl -/-hydroxybenzoate (methylparaben), modified vinegar (a liquid or spray-dried mixture containing acetic acid and one or more of potassium acetate, potassium diacetate, sodium acetate or sodium diacetate, prepared by adding potassium bicarbonate or sodium bicarbonate, potassium carbonate or sodium carbonate or potassium hydroxide or sodium hydroxide to vinegar), nisin, potassium acetate, potassium, benzoate, potassium bisulphite, potassium diacetate, potassium lactate, potassium, metabisulphite, potassium sorbate, propionic acid, propyl -/ -hydroxybenzoate (propylparaben), sodium acetate, sodium benzoate, sodium bisulphite, sodium diacetate, sodium lactate, sodium metabisulphite, sodium propionate, sodium salt of methyl-p-hydroxybenzoate acid, sodium salt of propyl -/ -hydroxybenzoate. sodium sorbate, sodium sulphite, sodium dithionite, sorbic acid, sulphurous acid, calcium propionate, calcium sorbate, dimethyl decarbonate, natamycin, potassium sorbate, propionic acid, sodium diacetate, sodium propionate, sodium sorbate, sorbic acid, asorbic acid, ascorbyl palmitate, ascorbyl stearate, butylated hydro-xyanisole (a mixture of 2-tertiarybutyl-4- hydroxyanisole and 3-tertiarybutyl-4- hydroxyanisole), butylated hydroxytoluene (3,5- ditertiarybutyl-4-hydroxytoluene), citric acid, citric acid esters of mono- and diglycerides, L- cysteine, L-cysteine hydrochloride, gum, guaiacum, lecithin, lethicin citrate, monoglyceride citrate, monoisopropyl citrate, propyl, gallate, sodium metabisulphite, tartaric acid, tertiary butyl hydroquinone, tocopherols (alpha-tocopherol; tocopherols concentrate, mixed), nitrites (nitrates and nitrosamines), and combinations thereof. In some embodiments the meat analogue can include about 0.05% to about 10% preservative. In some embodiments, the meat analogue can include optionally about 0.1% to about 2% preservative. [0432] In some embodiments described herein, the food product comprises an antioxidant. The antioxidant can be selected from Dadex ARHS, ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), flavonoids, tannins, phenols, lignans, ascorbic acid, glutathione, melatonin, tocopherols, tocotrienols, uric acid, beta- carotene, lycopene, lutein, lipoic acid, vitamin E, selenium, manganese, and combinations thereof. In some embodiments, the meat analogue can include about 0.0001% to about 10% antioxidants. In some embodiments, the meat analogue can include optionally about 0.0001% to about 2% antioxidants.

[0433] In some embodiments described herein, the food product comprises a yeast. The yeast can be, active dry (traditional) yeast, instant yeast, bread machine/pizza yeast, rapid rise (or quick rising) instant yeast, fresh yeast, live yeast/probiotics, whole cell inactive yeasts, yeast fractions, yeast extracts, yeast cell walls, nutritional yeast, and combinations thereof.

[0434] In some embodiments described herein, the food product comprises a high moisture meat analogue.

[0435] In some embodiments described herein, the food product comprises a low moisture meat analogue.

[0436] In some embodiments described herein, the food product comprises a texturized protein.

[0437] In some embodiments described herein, the food product is selected from a flaked tuna analog, a chicken analog, a pulled pork analog, a fish analog, a sausage analog, a beef analog, a ground beef analog, and combinations thereof.

Method of Making a Meat Analogue

[0438] In some embodiments of the meat analogue composition herein, methods of making and/or producing the plant-based protein product of the present disclosure comprise processes including, but not limited to preconditioning (e.g., to hydrate a dry mix), mixing, conveying, extruding, compressing, cooking, heating, shearing, cooling, tenderizing, marinating, and/or drying.

[0439] The plant-based protein product of the present disclosure may be made and/or produced by introducing one or more protein sources, a plurality of dry ingredients, and a medium, along with any optional enhancer components (e.g., a flavoring agent) to an apparatus, an instrument, or an equipment, such as an extruding apparatus

[0440] In some embodiments, methods of making a texturized protein are provided.

A protein, a fat, an additive are combined into a mixture with water and extruded under heat and pressure. In an embodiment, the protein can be a Euglena- derived material. In an embodiment, the Euglena- derived material can be, for example, a Euglena beta glucan isolate. In an embodiment, the protein can be in an amount of about 0.01% to about 100% (e.g., about 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). In an embodiment, the fat can be, for example, a coconut oil. In an embodiment, the additive can be a flavourant. The flavourant can be in an amount of about of about 0.0001% to about 100% (e.g., about 0.0001, 0.001, 0.01, 0.1, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 98, 99, 99.5, 99.9, or 100%). The mixture can be heated to a temperature of about 80 degrees Fahrenheit to about 190 degrees Fahrenheit for 5, 10, 15, 20, 30, 60, 120 minutes or for about 1 hour to about 24 hours or longer.

[0441] Embodiments described herein are directed to a method of making a texturized protein comprising Euglena flour, the method comprising: combining one or more ingredients selected from a protein, a fiber, a fat, an additive, and other ingredients with water and Euglena flour to create a mixture; extruding the mixture under heat and pressure; and cutting the mixture with a die faced cutter, wherein a cutting speed (rpm) is variable.

[0442] In some embodiments, the cutting speed is about 500 rpm to about 1000 rpm. In some embodiment, the cutting speed is about 100 rpm to about 2000 rpm.

[0443] In some embodiments, the texturized protein is a large piece or plurality of large pieces about 1 cm or more in length (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or more cm in length).

[0444] In some embodiments, the texturized protein is a small piece or a plurality of small pieces about 1 cm or less in length (e.g., about 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 or less cm in length).

[0445] In some embodiments, the texturized protein is a high moisture meat analogue.

[0446] In some embodiments, the texturized protein is a low moisture meat analogue.

[0447] The compositions and methods are more particularly described below and the Examples set forth herein are intended as illustrative only, as numerous modifications and variations therein will be apparent to those skilled in the art. The terms used in the specification generally have their ordinary meanings in the art, within the context of the compositions and methods described herein, and in the specific context where each term is used. Some terms have been more specifically defined herein to provide additional guidance to the practitioner regarding the description of the compositions and methods. [04481 As used herein, the term “and/or” includes any and all combinations of one or more of the associated listed items. As used in the description herein and throughout the claims that follow, the meaning of “a”, “an”, and “the” includes plural reference as well as the singular reference unless the context clearly dictates otherwise.

|0449] All patents, patent applications, and other scientific or technical writings referred to anywhere herein are incorporated by reference herein in their entirety. The embodiments illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations that are specifically or not specifically disclosed herein. Thus, for example, in each instance herein any of the terms "comprising," "consisting essentially of," and "consisting of' can be replaced with either of the other two terms, while retaining their ordinary meanings. The terms and expressions which have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claims. Thus, it should be understood that although the present methods and compositions have been specifically disclosed by embodiments and optional features, modifications and variations of the concepts herein disclosed can be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of the compositions and methods as defined by the description and the appended claims.

[0450] Any single term, single element, single phrase, group of terms, group of phrases, or group of elements described herein can each be specifically excluded from the claims.

[0451] Whenever a range is given in the specification, for example, a temperature range, a time range, a composition, or concentration range, all intermediate ranges and subranges, as well as all individual values included in the ranges given are intended to be included in the disclosure. It will be understood that any subranges or individual values in a range or subrange that are included in the description herein can be excluded from the aspects herein. It will be understood that any elements or steps that are included in the description herein can be excluded from the claimed compositions or methods

[0452] In addition, where features or aspects of the compositions and methods are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the compositions and methods are also thereby described in terms of any individual member or subgroup of members of the Markush group or other group. [04531 The following are provided for exemplification purposes only and are not intended to limit the scope of the embodiments described in broad terms above.

EXAMPLES:

Example 1 - Euglena- Derived Dairy Vegan Yogurt Analogue Processes and Formulations

[0454] Yogurts can be made using for example, Euglena-derived materials, milk alternatives, sugar, acid, and water. A yogurt can have a final pH of about 3.0, 3.1, 3.2, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4., 4.5 4.6, 4.7 or more. Euglena- derived materials can make up about 5, 6, 7, 8, 9, 10, 12, 15, 18, 20, 25 % W/W or more of the ingredients or final product. Milk alternatives can make up about 60, 70, 80, 90, 95% W/W or more of the ingredients or final product. Sugar can make up about 4, 5, 6, 7, 8, 9, 10, 12, 15% W/W or more of the ingredients or final product. Acid, such as lactic acid can make up about 0.2, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. Oats or oat flour can also be added along with a milk alternative like oat milk at about 1.0, 5, 6, 7, 8, 9, 10, 15, 20% or more W/W. A vegan yogurt prototype, VI (Table 1) was created utilizing Euglena- derived beta-glucan isolate (“BGI”), oat milk, sugar, lactic acid powder, and water. Oat milk was chosen due to its low allergenicity. Starch was added to help thicken and stabilize the yogurt base system in combination with Euglena- derived BGI.

[0455] Inclusion of BGI was targeted to be 5 g per 170 g serving of yogurt or roughly 3% of the total formula. The target product formulation for the vegan yogurt incorporated 15-20% solids from BGI and 80-85% water. Oats and water were blended on high speed in a Ninja blender for 2 minutes. The mixture was strained using cheese cloth followed by addition of the other ingredients. The formulation was cooked for 20 minutes at 150 °F, and allowed to cool followed by refrigeration.

[0456] Lactic acid was added to reduce the pH to 4.4. Experiments were performed to test the impact of adding lactic acid before or after the heating step, to determine the amount needed to achieve the target pH of 4.4, and to determine impact on flavor. When lactic acid was added after the cooking step, the formulation did not thicken enough to become a yogurt. Even when the pH was dropped to 3.5 in increments, the formulation did not thicken but the flavor achieved the target sourness desired.

[0457] When lactic acid was added to the mix to pH 4.4 prior to cooking, the formulation thickened greatly and formed a gel similar to yogurt consistency. When the cooked formulation was allowed to sit overnight (10 hours) under refrigeration, it thickened too much and formed a pudding-like or Jell-O-like consistency. Water was added back to the refrigerated formulation to account for the 15% moisture lost during the cooking step. Vigorous stirring with a spoon was used to break up the gel network. The pH of the formulation produced by this process was 4.4 as targeted, but the flavor was judged to be too “oaty” and not sour enough to taste like yogurt. To improve the flavor, more lactic acid was added in increments and the flavor was tested for sourness. At pH 3.68, the desired texture and flavor of the formulation were similar to cultured dairy yogurt.

[0458] Moisture and shear, alone or in combination, can be used to help control the texture after cooking. Moisture can fluctuate during the process or can be added afterwards to achieve a desired texture. For example, the formulation could be thinned out with water to create a sauce or salad dressing consistency. Shear can also be applied to achieve the desired texture. The formulation still held its form even once after shearing was applied.

[0459] The amount of water to use to achieve the desired texture can be about 0, 1, 2, 5, 6, 7, 8, 9, 10, 12, 15, 20, 25, 30% W/W or more of the ingredients or final product of The acid, e.g., lactic acid can be added either before or after cooking.

[0460| At room temperature the BGI with water does not gel to the consistency of yogurt or thicker. When combined with lactic acid and heated to 180 °F it forms a thick gel. The incorporation of oats, which also contribute starch and a different kind of beta-glucan compared to the BGI, may also contribute to the gelling matrix.

[0461] For Euglena- derived vegan yogurt formulation V2, oats and water were blended on high speed in a Ninja blender for 2 minutes. The mixture was strained using cheese cloth and set aside. BGI and BGI water were added to a saucepan and stirred to combine. Sugar and oat milk were added and stirred. Lactic acid was added until pH 4.4 was achieved (0.13%). Everything was whisked together. The saucepan was heated on medium until the temperature reached 180 °F, while whisking to prevent burning, and held at that temperature for 10 minutes. After cooking the saucepan was placed in a cold water bath until the temperature reached 100 °F, then transferred to ajar with a lid and placed in the refrigerator to cool completely. The texture of formulation V2 was very thick after overnight refrigeration, setting to a Jell-0®-like (gelatin) or pudding-like texture. Water was added back to reduce the thickness of the formulation. The flavor was not sour enough, so lactic acid was added to reduce the pH to 3.68 (Table 2).

Table 1. Euglena derived Vegan Yogurt Analogue Formulation V 1 and V2

Table 3. Euglena- Derived Vegan Yogurt Analogue Formulation V3

[0462] Table 3 shows a potential formulation for Euglena- derived yogurt in which oats and water can be blended on high speed in a Ninja blender for 2 minutes. The mixture is then strained using cheese cloth and set aside. BGI and BGI water can be added to a saucepan and stirred to combine. Sugar and oat milk can be added and stirred. Lactic acid can be added until pH 3.7 is achieved. Everything can be whisked together. The saucepan can be heated on medium until the temperature reached 180 °F, while whisking to prevent burning, and held at that temperature for 10 minutes. After cooking the saucepan can be placed in a cold water bath until the temperature reached 100 °F, then transferred to a jar with a lid and placed in the refrigerator to cool completely.

[0463] In another formulation, Bob’s Red Mill oat flour and water was used in place of the oat milk with 15 % water addition for a total formula of 115 % to account for the moisture loss. The starting pH was 3.7 before cook. The final pH was 3.9 after cooking, after refrigeration and sitting approximately 20 hours. The texture of this formulation was thick like pudding. Shear and potentially water will be needed to break up the gel network. More lactic acid will be needed to drop the pH to 3.7 for flavor impact.

Table 4. Euglena- Derived Vegan Yogurt Analogue Formulation V4

|0464] Table 4 shows a formulation V4 for Euglena- derived yogurt, in which oats and water were blended on high speed in a Ninja blender for 2 minutes. The mixture was strained using cheese cloth and set aside. BGI and BGI water were added to a saucepan and stirred to combine. Sugar and oat milk were added and stirred. Lactic acid was added until pH 3.7 was achieved. Everything was whisked together. The saucepan was heated on medium until the temperature reached 180 °F, while whisking to prevent burning, and held at that temperature for 10 minutes. After cooking the saucepan was placed in a cold water bath until the temperature reached 100 °F, then transferred to a jar with a lid and placed in the refrigerator to cool completely. An example of the yogurt formulations described in Example 1 is found in FIG. 1. [0465] Additional non-dairy, non-allergen milk sources that can be used include sunflower or flax. Other milk sources include soy or almond milk in can be used place of oat milk. Water amounts and timing of addition, and lactic acid amounts and timing of addition can be varied. Addition of fruit such as strawberry or flavors can be added to impact flavor, texture, and pH. Additional liquid flavors such as vanilla can also be tested.

Table 5. Dry Basis

[0466] Table 5 shows the dry basis for ingredients.

[0467] Other formulations can be used to make yogurt analogs. Yogurts can be made using for example, Euglena-derived materials such as BGI powder, oils, lecithin, hydrocolloids, emulsifiers, water, and protein sources. A yogurt can have a final pH of about 3.0, 3.1, 3.2, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4., 4.5 4.6, 4.7 or more. Euglena- derived materials (e.g., BGI powder) can make up about 5, 6, 7, 8, 9, 10, 12, 15, 18, 20, 25 % W/W or more of the ingredients or final product. Oils (such as sunflower and coconut oils) can make up about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20% W/W or more of the ingredients or final product. Water can make up about 10, 20, 30, 40, 50, 60, 70, 80, 90% W/W or more of the ingredients or final product. Acid, such as lactic acid can make up about 0.2, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. Lecithin, such as sunflower lecithin can make up about 0.2, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. A protein, such as pea protein isolate can make up about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 20% W/W or more of the ingredients or final product. A hydrocolloid, such as carrageenan can make up about 0.2, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. An emulsifier can make up about 0.2, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. Two formulations were developed to prepare yogurt analogue using wet/dry BGI and wet or dry ready to gel beta glucan isolate. (See Table 6). This yogurt analogue had low fat and 8% protein.

Table 6. BGI-Based Yogurt Formulation [04681 BGI powder or slurry based yogurt. In this formation, a BGI slurry with 3% BGI (either by adding powder to water or diluting BGI slurry obtained from co-purification) was prepared, protein, sunflower lecithin, carrageenan into 50 g BGI slurry were added. The formulation was left to mix for 2 hours. An oil blend was prepared by mixing coconut and sunflower oil and add Myverol 18-92K emulsifier into it. The blend was heated to ensure the emulsifier was completely melted and mixed well with oil. The oil blend was poured into the BGI-protein blend and the mixture was homogenized using a polytron (omni GLH polytron mixer) at speed 9 for 5 minutes to form the emulsion. The pH of the emulsion was adjusted to pH 4 to 5 using lactic acid (FIG. 2A).

[0469] Wet RTG based yogurt. In this formulation a wet RTG slurry equivalent of 3% BGI in 50 g water was made, followed by the addition of protein, sunflower lecithin, carrageenan into the 50 g BGI slurry and left to mix for 2 hours. An oil blend was prepared by mixing coconut and sunflower oil and add Myverol 18-92K emulsifier into it. The blend was heated to ensure the emulsifier was completely melted and mixed well with oil. The oil blend was poured into the BGI-protein blend and the mixture was homogenized using a polytron (omni GLH polytron mixer) at speed 9 for 5 minutes to form the emulsion. The pH of the emulsion was adjusted to pH 4 to 5 using lactic acid (FIG. 2B).

Example 2 - Euglena- Derived Dairy Vegan Butter Analogue Processes and Formulations

[0470] Vegan butter can be made using for example, Euglena- derived materials, oils and/or fats, yeast extract, and salt and/or other flavors. Euglena- derived materials, such as Euglena flour and Euglena BGI can make up about 1, 3, 5, 7, 10, 15, 20, 30, 40, 50% or more W/W of the final product. Oils, fats, or combinations thereof can make up about 60, 70, 80, 90, 95, 99% W/W or more of the ingredients or final product. Yeast extract can make up about 0.1, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0% W W or more of the ingredients or final product. Salt or other flavors can make up about 0.01, 0.1, 0.25, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. The vegan butter formulation described herein had the consistency necessary to replace a natural butter. The vegan butter described herein had a properties similar to selected varieties of butter made from natural dairy materials such as the amount of protein, fat, sugar, sensory properties and/or a combination thereof.

[0471] In this study Vegan Butter was created using ingredients comprised of shortening, Euglena flour, and baker’s yeast extract. [0472] Initially a prototype utilizing vegetable/palm shortening, baker’s yeast extract and Euglena-dcmcd flour was created (Table 7). In this formulation the oil was melted and then the Euglena- derived flour and yeast extract were added. After stirring to combine, the formulation was refrigerated until starting to solidify. Before complete solidification of the fat, the blend was sheared to fully incorporate the sediment using a spoon or fork. The formulation functioned properly as a spreadable like butter analogue at room temperature but needed improvement for flavor. Euglena- derived beta-glucan isolate (“BGI”) was added for potential additional health benefits.

Table 7. Euglena- Derived Butter Analogue Formulation Prototype

[0473] Methods and Materials:

[0474] First, the oil was added to a pan; once melted, the Euglena flour and yeast extract were added and the contents were stirred to combine. The mixture was refrigerated until it started to solidify. Before complete solidification of the fat, the blend was sheared to fully incorporate the sediment using a spoon or fork.

[0475] It functioned properly as it was spreadable like a butter at room temperature but needed improvements for flavor. Euglena beta glucan isolate (BGI) was added for potential added health benefit claims.

[0476] Building on the prototype, _the VI Vegan Butter formula was developed incorporating an unmasked Euglena flour, adding in Beta Glucan Isolate (BGI), and using refined coconut oil with a melt temp of 80 °F in place of vegetable or palm shortening for sustainability and clean label reasons. Salt was also added for flavor improvement, as detailed in Table 8.

Table 8. Detailing the VI Formulation of the Vegan Butter

[0477] In the first trial of the V2 Formulation (V2a) of the Vegan Butter, a 10% masked flour was used in place of the original Euglena flour to see how flavor could be improved further. However, the masker used in this flour was too sweet for the application and a new formulation was tested (V2b).

[0478] Instead, V2b formula was developed using a masked Euglena flour with antioxidant incorporation, was evaluated along with garlic powder addition for flavor improvement. (Table 9, FIG. 3).

Table 9. Details the V2b Formulation of the Euglena- Based Vegan Butter

[0479] It is possible to add additional various flavours, herbs and spices to the Vegan Butter to develop differing flavour profiles and applications of the Euglena-<S&nv _Q & products.

Example 3 - Euglena Dairy Creamer Analogue Processes and Formulations

[0480] Creamer can be made using for example, Euglena- derived materials, oils, water, lecithin, hydrocolloids, emulsifiers, and sugar. Euglena- derived materials, such as Euglena BGI powder and Euglena BGI slurry can make up about 0.5, 1, 2, 3, 3, 5, 7, 10, 15, 20, 30, 40, 50% W/W or more of the ingredients or final product for the powder and about 50, 60, 70, 80, 90, 95, 98% W/W or more for the BGI slurry. Oils can make up about 1, 2, 3, 4, 5, 10, 20, 30 % W/W or more of the ingredients or final product. Lecithin can make up about 0.01, 0.1, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0 % W/W or more of the ingredients or final product. Hydrocolloids can make up about 0.01, 0.1, 0.25, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. Emulsifiers can make up about 0.01, 0.1, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0 % W/W or more of the ingredients or final product. Sugar can make up about 0.01, 0.1, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0 % W/W or more of the ingredients or final product. Where BGI powder is used water can make up 30, 40, 50, 60, 70, 80, 90, 95, 98 % W/W or more of the ingredients or final product.

[0481] Creamer formulations with high stability in hot coffee and minimal BGI sedimentation were developed (Tables 10 and 11). The formulation can be prepared using both wet and dry BGI. When the creamer was added to coffee, there was no cream layer formation on the top of the coffee for 30 minutes after the addition of creamer into the coffee and only minimal sedimentation of the BGI was observed after the addition of creamer into coffee (FIG. 4A and FIG. 4B). The creamer was highly pourable and there was not much change in viscosity during the refrigeration. Growth of fungus was observed in creamers after two weeks of storage in the refrigerator, however the creamer did not undergo any pasteurization before storage.

[0482| Method of Creamer Preparation Using BGI Powder. In this formulation, BGI powder was mixed with guar gum, sunflower lecithin in water and left to stir stirring for a minimum of 2 hours. The oil was mixed with Myverol 18-92K emulsifier and heated until the emulsifier was completely dissolved. Then the oil-emulsifier blend was added to the BGI blend and homogenized using a polytron blender (omni GFH polytron mixer) at speed 9 for 5 minutes to form the emulsion (creamer).

[0483] Method of Creamer Preparation Using BGI Slurry. In this formulation, the BGI slurry was diluted to obtain 3% BGI. Guar gum and sunflower lecithin were mixed into the BGI slurry and left to stir for a minimum of 2 hours. The oil was mixed with Myverol 18- 92K emulsifier and heated until the emulsifier was completely dissolved. Then the oil- emulsifier blend was added to the BGI blend and homogenized using a polytron blender (omni GFH polytron mixer) at speed 9 for 5 minutes to form the emulsion (creamer).

Table 10. Spray Dried BGI Based Creamer Formulation

Table 11. Wet BGI Based Creamer Formula

Example 4 - Euglena-Derived Cream Cheese Analogue Processes and Formulations

[0484] Cream cheese can be made using for example, various combinations of Euglena- derived materials, oils, water, lecithin, lactic acid, protein isolate, hydrocolloids, emulsifiers, lemon concentrate, pectin, and salt and/or other flavors, and optionally other ingredients. Euglena-AcmcA materials, such as Euglena ready to gel beta-glucan isolate Euglena BGI can make up about 1, 3, 5, 7, 10, 15, 20, 30, 40, 50% or more W/W of the ingredients or final product. Oils, fats, or combinations thereof can make up about 5, 10, 20, 30, 40, 50 % W/W or more of the ingredients or final product. Lecithin can make up about 0.1, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0% W/W or more of the ingredients or final product. Salt or other flavors can make up about 0.01, 0.1, 0.25, 0.5, 0.75, 1.0, 1.25, 1.5, 2.0, 3.0, 4.0, 5.0% W/W or more of the ingredients or final product. Protein isolate can make up about 1, 3, 5, 7, 10, 15, 20, 30, 40, 50% or more W/W of the final product. Lactic acid can make up about 0.1, 0.5, 0.75, 1.0, 2, 3, 4, 5 % or more W/W of the final product. Hydrocolloids can make up about 0.01, 0.1, 1.0, 2.0, 3.0, 4, 5, 6, 7, 8, 9, 10% or more W/W of the final product. Lemon concentrate can make up about 1, 3, 5, 7, 10, 15, 20, 30, 40, 50% or more W/W of the final product. Pectin can make up about 1, 2, 3, 4, 5, 7, 10, 15 % or more W/W of the final product. Two formulations were developed to prepare cream cheese analogues using BGI (Table 12) and RTG (Table 14). These cream cheese analogues were able to provide more than 2 g protein per 30 g serving.

[0485] BGI based cream cheese: Prepare BGI slurry with required amounts of BGI, pea protein, carrageenan, and sunflower lecithin. Leave the mixture for proper hydration of protein. Melt coconut oil and blend the emulsifier with it. Add the oil blend to BGI-protein blend and homogenize well. Refrigerate the mixture overnight and blend it again with lactic acid to reduce the pH of the mixture to 4.5. Then add flavours (parsley, garlic, lemon etc along with flavours given in Table 13) (FIG. 5A).

Table 12. BGI based cream cheese analogue formulation

Table 13. Flavor Additives

[0486] RTG based cream cheese analogue: In this method pea protein isolate, xanthan gum and gellan gums were dissolved in water first. The pH of the solution was around pH 7 (pH of pea protein isolate was pH 7). Then lemon concentrate was added to the protein- gum solution until the pH of the solution was around pH 3.5 to 4.5. The mixture was then gently stirred, large coacervates of pea protein and hydrocolloids were formed and started separating from the water phase. At this point, all the other ingredients were added to the mixture and mixed well. Sriracha and lime were added to improve the flavour of the cream cheese (FIG. 4B).

Table 14. RTG Based Cream Cheese Formulation

Example 5 - Euglena-Derived Cheese Sauce Analogue Processes and Formulations

[0487] Cheese sauce can be made using for example, various combinations of Euglena- derived materials, milk alternatives, yeast, proteins, oils, starch, hydrocolloids, and salt and/or other flavors, and optionally other ingredients. Euglena- derived materials can make up about 1, 3, 5, 7, 10, 15, 20, 30, 40, 50% or more W/W of the ingredients or final product. Oils, fats, or combinations thereof can make up about 1, 2, 3, 4, 5, 10, 20, 30 % W/W or more of the ingredients or final product. Protein isolate can make up about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10% W/W or more of the ingredients or final product. Salt or other flavors can make up about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30% W/W or more of the ingredients or final product. Starch can make up about 0.1, 0.5, 0.75, 1.0, 2, 3, 4, 5 % or more W/W of the final product. Hydrocolloids can make up about 0.01, 0.1, 1.0, 2.0, 3.0, 4, 5, 6, 7, 8, 9, 10% or more W/W of the final product. A Euglena- derived cheese sauce formulation is shown in Table 15. To make this formulation, all ingredients were sifted into the bowl of a food processor or high speed blender. They were mixed for 10 to 15 seconds to ensure all ingredients were blended. The resulting mixture was packaged in desired containers and serving amounts. To make into a mac and cheese dish, 6 cups of water and 1 tsp of salt were brought to a boil. Pasta was added and cooked, boiling for 6-7 minutes (can be adjusted for desired texture), stirring often. lOOmL pasta water was reserved. Pasta was drained, rinsed, and set aside. Reserved pasta water and seasoning packet were combined in a medium pot with 1 tbsp oil (Table 15). The water and seasoning were whisked until well combined and slightly thickened. The pasta was stirred in and cooked for 1-2 minutes, until heated through (FIG. 6).

Table 15. Euglena- Derived Cheese Sauce Analogue Formulation

Example 6 - Euglena-Derived Vegan Vanilla Yogurt Analogue Formulation

[0488] In the market there are multiple vegan yogurt products targeted at consumers. In these applications, the main ingredients consist of a milk alternative (nut, coconut, oat milk or powder), water, a sugar source, a starch source, a fat source, cultures, flavouring, protein, and/or a combination thereof.

[0489] In this study, the vegan yogurt formulation consists of oat flour, water, cocoa butter powder, sunflower lecithin, Beta Glucan Isolate (BGI) from Euglena, cane sugar, lactic acid, culture, and vanilla flavouring.

[0490] Methods and Materials:

[0491] The formulation that was used in this study for a Vanilla Vegan Yogurt is written in Table 16, which included the Euglena BGI at approximately 4%. The cocoa butter was added to a sauce pan and melted over heat. The sunflower lecithin was then added and the contents of the pan were stirred to combine. Once combined, the water was added to the pan and an immersion blender was used for one minute to combine: the contents foamed and turned white. The BGI and cane sugar were then added to the pan, again using the immersion blender to combine. Once incorporated, oat flour was then added to the contents of the pan, and the immersion blender was used to mix the ingredients together. Once incorporated, the contents of the pan were heated to 180 °F while whisking slowly. Once the temperature was reached, the stove top was turned off and the contents continued to be stirred for an additional two minutes. The saucepan was then transferred to a cold-water bath and left to cool to 100 °F, while continuing to whisk. The lactic acid and vanilla flavouring were then added, and the immersion blender as used to combine. A frozen culture was added and the contents stirred for five minutes; an immersion blender was be used. The finished product was then placed in a refrigerator to set for 24 hours to reach a firm finished texture (FIG. 7).

Table 16. Euglena- Derived Vegan Yogurt Analogue Formulation - Vanilla

Example 7 - Euglena-Derived Mango Vegan Yogurt Analogue Formulation

[0492] In the market there are multiple vegan yogurt products targeted at consumers. In these applications, the main ingredients consist of a milk alternative (nut, coconut, oat milk or powder), water, a sugar source, a starch source, a fat source, cultures, flavouring, protein, and/or a combination thereof.

[0493] In this study, the vegan yogurt formulation comprises of oat flour, water, cocoa butter powder, sunflower lecithin, Beta Glucan Isolate (BGI), cane sugar, lactic acid, culture, and mango flavouring.

[0494] Methods and Materials:

[0495] The formulation that was used in this study for a Mango Vegan Yogurt is written in Table 17, which included the Euglena BGI at approximately 4%. The cocoa butter was added to a sauce pan and melted over heat. The sunflower lecithin was then added and the contents of the pan were stirred to combine. Once combined, the water was added to the pan and an immersion blender as used for one minute to combine: the contents foamed and turned white. The BGI and cane sugar were then added to the pan, again using the immersion blender to combine. Once incorporated, oat flour was then added to the contents of the pan, and the immersion blender was used to mix the ingredients together. Once incorporated, the contents of the pan were heated to 180 °F while whisking slowly. Once the temperature was reached, the stove top was turned off and the contents continued to be stirred for an additional two minutes. The saucepan was then transferred to a cold water bath and left to cool to 100°F, while continuing whisk. The lactic acid and vanilla flavouring were then added, and the immersion blender was used to combine. A frozen culture was added and the contents stirred for five minutes; an immersion blender was used. The finished product was then placed in a refrigerator to set for 24 hours to reach a firm finished texture.

Table 17. Euglena- Derived Vegan Yogurt Analogue Formulation - Mango

[0496] Ingredients: Water, Oat Flour, Cane Sugar, BGI, Cocoa Butter, Natural Flavor, Lactic Acid, Sunflower Lecithin, Cultures

[0497] Other formulations of the vegan yogurt were tested and formulation of a vegan yogurt could use any of the following: using quick oats, bran oats, BGI, BGI Slurry, cane sugar, lactic acid, and/or flavouring in various ratios and combinations thereof. BGI can be added as a dried ingredient or as a wet ingredient. The BGI can be added directly from downstream processing as a wet ingredient. Wet biomass maybe added to from the yogurt. Euglena biomass, Euglena flour (protein and/or beta-glucan rich), Euglena protein concentrate, Euglena protein isolate, and Euglena beta-glucan isolate can be used individually or in combination, as a dry or wet ingredient in combination with each other.

[0498] Vegan Yogurt Formulation Summary: The vegan yogurt formulation described herein had the consistency necessary to replace a natural yogurt. The vegan yogurt described herein had a property similar to a selected from a group consisting of the amount of protein, fat, sugar, dietary fiber (DF), sensory properties and/or a combination thereof. Example 8 - Euglena based vegan cheese

[0499] Cheeses can be made using, for example, Euglena- derived materials, A Euglena- derived material, an oil, an antibacterial agent, an antifungal agent, a hydrocolloid, water, and optionally a yeast. The Euglena- derived material can make up about 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 18, 20, 25 % W/W or more of the ingredients or final product. An oil can make up about 10, 15, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50% WAV or more ofthe ingredients or final product. An antibacterial agent can make up about 0.01%, 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50% WAV or more ofthe ingredients or final product. The antifungal agent can make up about 0.01%, 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50% W/W or more of the ingredients or final product. A hydrocolloid can make up about 1, 5, 10, 15, 20, 25, 30, 25% W/W or more of the ingredients or final product. Water can make up about 1, 5, 10, 15, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 60, 70, 80% W/W or more of the ingredients or final product. A yeast can make up about 0.5, 1, 1.5, 2, 2.5, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25% W/W or more of the ingredients or final product. The vegan cheese formulation described herein had the consistency necessary to replace a natural cheese. The vegan cheese described herein had a property similar to selected varieties from a group consisting of the amount of protein, fat, sugar, sensory properties and/or a combination thereof.

[0500] In this study, the VI formulation was tested using ingredients comprising of water, coconut oil, Ticagel CA 0979M, Euglena Protein Concentrate, salt, potassium sorbate, liquid cheddar flavour. The V3 formulation was tested using water, coconut oil refined, Ticagel®, Euglena protein concentrate, salt, yeast extract, Galacin Nisin NaCl 2.5, Galacin Natamycin 201 E235, and liquid cheddar flavour.

[0501] Methods and Materials for V 1 :

[0502] The formulation that was used in this study for a Vegan cheese is written in Table 18, which included the Euglena protein concentrate at approximately 5%. The dry ingredients were pre-blended prior to adding the water (ambient) to the Thermomix. The dry blend was then added and mixed at speed 3. Once combined, melted coconut oil and liquid cheddar flavour were added to the Thermomix. It was heated to 180 °F and held for 7 minutes at speed 2. It was hot filled into containers, and stored at 40 °F, and evaluated after 7 days.

[0503] Observations on day one the texture was gummy looking or lumpy, like the fat was not dispersed. [0504] Observations one week later showed a firm but cut-able (or sliceable) product, similar to a block of cheese. It was somewhat spreadable, but crumbly and dry similar to goat cheese (FIG. 8).

[0505] In this formulation, there was a 3.60% Euglena protein inclusion, with 1.08 g of protein in a 30 g serving size.

Table 18. Vegan Cheese Formula VI

[0506] Upon observation, VI functioned great for a hard cheese. It shreds and cuts nicely. A small piece was melted in the microwave to melting, about 20 seconds. The result was a somewhat gummy texture and fat seeped out which was in line with the market products as well. It didn’t taste like Euglena and the color was acceptable. It needed a little more flavor for a more complex profile. A yeast extract was added to help and this version was labeled as Vegan Cheese Formula, V3 (Table 19).

[0507] Methods and Materials for V3: The formulation that was used in this study for a Vegan cheese is written in Table 19, which included the Euglena protein concentrate at approximately 4%. Pre-blend all dry ingredients, prior to adding water to Thermomix. Then add the dry blend to the Thermomix and mix at speed three. Add melted coconut oil and liquid flavour to the Theromix. Heat mixture to 180 °F and mix for seven minutes at speed 2. Once complete, hot fill into containers, store at 40 °F and evaluate after seven days.

Table 19. Vegan Cheese Formula, V3

[0508] Preservatives were added to V3 and the samples were vacuum packed after one week. After one week the vegan cheese was observed to shred well, and have a softer feel, flavour was improved. When melted, the texture was gummy and fat seeps out.

[0509] Vegan Cheese V3 was the same as VI but with yeast extract addition for flavor. This was a well rounded flavor profile that masks the Euglena fully. This was the final formula for a hard vegan cheddar cheese going forward.

Example 9 - Softer Euglena based vegan cheese analogue

[0510] In this study, V2 of the Vegan Cheese was created by combining water, refined coconut oil, sunflower oil, Ticagel®, Pea Protein, Euglena protein concentrate, salt, yeast extract, liquid cheddar flavour, lactic acid powder and potassium sorbate.

[0511] Methods and Materials: The formulation that was used in this study for a Vegan cheese is written in Table 20, which included the Euglena protein concentrate at approximately 3%. Water (ambient) was added to the mixing bowl, and the speed was turned to 2 and Ticagel was added, and the proteins were dry blended while mixing. Melted fat was added (100 °F) gradually while mixing. The remaining ingredients were added: salt, seasoning, preservatives, flavour and colour while mixing. When homogenous, the mixture was slowly heated to 185 °F at speed 4 and held for 3 minutes at 185 °F. The mixture was hot filled into containers and stored at a refrigerated temperature for up to 7 days to obtain proper hardness, texture and mouth feel. [0512] Observations for day the cheese was made included a nacho cheese like appearance and a pungent, cheesy flavour.

[0513] Observations after one week showed a softer cheese texture, similar to Velveeta that was spreadable and gooey. The cheese flavour was good, however there were some off notes in the flavour (FIG. 9). In terms of protein, 2.84% protein from pea protein, 2.84% from Euglena protein concentrate for atotal of 5.68% for 1.704 grams of protein in 30 gram serving.

Table 20. Vegan Cheese Block Formulation 2, Softer Cheese

Example 10 - Additional Beta Glucan Isolate Based Vegan Cream Cheese Formulations [0514] Introduction: The global market of plant-based alternatives to dairy was estimated to be USD 22.6 billion in 2020. And it is projected to reach USD 40.6 billion by 2026. This fast growth is due to the increasing prevalence toward allergenicity of cow’s milk, lactose intolerance, changing consumer lifestyles, and interest in alternative diets (e.g. vegan and flexitarian). Moreover, the COVID-19 pandemic accelerated this process because it drove consumers to reconsider their lifestyle and choose a more plant-based diet as a healthier option. Thus, the demand for a variety of dairy alternatives to vegan yogurt, vegan cream cheese, vegan cheese, vegan butter, and others, is gradually gaining importance in the market nowadays. [05151 Titanium Dioxide (T1O2) is a common whitener in foods. It has only one food functionality, which is making a product look whiter. However, the European Food Safety Authority deemed that titanium dioxide is unsafe in foods in March 2020. Beta-Glucan Isolate (BGI) from Euglena has the similar functionality that is an all-natural healthy alternative to titanium dioxide applied in food products, especially dairy products or alternative dairy products.

[0516] Soybean was commonly used in vegan dairy products. However, one potential limitation of soybeans are the concerns with high estrogen levels and being a common food allergen. Moreover, soybeans in particular exhibit flavor and mouthfeel challenges, specifically, beany flavor and gritty mouthfeel. In this study, pea protein and other proteins were used instead of soybeans.

[0517J Objective: The objective of this study was to test how Euglena based BGI improves the whiteness of vegan cream cheese and to test how different protein sources affect sensory properties of vegan cream cheese.

[0518] Materials & Methods: Materials used in the formulations of a vegan cream cheese can include: Water, lactic acid, pea protein, fava protein, lentil protein, carrageenan, Euglena BGI, plant-based butter, sunflower lecithin, Myverol 18-92K, cream flavor.

[0519] Step 1 Making water mixture:

[0520] Lactic acid was dissolved in water. Protein(s) used are then dissolved in water and the lactic acid mixture and left for proper hydration for about 2 hours. Next, the carrageenan and Euglena BGI were added to the mixture while stirring.

[0521] Step 2 Making oil mixture:

[0522] For the oil mixture, sunflower lecithin, plant-based butter, Myverol 18-92K were heated to above the melting point (i.e. heated until fully melted) and mixed well.

[0523] Step 3 Combination:

[0524] The oil mixture and natural cream flavor were added to the water mixture and blended well using a blender until uniform.

[0525] Results:

Table 21. Cream cheese Formula 1

[0526] The vegan cream cheese that was generated from Table 21 was a little bit more yellow than regular cream cheese (FIG. 10). It also had a little bit of a gritty mouthfeel, however the hardness and spread ability were like regular cream cheese.

Table 22. Cream cheese formula 2

[0527] The vegan cream cheese that was generated from Table 22 was whiter than cream cheese formula 1 because more BGI (more than 10%) was used (FIG. 11). It looked like regular cream cheese, was creamy and a little bit gritty. Except for the slightly gritty mouthfeel, the texture was like regular cream cheese. The hardness and spread ability were just like regular cream cheese and had a creamy flavor.

Table 23. Cream cheese formula 3

[0528] The vegan cream cheese that was generated from Table 23 had an appearance that was like cream cheese made with pea protein (FIG. 12). However, the texture was softer than the cream cheese made with pea protein. It was also very smooth.

Table 24. Cream cheese formula 4

[0529] The vegan cream cheese that was generated from Table 24 had the surface of the cream cheese turned pink very easily (FIG. 13). That may be because this ingredient (lentil protein) contains some other substance that can be oxidized easily. It formed the right texture and was very smooth. It had a little bit of a beany flavor but it's still acceptable with a cream (dairy) flavor added. The bitterness may be due to the oxidation.

[0530] Conclusion

[0531] The Euglena BGI significantly improved the whiteness (colour) of the vegan cream cheese. The vegan cream cheese made with pea protein had the better hardness and spread ability compared to the vegan cream cheese made with fava protein and lentil protein. The hardness and spread ability of vegan cream cheese made with pea protein was similar to the regular cream cheese. However, it had a gritty mouthfeel.

Example 11 - Emulsification of BGI and different form

[0532] Introduction: Solid particles used to stabilize the o/w (oil in water) type of emulsion were discovered a century ago and have undergone extensive study in recent decades. While there are a few advantages to the solid particle emulsion stabilizer such as: reduction in the possibility of coalescences, giving greater stability to emulsions, having useful characteristics through the solid particle; and the use of food-grade solid particles in the emulsion. [0533] While many of the solid particles have been studied to prepare Pickering emulsion, such as silica, clay, nanoparticles, chitosan, cyclodextrin, and wax esters, among others, in this study we have chosen to focus on natural polysaccharide granules, paramylon granules, which were found to be suitable for Pickering emulsion.

|0534] Instruments: In this study, two types of homogenizers were used, a low power homogenizer (Fisherbrand 150 handheld homogenizer) and a higher power homogenizer (Omni international GLH-01), and their specializations are listed below in Table 25.

Table 25. The specializations ofhomogenizer

[0535] The majority of the study was conducted using the Higher power Homogenizer with a setting at 6 where the speed was 28,000RPM and homogenized for 5 min. However, it was confirmed that the BGI emulsion was also prepared when the Lower power homogenizer (Fisherbrand 150 handheld) with a speed setting at 35,000RPM and homogenized for 5 min.

[0536] When using the high power homogenizer, Beta Glucan Isolate (BGI) was incubated in oil for 24 hours, and then the emulsion was prepared. The oil incubation time (4 hours, 24 hours, 48 hours, 96 hours and 10 days) of BGI effect on emulsion formation was evaluated with the low power homogenizer. Our study indicated that a longer oil incubation time was required when a lower power homogenizer was used.

[0537] Materials: Spray dried BGI (SBGI), spray dried biomass, freeze dried milled paramylon (MP), freeze dried fully milled paramylon (FMP) and partially milled paramylon (PMP) are from Noblegen; palm oil, canola oil, linseed oil, sunflower oil and coconut oil are from the local market. The FMP was prepared for BGI with 10% solid content using a CMC (Consumer milling & Consulting Inc.) 1.5L horizontal media mill with two-gallon stainless steel mixing vessels charged with a 75% load of 0.4-0.6 mm Zirconium oxide grinding beads. The mill circulation range was 9.1 gal hr ¹ , and milling time was determined by viscosity. After approximately 2.5 h, the viscosity approached the maximum machine allowance and the milling was considered complete to give FMP. When the solid content was 15% for the milling process, a PMP was given. Please note that the milling portion in PMP was not determined.

[0538] Material Adding Sequence: BGI was added using different sequences to prepare the emulsion. Method (1): the BGI was incubated in water overnight (16 hours), and then oil was added, followed by homogenization using the high power homogenizer with speed of 28,000 rpm and homogenized for 5 mins. Method (2): the BGI was incubated in oil for overnight (16 hours), and then water was added, followed by homogenization using the high power homogenizer, as seen in FIG. 14.

[0539] Our study found that BGI emulsion was successfully prepared from method (2), but not from method (1). Although the oil-water emulsion was detected in method (1), the BGI was not incorporated into the emulsion, while BGI was spread in the water phase. Images of the emulsions are present in FIG. 15, below. FIG 15B was prepared using Method (1) and Fig 15(C) was prepared using Method (2).

[0540] Applied Mixing Forces: In this study, three mixing forces were used during the emulsion preparation: Vortexing, low power homogenizer, and a high power homogenizer. It was found that the mixing force needed for formation of BGI emulsion was related to the incubation time of BGI in oil. A longer incubation time lowered the mixing force required. For example, the BGI emulsion was formed with a vortex when the BGI as incubated in oil for 10 days. When using the low power homogenizer, the BGI needed to be incubated in oil for more than 48 hours; while with a high power homogenizer, the BGI was only incubated in oil for 16-24 hours, as seen in FIG. 16. The BGI emulsion was formed using low power homogenization when the BGI was incubated in oil for 24 hours, however it was not stable under a shearing force or dilution. To achieve a stable emulsion, a longer incubation time (such as 48hours+ for freeze dry samples of BGI, but 96hours+ for spray dry samples of BGI) was needed

[0541] Sample Effects: In this study, the samples used consisted of spray-dried beta glucan isolate (SBGI), freeze dried beta glucan isolate (FBGI), fully milled paramylon (FMP) that was milled with 10% solid content, partially milled paramylon (PMP) from that was milled with 15% solid content, and spray dried protein-rich biomass/flour. Emulsions were formed with all samples, however there was a difference observed in the micelles of the emulsion. For BGI, biomass, and PMP samples, the micelles contained the samples within micelle bubble, however for FMP it was not detected. As well, spray dried BGI required longer oil incubation time than freeze dried BGI to prepare a stable emulsion, as seen in FIG. 17. [0542] Oil Incubation Time: It was found that oil incubation, at room temperature, was essential to prepare a dense and stable BGI emulsion, especially using a low power homogenizer. With the high power homogenizer, a 24 hour incubation period time was sufficient for SBGI to prepare a dense and stable BGI emulsion, however the incubation time was 10 days for SBGI, and 4 days for FBGI to produce dense and stable emulsions when a low power homogenizer was used, as seen in FIG. 17. Interestingly, the emulsions prepared with a short incubation time were separated into two phases after dilution: top phase (TP, or lower density phase) and bottom phase (BP, or higher density phase), as seen in FIG. 18. With increasing BGI incubation time in oil, the emulsion was stable in one phase. This was likely due to the changes of the hydrophobic properties of the BGI with the incubation time increase, as seen in FIG. 19.

[0543] BGI Emulsion Formation Processes: Four types of BGI incorporated emulsions were found during our BGI emulsions preparation, as seen in FIG. 20.

[0544] Type 1 was found in the emulsion that was prepared with the shortest oil- incubation time (<16 hours), where the BGI was not efficiently bonded with oil, and a large portion of BGI was easily dispersed into water after homogenization. In type 1, BGI was included inside of the oil drop by the homogenization forces, and the size and distribution of oil drops are similar to those without BGI, as seen in FIG. 21. Compared with the emulsion, the emulsion micelles with BGI incorporated are larger (as seen in FIG. 20B, FIG. 20C, FIG. 20D). The emulsion micelles in this type were not stable, and oil droplets will be aggregated to larger droplets.

[0545] The Type 2 and Type 3 emulsions were found in emulsions that were prepared with an oil-incubation time of 24, and 48 hours, respectively. The emulsion micelles were separated easily into two parts when diluted: lower density portion (OIL+BGI phase) and higher density portion (Water +BGI phase). As seen in FIG. 20, in the lower density portion of type 2 (FIG. 20B), BGI seemed to function as an emulsifier surrounding the oil droplets (forming Pickering emulsions), but the density of BGI was lower, compared to Type 3 (FIG. 20C), in which portion of oil droplets contain higher density of BGI. In their higher density portion, few emulsion micelles were found in type 2 emulsion (FIG. 20F), but much more BGI emulsion micelles were found in type 3 (FIG. 20G). Type 4 emulsion was given when BGI was incubated in oil for a longer time (such as 96 hours). This type emulsions were stable and dense. All of the emulsion micelles of type 2 to 4 were stable for more than 2 weeks at room temperature (not determined for a long time). [05461 It should be noted that both type 2 and type 3 had better dispersion when they were poured into water, while type 4 was more like oil drops when poured into water. In this context, the emulsion of type 2 and type 3 could be better applied as coffee creamer.

[0547] Mechanism of BGI emulsion formation: According to the phenomena during formation of BGI emulsion, BGI emulsions were formed following the mechanism: BGI was bonded with oil through hydrogen bonding, making it more hydrophobic, and then BGI- oil cluster acted as oil phase when homogenization was applied to form O/W BGI emulsion micelles. When the BGI was fully bonded with oil, a highly dense and stable emulsion micelle was formed where BGI was not only present on the surface of micelles, but also inside of micelles. However, when the BGI was partially bonded with oil, the BGI granule had balanced hydrophobicity and hydrophilicity (HLB), making BGI granules suitable as an emulsifier for a O/W Pickering emulsion.

[0548] Effect factors of the BGI Emulsions

[0549] BGI content: In this study, the colour, thickness and phase stability of emulsions were visibly observed, and the emulsion micelles were imaged using a microscope. It was found that the BGI was included in emulsion micelles only when BGI was premixed with Oil.

[0550] With similar amounts of water and oil content, the whiteness in colour, thickness in texture, and stability in gravitational separation of emulsions increase, while the emulsion size decreases, with an increase in the amount of BGI.

[0551] The size of emulsion micelles becomes less even with the increase of BGI amount or BGI/Oil ratio as indicated in FIG. 22. Please note that the emulsion micelles can be stable for up to, and over, one month. The microscopic images suggest that the BGI granules effectively prevented the phenomena of flocculation and coalescence that lead to the brake- down of an emulsion.

[0552] The emulsions with lower BGI were also investigated. Maintaining the weight ratio of Oil and Water as 1:4, the BGI amount was varied from 0.25% to 2.5%. It was found BGI Pickering emulsion was formed when the including amount of BGI was up to 0.6%, as seen in FIG. 23.

[0553] As seen in FIG. 23, From 0.6%to 1.2% BGI content, the size of emulsion was not significantly changed. However, the size was significantly decreased from 1.2% to 2.5% BGI content.

[0554] Oil content: In this study, the effects of oil content was explored based on the BGI in Canola oil (FIG. 24). The BGI in Emulsion was maintained as 10%, while canola oil (Ca) varied from 10% to 70%. The samples were labeled as Ca-10 (10-10-80), Ca-20 (10- 20-70), Ca-30(10-30-60), Ca-40(10-40-50), Ca-45(10-45-45), Ca-60(10-60-30) and Ca-70(10- 70-20), respectively, according to oil content. The emulsion micelle was formed in Ca-10 to Ca-60 efficiently, but not in Ca-70 due to high viscosity. The size of the emulsion micelle and its phase stability (here mainly for gravitational separation) increased with the increase of oil content. The white emulsions appeared as a thin creamer in Ca-10, to a thicker creamer in Ca- 20, to a drinkable yogurt in Ca-30 and Ca-45, and to a Greek yogurt in Ca-60. It should be noted that the yogurt-like emulsion in Ca-60 was formed after a few seconds of homogenization, and the homogenizer was blocked by the emulsion due to its high viscosity.

[0555] Oil types: The effect of oil type on the emulsion was investigated by using five oils with a variation in melting point, composition, density and viscosity (Table 26). The oils chosen for this study were Canola oil (Ca), Palm oil (P), Sunflower oil (Su), Linseed oil (Li) and Coconut oil (Co), and their information was listed in Table 2. Two compositions of the emulsion were used in this study with a ratio of BGI: Oil: Water as 10: 10:80 and 10:45:45. Firstly, the amount of BGI and oil was added to a 50 mL centrifuge tube and fully mixed, they were kept overnight at room temperature, except for Palm oil and Coconut oil which were stored at 40°C due to their higher melting point.

Table 26. Oils used in this study and their compositions

10556] The emulsions were prepared using Omni international GLH-01 homogenizer at the speed of 28,000 rpm and homogenized for 5 mins, and the microscopic images were presented in FIG. 25.

[0557] The effect of oil types on the color, thickness, emulsion micelle size, and emulsion phase stability was investigated. The results were as follows:

[0558] Color: The color of the emulsion was determined by physical observation. It should be noted that the color of BGI in oil was white to off-white and in some cases, light yellow, before homogenization. However, all emulsions were white in color. The whiteness of the emulsions in the order of Co-10(45)>Su-10(45)~Ca-10(45)>P-10(45)>Li-10(45), and the color in emulsions of xx-10 was whiter than in xx-45.

[0559] Thickness: The thickness was determined by physical observation. In freshly prepared emulsions, the order of the thickness from thickest to thinnest was Li- 45~Su-45>Ca-45>P45>Co45. However, when left standing over time, the Co45 became thicker, and P45 became thinner. Upon standing, the order changed to Co45>Li-45~Su-45~Ca- 45>P45. It should be noted that it was difficult to visualize the differences in the xx- 10 samples as their textures were more creamer-like, while those of the xx-45 s were similar to that of a yogurt drink.

[0560] 3. Phase stability

[0561] The phase stability of the emulsion was physically observed and recorded in the different time periods. At 4 hours, SulO and Li 10 were not visible to have phase separation, CalO started phase separation, and Co 10 and P10 showed phase separation at around 10 to 20% in 4 hours. The order of the phase stability from most stable to least stable was Sul0~Lil0>Cal0>Col0>P10, however, after overnight (16 hours) standing, the order was Sul0>Lil0>P10~Col0>Cal0. For the samples of xx-45, the phase remained stable after 2 days.

[0562] Size of emulsion micelle: The emulsion micelle was imaged using microscopy with a 40X magnification, as seen in FIG. 25. It was found that the emulsion micelles in Canola oil, Palm oil, and sunflower oil had similar size, smaller than linseed oil, and Coconut oil emulsion had the largest micelles size. [05631 Stability of Emulsion micelle: The emulsion micelles were imaged at 2 hours, overnight (16 hours) and 4 days. There was no evidence showing that the emulsion micelles were not stable, even after 2 weeks.

[0564] Additives: Vinegar and Lime juice were applied to evaluate their effect on BGI emulsion formation. In this study, a 10% vinegar water solution, and a 100% Lime juice were used to replace water during emulsion preparation to supply acetic acid and citric acid, respectively. The BGI and Canola oil contents were maintained as 10%. The emulsions were prepared using method (2), where BGI was incubated in oil overnight, and then water was added, followed by homogenization using a high power homogenizer at speed of 28,000 rpm and homogenized for 5 min. The samples were labeled as V2 (VlO-10-80, 10% BGI-10% Canola oil-80 vinegar solution), and L2 (LlO-10-80, 10% BGI-10%Canola oil and 80% Lime juice). In comparison, the samples were also prepared using method (1), where BGI was soaked in water first and then mixed with oil, followed by homogenization. The samples prepared using method 2) from vinegar and lime juice were labelled as V3 and L3, respectively. And the emulsions without BGI were labeled as V 1 and L 1 , respectively. It was found that the emulsion micelles in V2 were similar to Ca-10 (C2, the control), but the micelles in L2 were lower density and smaller in size than Ca-10 (C2) and V2.

[0565] In sample V3 and L3, an Oil-water emulsion was formed, but BGI was not in the emulsion micelle. Instead, it was suspended in a water phase. However, the existence of oil-water emulsion enhanced the stability of the BGI suspension phase. No apparent phase separation was seen within 6 hours when Vinegar and Lime juice were added

[0566] Stability

[0567] pH Effect: In this study, the emulsions were prepared using the Lisherbrand 150 handheld homogenizer with the speed of 35,000 RPM and homogenized for 5 min. Prior to homogenization, BGI was incubated in canola oil for 10 days.

[0568] 0.5 mL of BGI emulsion (BGPOil: Water=10:20:70) was added into 1 mL water with pH 0.8, 2,5,7, and 8 to evaluate the emulsion stability in different pHs. Acidic solution was prepared from hydrochloric acid (HC1), and basic solution was prepared from 1M sodium hydroxide solution. BGI emulsion was at a different pH for 10 min, 60 min, 4 hours, and 24 hours, microscopic images taken during these time periods, and it was found that the emulsions are stable at pH range from 0.8-8 at the detected time.

[0569] Storage temperature: In this study, the emulsion was prepared from Canola oil (Ca), Coconut oil (Co), Palm oil (P), Linseed oil (Li) and sunflower oil (Su) using Omni international GLH-01 with speed of 28,000 rpm and homogenized for 5 mins. Prior to homogenization, BGI was incubated in oil overnight (16 hours) at room temperature (Canola oil, Linseed oil and Sunflower oil) or 40°C (palm oil and coconut oil due to their higher melting point). Six Canola oil emulsion compositions were used in this study. They are BGLoil: water as 10-10-80 (CalO), 10-20-70(Ca20), 10-30-60 (Ca30), 10-40-50(Ca40), 10-45-45 (Ca45) and 10-60-30 (Ca60). For other oils, 10-10-80 (xxlO) and 10-45 -45 (xx45) were used as emulsion composition.

[0570] The emulsions were stored at -20 °C, 4 °C, room temperature overnight, and at 80°C for 2 hours. It was found that the emulsion micelles were broken after being stored at -20 °C overnight, but no clear evidence showing that they were broken at other temperatures, including 80°C.As an example, the microscopic images of 10-30-60 Canola oil emulsion were presented in FIG. 26.

[0571] In Hot Water: 200 μL emulsions (CalO: BGFCanola oil: water in a 10:10:80 ratio and Ca45: BGFCanola oikwater in a 10:45:45 ratio) that were prepared above were added into 10 mL hot water (~70°C). It was found that the emulsion micelles remained stable after stirring in the hot water, as confirmed by microscopy. After standing for 2 hours at room temperature, the emulsion micelles accumulated at surface of the water. This indicates that if used as a coffee creamer for example, when added to hot coffee, the emulsion (what was acting as a creamer), remained stable and did not break.

[0572] Time: BGI emulsions were stable at room temperature for up to 2 weeks, as checked by microscopy. As they were stable, studies on longer time frames were not conducted.

[0573] Textures of the emulsions: In terms of texture, the emulsion varied from consistencies similar to milk, cream, yogurt and Cheese, as seen in FIG. 27. In call cases, the BGI was soaked in oil overnight for 16 hours. It was then homogenized using Omni international GLH-01 with speed of 28,000 rpm and homogenized for 5 mins. In order to get the different textures, it was the ratio of BGI to oil to water that determined the final texture consistency. As well, it should be noted that the mixtures of BGI/OIL/Water before homogenization can also be milk, Cream, Yogurt, or cheese. The thickness of the sample increased with homogenization. For example, milk-like texture before homogenization changed into 2-5% Cream after homogenization, and the 2% -5% cream into a texture similar to a 35 % cream, a 10% cream, or into drinkable yogurt; 35% cream into a Yogurt; and a drinkable yogurt into cheese. It should also be noted that the homogenization became less effective when the cheese-like texture was generated as the homogenizer became blocked. [0574] In terms of phase stability (gravitational separation), emulsion phase stability increased with the increase of its thickness. The milk-like emulsions had phase separation within 20 minutes, 1-3% cream -like emulsions had phase separation within 4 hours, but 5% cream-like emulsions or more thick emulsions were stabilized for more than 24 hours. The emulsion micelles were imaged for more than 2 weeks and it was found that the emulsion micelles were stable past the two week timeframe.

[0575] It should be noted that a stabilizing agent, for example, pectin can help phase stabilization. To confirm the function of pectin, emulsions with composition of 0-1% pectin- 1% BGI -10% oil were prepared using Omni international GLH-01 with speed of 28,000 rpm and homogenized for 2 minutes using method 2 above, but BGI was not incubated in oil. The results show that stability of emulsion phases increases with the pectin increases. Microscopic images confirmed that both pectin and BGI involved the emulsion formation, and the emulsion size decreases with increase of pectin amount.

[0576] Sensory tasting of BGI emulsions: Emulsions were prepared using both method 1) and 2) as described in the additive section. The BGI content was maintained as 10%, but Canola oil contents were 10% and 45%. Three samples were prepared and a sensory test was conducted. The samples, Ca-lOa (10-10-80) and Ca-45a (10-45-45) were prepared using method 2), and Ca-lOb (10-10-80) using method 1).

[0577] It was found Ca-lOa appeared similar to a 2-3.5% milk, but tasted closer to a 5-50% Creamer, Ca-45a was similar to a salad dressing or thicker creamer, resulting in the BGI emulsions having a cream-like mouth feeling. Compared to Ca-lOa and Ca45a, Ca-lOb appeared as 1% milk and tasted watery.

[0578] Discussions and Conclusions

[0579] Oil in water emulsions have been used in many commercial products, such as food, supplements, personal care, cosmetics, detergents and in the pharmaceutical industry. However, the emulsion is a thermodynamically unstable system, therefore a suitable emulsifier is necessary to stabilize the emulsion. Green, environmentally friendly, and non-toxic natural emulsifiers have been investigated widely with some natural emulsifiers such as small molecular surfactants or proteins, are easy to form small droplets, but are not stable at highly acidic conditions (pH<3) or high ionic strengths. Some, like polysaccharides, are stable under those conditions, although they require a high surfactant-to-oil ratio.

[0580] Micro Crystalline Cellulose (MCC) or Cellulose nanoparticles are a very efficient solid particle emulsifier to form emulsions containing 10-20% oil, and 2.5-5% MCC. [05811 In this study, emulsions were successfully prepared with paramylon granules (PGs) as emulsifiers. The emulsions contain about 5% to about 70% oil and 0.6% to about 40% PGs. PGs are not only used as emulsifiers but can also be encapsulated in emulsion micelles as nutrients. The emulsion varied from common Pickering emulsions to high internal- phase emulsions (HIPEs) by varying the contents of PGs and Oils, functioning as cream, Yogurt, and/or Cheese- analogues. HIPEs in the food industry look to be quite promising and offer an innovative approach for converting lipid oils into solid like fats without using saturated or trans fatty acids.

[0582] Additionally, BGI emulsion can be prepared using low energy mixing, such as vortex, and/or a low power (150W) mixing homogenizer. Given that emulsions normally need a high pressure homogenizer or the assistance of ultrasonic homogenizer. Our approach as a much more energy efficient method.

[0583] While our study focused was on developing BGI-based emulsion food products from Euglena biomass, the investigation of the preparation of BGI emulsions was carried out on the preparation methods, effects of the emulsion formulation and storage temperatures on the emulsion appearance and phase/micelle stability. The BGI emulsions have been successfully prepared from Canola oil, Palm oil, Coconut oil, Sunflower oil and Linseed oil when the BGI was premixed with the oils. Differing from reported methods, we first incubated BGI in Oil, and then homogenized it with water to give the BGI emulsion using a low power homogenizer. It was found that the inoculation time of BGI in oil was essential to the formation of BGI emulsions. Emulsions with different textures such as milk-like, cream like, yogurt-like and cheese-like were obtained by varying the BGI and oil contents in the emulsions. The emulsion can contain approximately 0.6% to 40% BGI and approximately 20% to 90% water, and 5% to 60% oil, causing the textures of emulsion to vary from milk-like, cream-like, yogurt-like to cheese-like.

[0584] In terms of emulsion phase separation (mainly gravitational separation), the milk-like texture emulsion had gravitational separation within 20 mins; 1-5% cream-like texture emulsion phase had separation within 4 hours, however, the emulsion with thicker than 10% cream and had phase stability at least 24 hours. Among the emulsions with different oils, sunflower oil based emulsions seemed to have best phase stability. It should be noted that the emulsion was reformed after a phase separation and the reformed emulsion had phase stability of more than 2 weeks, which may suggest that it gave better phase stability when the emulsion was prepared using a pulse homogenization. Additionally, it was found that the emulsion micelle and its size were stable after 4 weeks and should be noted that the Paramylon granules efficiently prevented the phenomena of flocculation and coalescence, making the emulsions stable.

[0585] In terms of the color, the higher the BGI content, the whiter the emulsion appearance, with a coconut oil base emulsion being whiter than the others.

|0586] In terms of thickness, the higher the BGI and the oil content, the thicker the emulsion was. Under this study, it was noted that the Linseed oil and Sunflower oil emulsions were thicker than others when the emulsion was freshly prepared, however, Coconut oil-based emulsion became the thickest over time.

[0587] In terms of the emulsion micelle size, it increased with the increase of the oil content and/or decrease of BGI content. The micelle size was less even with the average size decreasing when the BGI/Oil ratio decreased. In this study, the Coconut oil-based emulsions had a larger micelle size than others.

[0588] The effect of the storage temperature was investigated, and it was found that Emulsion micelles were broken after storage at -20°C overnight, but were stable at high temperatures (even at 80 °C for 2 hours).

[0589] It is interesting that we also notice that Coconut oil base emulsion gave the hardest texture after it was dried, but the hardness of Canola oil and Palm oil based emulsions were similar.

[0590] Similar to MCC (Micro Crystal Cellulose) Pickering emulsions, BGI emulsions have the potential for encapsulation of lipophilic compounds to be used in the food, cosmetic and pharmaceutical industries. The success in the formation of Euglena biomass based emulsions confirmed the possibility of encapsulation of nutrients. BGI can be used as a binder and filler, reinforcing agent or a viscosity regulator, and in food industries, BGI can be an emulsifier in pastes, creams, or sausages. Further, BGI in emulsion is not only an emulsifier, but also encapsulated in emulsion micelles as nutrients.

[0591] Potential Food Applications

[0592] BGI-oil-water emulsions have the potential for various applications in dairy analogues, spreads, dips and sauces. These emulsions may be used in the formulation of non dairy creamers, yogurt, cream cheese, cheese spreads, and cheese analogues, among other applications. The emulsions may act as a base for other ingredients to be added to improve texture, flavour and/or functionality. Or the emulsions products could stand alone and only need flavouring added.

Example 12: Additional Investigations of Beta Glucan Based Dairy Analogues [05931 Objective:

[0594] Major objective of this project was to develop dairy analogues such as creamer, cream cheese and yogurt using wet or dry beta glucan isolate (BGI) or its derivatives.

[0595] Key Summary

|0596] Creamer formulations were developed using both wet and dry BGI

[0597] Cream cheese formulations were developed using wet/dry BGI and RTG

[0598] Yogurt formulations were developed using wet/dry BGI and wet RTG

[0599] Beta Glucan Isolate Based Creamer

[0600] Objective:

[0601] Objective of this part of the project was to obtain a creamer formula using BGI which is stable during the storage in a refrigerator, and when added to hot coffee.

[0602] Background:

[0603] A creamer is an oil in water emulsion. An emulsion is a thermodynamically unstable system and the kinetic stability of the emulsion can be achieved by changing the composition of oil, water or emulsifiers in the systems. The creaminess of the creamer can be varied by adjusting the oil content of the emulsion, with increase in oil content, the creaminess increases. Emulsifiers are widely used to improve the stability of the emulsions. Monoglyceride based emulsifiers, proteins, lecithin etc. are commonly used as emulsifiers in creamers. For vegan creamer application, plant based emulsifiers are commonly used. The method/equipment used for emulsion preparation also determines the stability of emulsions, equipment which could enable the formation of small oil droplets is required to make stable emulsions. 39 different creamer formulations were investigated where different combinations the of ingredients and varying ratios of the following ingredients: Euglena derived BGI, Euglena derived Milled BGI, Euglena protein concentrate {Euglena protein rich flour, Euglena biomass, Euglena protein isolate could also be used), coconut oil, canola oil, sunflower lecithin, Pectin (pre hydrated), Oat milk powder, pea protein isolate (concentrate can also be used), Carrageenan, sunflower oil, gellan gum, xanthan gum, tricalcium phosphate, sugar, Myverol 18-92K, oat milk flour, Admul Ma 60-04K, Myverol 18-06pK, Myvacet 9-45K, guar gum (pre hydrated), guar gum (8/24), Xanthan gum+konjack gum, Myvacet, gum acacia+gellan gum and water.

[0604] Use of BGI as an emulsifier in creamer: It was found that oil-in-water emulsions can be prepared by using BGI as emulsifiers. To obtain these emulsions, different concentrations of BGI were added to vegetable oil and added this mixture to desired amounts of water and homogenized using a polytron blender (omni GLH polytron mixer at speed 9 for 5 minutes). However, when these emulsions were added to hot water/hot coffee, the BGI sedimented quickly and the oil came on the top of the coffee and formed a thick cream layer. It shows that the current formulation with BGI alone cannot be used as an emulsion stabilizer in a creamer unless further investigated. However, the white colour of the BGI can be utilized as whitener in a creamer and it can also add the immune boosting effect of BGI to the creamer. Therefore, more formulations were used to prepare emulsions by incorporating lecithin, mono diglyceride based emulsifiers, proteins and also tried using kitchen aid hand mixer, polytron blender and high pressure homogenization for emulsification.

[0605] BGI Creamer Stability and the Equipment Used for Emulsification

[0606] Emulsifiers used: Euglena protein concentrate, oat milk flour, pea protein, mono/diglycerides, and sunflower lecithin.

[0607] Equipment used for emulsification: KitchenAid 5-Speed Ultra Power Hand Mixer in Empire with only one wire whipper attachment and omni GLH polytron mixer and high pressure homogenizer (SPX RANNIE/GAULIN homogenizer-2 litre)

[0608] Method of Emulsion/creamer preparation: All water soluble ingredients (proteins, sunflower lecithin, hydrocolloids) were mixed with water and left under mixing for 2 hours for proper hydration. Coconut oil and canola oil were mixed (heated above the melting point of coconut oil to ensure proper blending) and BGI was added to the oil blend and also left the mixture stirring for 2 hours. The oil blend was then mixed with a water-emulsifier mixture and emulsified using either KitchenAid 5-Speed Ultra Power Hand Mixer in Empire with only one wire whipper attachment for 5 minutes or omni GLH polytron mixer at speed 9 for five minutes or using a high pressure homogenizer at 10,000 psi for 5 minutes.

[0609] The Euglena protein concentrate can make up about 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 18, 20, 25, 30, 35, 40, 45, 50 % W/W or more of the ingredients or final product. Flour, such as oat flour, can make up about 1, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 50 % W/W or more of the ingredients or final product. A protein source, such as pea protein, can make up about 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50% W/W or more of the ingredients or final product. A lecithin, such as sunflower lecithin, can make up about 0.1, 0.5, 1, 2, 3, 4, 5, 8, 10, 15% W/W or more of the ingredients or final product. An emulsifier, such as diglycerides, can make up about 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, 18, 20, 25, 30, 35, 40, 45, 50 % WAV or more of the ingredients or final product.

[0610] Stability study of creamers: Creamers prepared were poured into 15 mL centrifuge tubes and left undisturbed in a refrigerator to see any phase separation. Also a 15 mL of the creamer was added to 200 ml hot, freshly prepared black coffee to see the stability of the creamer in coffee. The coffee was left undisturbed for 30 minutes after the addition of creamer at room temperature to see any destabilization of creamer in coffee overtime.

[0611] Results and Discussions: Formulations 2-7 were prepared using both KitchenAid 5 -Speed Ultra Power Hand Mixer in Empire with only one wire whipper attachment and omni GLH polytron mixer. Irrespective of the emulsifier used or hydrocolloid used, all the creamers prepared using a kitchen aid mixer destabilized faster when added to coffee, a cream layer formed on the top of the coffee and BGI sedimented at the bottom of the coffee within 10 minutes after the creamer addition to coffee (See FIG. 28A). On the other hand there was no thick cream layer formation on the top of the coffee was not observed when the polytron blender was used for creamer preparation. These results suggest that the reduction in oil droplet size obtained using the polytron blender helped to improve the creamer stability in coffee. However, when a high pressure homogenizer, (an equipment which could make oil droplets significantly small) was used to prepare the emulsion (dairy formula 7), the creamer broke down immediately by forming small microgels and settled within two minutes by leaving the coffee in the original colour as can be seen in FIG. 29B. Although the droplet sizes of creamers obtained using high pressure homogenization compared to the one obtained using polytron blender, and provided higher stability for the creamer against phase separation while storing in room, the creamer obtained using high pressure homogenizer destabilized quickly in hot coffee.

[0612] In order to make sure this effect of high pressure homogenization is applicable to all types of hydrocolloid-pea protein combination, guar gum (pre hydrated) and carrageenan were used with pea protein in creamer formula (creamer formula 8-10). Same results were observed when the creamer was added to coffee, irrespective of type of hydrocolloid used.

[0613] The reason for the formation of gel-like substances was found to be due to the structural changes happening to protein and polysaccharides during the high pressure homogenization. The pea protein might have changed its structure in such a way to quickly form complexes with carrageenan and form coacervates in coffee as the coffee pH (4.5 to 5) and temperature was optimum for coacervate formation. Therefore, use of a polytron blender was recommended and used for future creamer preparation.

[0614] Although the creamer stability increased by using a better equipment for emulsification, the formation of cream layer was thicker than the commercial creamers and there was BGI sedimentation. Therefore, more formulations were tried by varying the concentration and type of hydrocolloids, proteins and emulsifiers. [06151 Effect of protein type used on creamer stability: Three types of proteins were used for creamer preparation at a concentration level of 1-2% in the formula. Pea protein isolate, Euglena protein concentrate and oat milk powder was selected for the study (Formula 11-13). Among them, the creamers prepared using pea protein isolate were whiter in colour and displayed a thin cream layer when added to coffee. Additionally, the creamer prepared using Euglena protein concentrate displayed strong marine flavour. However, BGI sedimentation was not affected by any protein used.

[061 ] Effect of emulsifier type used on creamer stability: Previous studies showed that use of BGI, protein and sunflower lecithin alone cannot prevent the destabilization of creamer in coffee and cream layer formation on the top of coffee when added to coffee. The objective of this study was to see if the addition of monoglyceride based emulsifiers and addition of milled paramylon instead of paramylon can increase the stability of creamer.

[0617] Emulsion preparation: Creamers were prepared using ingredients shown in Table 27. Two different concentrations of 6 different emulsifiers were tried in the basic dairy formula given in Table 28.

Table 27. Basic dairy formula used for emulsion (creamer) preparation. Emulsifier concentration was varied from 0.8% to 1.6%

[0618] In order to make the emulsions, oil blends were prepared first and emulsifiers were added into the oil mixture and heated until the emulsifier was completely melted and dissolved in oil. All the other ingredients except BGI were dissolved in water and let mix for a minimum of 2 hours to get proper hydration for protein and hydrocolloids. Then the water mixture was mixed with an oil mixture and the temperature of the mixture was allowed to rise until the emulsifiers were dissolved well again in the mixture. (Emulsifiers with higher melting temperature were tuned into solid when water mixture at room temperature was poured into the oil-emulsifier blend, but it was required to make sure that the emulsifier was in liquid state during the preparation of emulsions). The mixture was then blended using an Omni GLH polytron mixer at speed 9 for 5 minutes to form the emulsion. The BGI was then added to the emulsion and blended again using an Omni GLH polytron mixer at speed 9 for 3 minutes to form the non-dairy creamer.

[0619] In order to see the stability of different creamers, 15 mL creamers were added into 200 mL hot coffee and mixed well. The mixture was monitored for 10 minutes to see if there as any formation of cream layer on top of coffee and any sedimentation at the bottom.

Table 28. List of mono/diglyceride based emulsifiers used for creamer preparation

[0620] Results and Discussions: Creamers were added to hot coffee and monitored for 30 minutes. Generally, the stability of creamers increased with addition of mono/diglyceride based emulsifiers. However, the stability and thickness of the cream layer formed was determined by the type of emulsifier used. Myverol 18-92K and Myverol 18-06K displayed the highest effectiveness, as there was no thick cream layer observed on the top of the coffee after 30 minutes of storage.

[0621] Conclusions/recommendation: The addition of mono/diglyceride emulsifiers in addition to sunflower lecithin can improve the stability of creamers made of BGI in hot coffee. Myverol 18-92K (1.0 g) and Myverol 18-06K (1.0 g) were found to have the emulsifiers displayed highest effectiveness. Use of Myverol 18- 92K will be better for future application since it had a lower melting temperature than Myverol 18-06K ( 1.0 g).

[0622] Although the cream layer formation on the top of the coffee was eliminated by using the emulsifiers, the BGI sedimentation was still occurring. The best way might be using some hydrocolloids in the creamer formulations to prevent or reduce the BGI sedimentation

[0623] Effect of hydrocolloid type used on creamer stability: Hydrocolloids such as carrageenan, guar gum, xanthan gum, locust bean gum, gellan gum etc. are molecules capable of interacting with other hydrocolloids, proteins and are able to modify the viscosity, gelling, thickness, sedimentation and phase separation of mixers involving these hydrocolloids. This property of gums/hydrocolloids could be utilized to reduce the sedimentation of BGI.

[0624] Different concentrations of hydrocolloids listed in Table 29 were used for creamer formulations, using the formula shown in Table 31. Protein, hydrocolloid and sunflower lecithin were dissolved in water and left it mixing for 2 hours. The oil blend was prepared with the emulsifier and mixed with water-protein and hydrocolloid mix. The mixture was then homogenized with a polytron blender for 5 minutes and then BGI was added to emulsion and mixed again using the same polytron for 1 minutes. The creamers obtained were added to coffee (15 mL of creamer in 200 mL of coffee).

Table 29. Different hydrocolloids and their concentrations used for emulsion preparation

Table 30. Summary of performance of creamer (phase stability by itself and in hot coffee)

[0625] This study showed that the carrageenan is a good hydrocolloid, because it which prevented/reduced the BGI sedimentation. However, it can forming a self standing gel.

[0626] In order to figure out which ingredient was responsible for the gelation of creamer emulsion in the refrigerator, five different creamers were prepared similar way as the control (Table 31) creamer but without (1) coconut oil (coconut oil replaced with sunflower oil), (2) without sunflower lecithin, (3) without sugar, (4) without carrageenan and (5) without BGI. These creamers were also left at the refrigerator for 2 days and observed the changes. The following were observed (See FIG. 30).

[0627] All creamers except the one prepared without carrageenan were turned into self standing gels. The creamer without carrageenan was pourable as it was on the day it was prepared, indicating that carrageenan was responsible for the gelation of creamer.

[0628] However, when this creamer was poured into coffee, the BGI was sedimenting within 2 minutes after addition, while no cream layer formation on top of coffee was observed for 30 minutes. This indicates that some hydrocolloids if not carrageenan was required to prevent or reduce the sedimentation of BGI.

[0629] All these gels were melted while heating, and reformed gels while cooling.

[0630] All these gels break down to thick fluid while mechanical forces are applied like stirring/blending but reforms gels/ thicken when stored at the refrigerator.

[0631] Gel strengths were different for creamers: Gel without sunflower lecithin was least strong. It broke while taking out from the centrifuge tube and displayed a yogurt like consistency. Control creamer, creamer without coconut oil and without sugar displayed pretty much the same strength. The strength of the creamer without BGI was not as strong as control, but was stronger than the one without sunflower lecithin.

Table 31. The basic formula used for creamer preparation with different hydrocolloids

[0632] This study showed that, although carrageenan was able to prevent/reduce BGI sedimentation, due to the gelling of carrageenan emulsion in the refrigerator this hydrocolloid is not best for creamer preparation. However, due to the melting and gelling ability of these emulsion, it is possible to use this emulsion base for future non-dairy yogurt, cheese, and cream cheese applications

[0633] BGI Based Creamers without any Protein: More studies were conducted in- order to see if the creamer formulations can be obtained without any protein. Carrageenan, guar gum (8/24), gum acacia+gellan gum were explored for this purpose using the formulas 34-38.

[0634] Results and discussions: Without protein, the carrageenan containing emulsions were forming gels at the refrigerator. But no gel formation was observed in emulsions containing guar gum. When this creamer was added to coffee, although there was BGI sedimentation, it was much less than when there was no gum present. However, when the acacia gum+gellan gum was used, the creamer sedimented in coffee much quicker than BGI alone and the coffee became dark overtime. See FIG. 31.

[0635] Use of BGI slurry and Milled BGI as a Replacement for Spray dried BGI

Powder

[0636] BGI Slurry: Use of BGI slurry obtained from the co-purification process before spray drying for creamer application was investigated for creamer preparation. For this purpose, the BGI slurry was diluted to 3% BGI (3% solid) and in 52 g of this slurry, selected gum (guar gum (8/24)), sunflower lecithin and sugar was dissolved and left the mixture under stirring for a minimum of 2 hours. Canola oil was then blended with myverol 18-92K (mono/diglyceride emulsifier) and the oil blend was then mixed with BGI slurry and homogenized the mixture using a polytron blender for 5 minutes at speed 9. The creamer obtained was then added to coffee to see the stability of creamer in coffee. Remaining creamer was stored in a refrigerator to see the stability of the creamer.

[0637] Results and discussions: The creamer obtained using BGI performed similarly like the corresponding creamer obtained using BGI powder, except there was slightly higher BGI precipitation rate was observed when BGI slurry based creamer was added to coffee.

[0638] Milled paramylon: Use of milled paramylon for creamer application was explored by replacing BGI in the creamer formula with milled paramylon. Milled paramylon obtained was a gel like wet mixture with 20% solid. It was diluted to provide 3% BGI in the slurry and prepared emulsion using this slurry (Formula 34-35) in the same way explained before. The creamer obtained was added to coffee and also stored in the refrigerator to see stability. It was expected that since the milled paramylon had smaller granules than spray dried BGI, the BGI sedimentation rate was low when milled paramylon was used for creamer preparation.

[0639] Results and discussions: The creamer obtained looked similar to powder BGI based creamer. However, when the creamer was added to coffee, the small gel like particles were formed and sedimented quickly. The milled paramylon was precipitated within 15 minutes while paramylon (BGI) was still dispersed well in the coffee. Reformation of gel like particles and formation of larger particles leads to the sedimentation in coffee; however, the reason behind the gel formation of milled paramylon in hot water/coffee is still undetermined.

[0640] Major Conclusions from the study:

[0641] Non-dairy creamer was obtained using wet or dry BGI as a whitener.

[0642] A hydrocolloid/gum was required to prevent the sedimentation of BGI when the creamer was added to coffee.

[0643] The ideal hydrocolloid used for creamer application did not increase the viscosity of the creamer during refrigeration did not prevent or reduce BGI sedimentation while added to coffee.

[0644] Guar gum (8/24) was found to be the best gum that could be used for BGI based creamer.

[0645] The best formula for BGI based creamer is shown in Table 32, however variations in ingredients and amounts of ingredients are contemplated. In an embodiment BGI is present at about 1, 2, 3, 4, 5 % W/W or more, the water is present at about 30, 40, 50, 60, 70% W/W or more, the oil is present at about 2, 3, 4, 5, 6 % W/W or more, the lecithin is present at about 0.2, 0.3, 0.4, 0.5, 0.6 % W/W or more, the distilled monoglyceride prepared from vegetable oils and fats is present at about 0.2, 0.3, 0.4, 0.5, 0.6. 0.7, 0.8, 0.9, 1, 2, 3, % W/W or more, the hydrocolloid (e.g., guar gum)is present at about 0.001, 0.01, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6 % W/W or more, the sugar is present at about 0.5, 1.0, 2.0, 3.0, 4.0, 5.0% W/W or more.

Table 32. The best BGI based creamer formula.

[0646] BGI Based Yogurt

[0647] Background:

[0648] The emulsions prepared using pea protein-carrageenan and BGI were found to form a self standing gel while stored in the refrigerator, and have a huge potential to be used as yogurt cheese, cream cheese, etc.

[0649] Effect of Protein and Carrageenan Content and the Ratio of Protein Carrageenan Content on the Gel strength of the Emulsions for Yogurt Applications

[0650] Gel strength of gelled emulsions may vary with protein, carrageenan and fat content of the emulsions. A detailed study was conducted by varying the concentration of carrageenan from 0.25 to 0.82% by fixing the protein concentration at 2.5% and varied protein content from 2.5% to 10%.

[0651] Method of Emulsion Preparations:

[0652] Emulsions were prepared using the dairy formula 1-5 (Table 33). The protein, carrageenan and sunflower lecithin was dissolved in water and left the mixture under stirring for 2 hours for proper hydration at pH 7. The oil blends were prepared and the emulsifier was added above the melting point of the Myverol emulsifier. Then the oil blend was added to the protein solution and homogenized for 5 minutes using the polytron blender at speed 9. Required amount of BGI powder was then added to the emulsion and homogenized using the same polytron blender for 1 minute at speed 9. The emulsions were then spilt into half and the pH of half of the emulsion was changed to pH 4.5. All the emulsions at pH 7 and pH 4.5 were stored in the refrigerator overnight and analyzed.

Table 33. BGI based yogurt formulas (1-7). Milled paramylon used was 20% solid, it was diluted to obtain 1.5 g dry BGI in 52 g total.

[0653] Results and Discussions: All emulsions prepared with different protein and carrageenan concentrations formed a gel at pH 7. Gel strength increased with both protein and carrageenan content at pH 7. The white colour of emulsions reduced when the pH of the emulsions were reduced to pH 4.5. The gel strength of the pH 4.5 emulsions were higher than corresponding pH 7 emulsions when the protein to carrageenan ration was 6 to 12. These optimum ratios of protein and carrageenan can be utilized for yogurt application. But when this ratio was higher, weak gels were formed at pH 4.5.

[0654] At higher protein (P=7) and carrageenan concentrations (C=0.5), even before emulsification the protein-carrageenan blends formed thick gels, which made the emulsification process difficult. Therefore, a concentration of carrageenan between 0.5-0.8 was preferred.

[0655] Conclusions: Gel strength of emulsions were increased with both protein and carrageenan concentration at pH 7

[0656] Optimum ratio of protein and carrageenan was required to obtain sufficient gel strength for yogurt application at pH 4-5

[0657] Effect of pH of the Protein Solution on the gel strength of the Emulsion for Yogurt Application

[0658] The emulsions were prepared using protein-carrageenan solution at pH 7. However, the targeted products had a pH around 4-5. There were two ways that pH was achieved: emulsion was prepared using a protein-carrageenan solution at pH 7 and the pH of the emulsion was changed to pH 4-5; or the protein-carrageenan solution was prepared at pH 4-5 and used for emulsion preparation. However, if there was any difference between these emulsions, if one was more suitable for the targeted application, it was unknown. Also, there were different ways we incorporated BGI into these emulsions: either BGI powder or slurry was added after the emulsion was prepared, or BGI powder/slurry was added to the protein- carrageenan solution before emulsification, or BGI powder was added to oil before emulsification. It was unknown if the way BGI was added to emulsion affected the product. The objective of the current study was to see if the pH of protein-carrageenan solution, or the way BGI incorporated into the emulsion, would affect the final emulsion properties.

[0659] Method of emulsion preparation:

[0660] The emulsions were prepared using the basic formula given in Table 34. Table 34. Basic formula used for emulsion preparation for yogurt application

[0661] In trial 1 and 2, protein, carrageenan and sunflower lecithin solution was prepared in 50 mL water at pH 7 and pH 4.5, respectively and left at room temperature for 2 hours to obtain proper hydration. Both oils were mixed and Myverol 18-92K emulsifier was added to the oil blend. This blend was then added to the protein solution and homogenized for 5 minutes using a polytron at speed 9 to obtain emulsion. BGI was then added to these emulsions and blended using the same polytron at speed 9 for 1 minute. The pH of the pH 7 emulsion was then adjusted to pH 4.5 using 4M HC1 and compared with emulsion prepared using protein solution at pH 4.5. In trial 3, the BGI powder was added to the oil blend, and used this blend for emulsion preparation. In trial 4 and 5, the BGI powder and BGI slurry, respectively, was added to protein, carrageenan and sunflower lecithin solution at pH7. The emulsions were then prepared by adding oil blend to this solution using the same conditions explained in trial 1 and 2.

[0662] Results and discussions: Emulsions were prepared as follows: Emulsion prepared using protein-carrageenan solution at pH 4.5, emulsions prepared using protein- carrageenan solution at pH 7, emulsion prepared using protein-carrageenan solution at pH 7 which was then reduced to pH 4.5, emulsion prepared using BGI powder dissolved in oil, emulsion prepared using BGI powder mixed in protein-carrageenan solution, and the emulsion prepared using BGI slurry mixed in protein-carrageenan solution.

[0663] The emulsion prepared using protein-carrageenan solution at pH 7 and pH 4.5 appeared completely different, the emulsion at pH 7 formed a self standing gel while storing in the refrigerator while the emulsion at pH 4.5 did not form a self standing gel. The emulsion at pH 7 was whiter in color. When the pH of the emulsion at pH 7 was reduced to pH 4.5, a curd-like structure was formed, which was also not looking like the emulsion prepared using protein-carrageenan solution at pH 4.5. [0664] There was no difference between the emulsions formed using trial 3, 4 and 5. It was easier to dissolve BGI slurry into the protein solution than the powder. It tells that it doesn't matter at what stage or if it is in the oil or in the water phase the BGI was added, the emulsion will be the same for this creamer example.

|0665] Conclusions:

[0666] For yogurt or cheese or cream cheese like application, it was better to prepare the protein-carrageenan solution at pH 7, and to reduce the pH after emulsification. It as also possible to use some cultures to reduce the pH if the emulsion was prepared at pH 7 (further optimization was required for that procedure). Since there was no difference between the way BGI was added, it was recommended to add BGI in protein-carrageenan solution, especially when BGI slurry was used, as it was easy to mix BGI slurry in water phase in this creamer example.

[0667] Use of wet/dry RTG as a replacer for BGI in Yogurt Application

[0668] The use of ready to gel beta glucan isolate (RTG) in wet form as a replacement for BGI in emulsions for yogurt application was investigated. It was expected that the gel strength of the emulsion will be increased with the addition of wet RTG

[0669] Method of emulsion preparation: The wet RTG was mixed with enough water to make 3% BGI in 52 g mixture. The wet RTG used in this study was washed three times with 8% solid. Protein, carrageenan, and sunflower lecithin were mixed with RTG slurry. The oil blend with emulsifier was also prepared and added to the water blend and homogenized the mixture using a polytron at speed 9 for 5 minutes to form the emulsions.

[0670] Results and Discussions: The image of emulsion prepared using wet RTG is displayed in FIG. 32. The emulsion prepared using wet RTG was thicker gel like right after preparation unlike the one prepared using BGI, which only formed gel like structure after refrigeration. Refrigeration of the emulsion prepared using wet RTG increased its gel strength further.

[0671] Use of Milled Paramylon in Emulsions for Yogurt Uike Application

[0672] Milled paramylon as a replacer for BGI was investigated. Instead of BGI powder, an equivalent amount of milled paramylon was used in the formulation (see Formula 7 in Table 33). Milled paramylon was diluted in water to obtain an equivalent amount of 3% of BGI. Protein, carrageenan and sunflower lecithin were dissolved in 52 g milled paramylon slurry and after two hours it was mixed with oil blend with mono/diglyceride emulsifier and the mixture was then homogenized to obtain the emulsion using a polytron blender for 5 minutes at speed 9. The emulsions were prepared at pH 7 and later reduced the pH to 4.5. Both emulsions were stored in the refrigerator. A BGI powder based emulsion was also prepared and stored as a control. After storing in the refrigerator for a day the emulsions at both pH were evaluated. There was no difference between BGI or milled paramylon at both pH values.

[0673] BGI Based Cream Cheese

|0674] Objective: Objective of this study was to form a non-dairy cream cheese formulation using ready to gel beta glucan isolate (RTG) or beta glucan isolate (BGI) as a base. The target was to achieve at least 2 g protein per 30 gram serving and less than 17% fat per serving with a similar appearance and texture as that of non-dairy cream cheese or real cream cheese.

[0675] Materials: RTG, BGI, hydrocolloids (pre hydrated pectin, carrageenan, xanthan gum), water, sunflower lecithin, oat milk powder, pea protein, Euglena protein concentrate.

[0676] RTG Based Cream Cheese

[0677] RTG was used due to its thickening properties compared to BGI. It was found that a combination of RTG with pre hydrated pectin can provide the thickness of the cream cheese required.

[0678] RTG Based cream cheese preparation procedure: Several formulations were prepared and tested using RTG and different proteins (oat milk flour, pea protein isolate, Euglena protein concentrate). The formulations contained combinations of the following ingredients, including testing different ratios of ingredients: Ready To Gel (RTG) BGI, Oil (1 : 1 canola and coconut), sunflower lecithin, pectin (pre hydrated), Euglena protein concentrate ( Euglena protein rich flour, Euglena biomass, Euglena protein isolate could also be used), Oat milk powder, coconut oil, sunflower oil, pectin, xanthan gum and konjac gum, potato starch, tapioca starch, sodium alginate, baking powder, sugar, salt, lemon juice concentrate, flavourings and or maskers, gellan gum, lactic acid powder, lemon, pea protein isolate (pea protein concentrate could be used), carrageenan, BGI, emulsifier (Admul), Myverol 18-92K, and water.

[0679] Required amount of water was taken in a measuring cup. Desired amounts of sunflower lecithin and oat milk flour/pea protein was blended well with water. Required amounts of pectin and RTG were measured and mixed well in the powder form. This mixture was then added to the water mixture and blended well. Required amounts of flavours/maskers were then added to this mixture. Desired amount of coconut oil in the solid form was then added to the water base and mixed well to facilitate the formation of cream cheese. The blend was then refrigerated. [0680] Results and discussions: 11 different formulations were prepared. Among them, dairy formula 9 and dairy formula 11 were found as the best formulations for non-dairy cream cheese applications. These formulations can be seen in Table 35.

[0681] As can be seen, the only difference between dairy formula 9 and 11 was the source of protein: in Formula 9 it was oats and in dairy formula 11 it was from pea. The cream cheese prepared using dairy formula 9 was dark yellow coloured as can be seen in FIG. 33. This was due to the dark color of oat milk powder. Also, the coconut flavour was overpowering in Formula 9. Therefore pea protein with a lighter yellow colour and a higher amount of cheese flavours were used in Formula 11. In dairy formula 9, the protein per serving was about 0.45 g while it was about 2.4 g in dairy formula 11.

Table 35. Formulation 9 and 11 for RTG BGI cream cheese formulations.

[0682] Hardness and spreadability of dairy formula 9 and dairy formula 11 was very close to commercial cream cheese analogues. However, when tasted, both these products displayed a slimy mouthfeel and required more cheese flavouring.

[0683] In order to improve the mouthfeel of the cream cheese analogue, a different preparation method was used. In this method, coacervation of pea protein isolate with different hydrocolloids were utilized to improve the mouthfeel. The idea was, when the coacervates are formed between pea protein and the hydrocolloids were formed, they act similar to casein coagulation in real cream cheese. Moreover, different flavours will be used to improve the flavour of the cream cheese to make it a cream cheese spread.

[0684] Procedure 2:

[0685] In this procedure only pea protein was utilized as there were no large coacervate formation between protein in the oat milk flour and xanthan gum/gellan gum (hydrocolloids used in this recipe) was found. The ingredients and its concentrations used in this formulation is shown in Table 36.

[0686] In this procedure, pea protein isolate, xanthan gum and gellan gums were dissolved in water first. The pH of the solution was around pH 7 (pH of pea protein isolate was pH 7). Then lemon concentrate was added to the protein-gum solution until the pH of the solution was around pH 3.5 to 4.5. The mixture was then gently stirred, large coacervates of pea protein and hydrocolloids were formed and started separating from the water phase. At this point, all the other ingredients were added to the mixture and mixed well. Sriracha and lime were added to improve the flavour of the cream cheese

Table 36. Formulation of cream cheese developed using complex coacervation

[0687] Results and discussions: As expected, the slimy mouthfeel was reduced in this product. Sensory evaluation of the product showed that the texture of the product was much less slimy but it was less smooth and slightly gritty. Also it was bit saltier. The grittiness was due to the improper hydration of pea protein. In order to avoid this, pea protein and hydrocolloids must be hydrated 2 to 3 hours in water prior to adding any other ingredients. Also less amount of Sriracha needs to be added to make it not as salty.

[0688] Conclusions: A cream cheese analogue can be obtained using RTG as a gelling agent with pea or oat milk flour as a protein source. Coacervate formation of proteins and hydrocolloids prior to the addition of other ingredients for the cream cheese formation can reduce the slimy mouthfeel and improve the texture of the product. Any protein source with a lighter colour than pea protein with high protein content can be used as an alternative protein source in the cream cheese formulation.

[0689] BGI Based Cream Cheese: Cream cheese formulations were developed using BGI and carrageenan. The formulations (Formula 12-18) were used.

[0690] Method of preparation: Protein and other water soluble ingredients were dissolved in water and left for proper hydration. The oil-and mono/diglyceride emulsifiers blended above the melting point of the emulsifier and the oil blend was added to the water mixture and blended well using a Kitchen aid hand mixer.

[0691] Results and Discussions: A representative picture of cream cheese prepared using BGI is shown in FIG. 34. The cream cheese prepared using BGI was whiter in color than the one prepared using RTG. When the protein concentration was high, there was gritty mouthfeel due to improper hydration of pea protein. The water mixture formed a gel when the protein concentration was 15% or high. Below 12% of protein was preferred for cream cheese application. Also a gritty mouth feel was detected when Admul mono/diglyceride emulsifier was used after refrigeration. It was due to the crystallization of these emulsifiers. However, the cream cheese prepared with lower concentration of protein and myverol 18-92K was smooth and formed the right texture when stored in the refrigerator.

[0692] The cream cheese with BGI was prepared two different ways, first prepared the mixture at pH 7 and added either lemon juice or lactic acid to reduce the pH to pH4-5, or prepared the mixture at pH 4-5 by changing the pH of the protein solution to pH 4-5 (using lemon juice or lactic acid) before mixing everything else. When the pH of the protein solution was at pH 4-5, the viscosity of the protein-carrageenan blend was not highly viscous. However, the cream cheese developed using the mixture at pH7 and reduced the pH later to 4.5 was better than the cream cheese developed using the protein mixture at pH4-5.

[0693] Even with the cheese flavours used in these formulations, right cream cheese flavour was not obtained to mimic real cream cheese. However, the flavour was enhanced by using herbs such as parsley, garlic, oregano or thyme and the flavour was comparable with non-dairy cream cheese found in the market. [0694] Conclusions/recommendations: Cream cheese like products can be prepared using both RTG and BGI. The cream cheese had the right texture when the mixture was water- protein at pH 4-5 when using RTG while it was better to be at pH7 when using BGI and reduced the pH to 4-5 later.

|0695] Overall conclusions of the Project: Dairy analogues can be prepared using BGI powder or slurry and BGI derivatives (RTG, milled paramylon)

[0696] Guar gum (8/24) was found to be the best gum that can be used for BGI based creamer

[0697] Yogurt like emulsions can be prepared using BGI, RTG (wet) or milled paramylon

[0698] Texture of the yogurt/cream cheese like emulsion was right when the emulsion was prepared at pH 7 first and reduced the pH later t pH 4-5 when BGI was used in the formula

[0699] Future study required

[0700] Although formulations were developed for creamer, yogurt and cream cheese analogues using BGI and its derivatives, the shelf life of these products were not done yet. Stability of these products against microbial spoilage, and best storage conditions and preservatives etc. needs to be investigated.

Example 13 - Euglena Meat Analogue Processes and Formulations

[0701] Formulations were developed for both low moisture textured vegetable protein (TVP) and high moisture extrusion cooking (HMEC) extruded products using dry Euglena- derived protein rich flour and Euglena- derived beta-glucan isolate (Table 37, Table 38, Table 39, Table 40). Examples of TVP and HMEC based Euglena meat analogues are shown in FIG. 35. The Euglena protein rich flour in combination with pea protein at the two different moisture conditions formed meat like textured product. The TVP Euglena formulations were able to absorb an amount of water of about 2.5 times their weight. These products were also used to prepare soups, tuna wraps, bacon analogues, pulled pork and crisps as non-limiting examples. Examples of HMEC process Euglena meat analogues in a bacon like product and a pulled pork product are shown in FIG. 36.

[0702] Eight different formulations were developed and tested under HMEC extrusion conditions as shown in Table 37 and Table 38. Euglena derived flour was added in the proportions indicated in the tables, in addition to other ingredients such as pea protein isolate, peal hull fiber, com starch, masker, wheat gluten, Euglena beta-glucan isolate and water. HMEC Formula 5 (Table 38) performed the best in the HMEC process. The dry basis of HMEC formulations 1-4 is shown in Table 39.

Table 37. High Moisture Extrusion Formulations 1-4

Table 38. High Moisture Extrusion Formulations 5-8

Table 39. High Moisture Extrusion Formulations 1-4 Dry Basis

Table 40. Low Moisture Euglena Based Extruded Product Formulation

Example 14 - Euglena Textured Vegetable Protein Soup Formulations

[0703| Purpose: To create dry soup mixes that highlight our new Euglena (ETVP) and as proof of concept of ETVP as a chicken replacement.

[0704] HMEC Formula 6 (12% pea protein concentrate, 49.5% Euglena flour and 37.5% pea protein isolate) was chosen for the soups. HMEC Formula 6 was the only one that did not contain any maskers. Unfortunately, the maskers chosen in these formulations gave all other formulas a sweet aroma and taste which was not ideal for this application. However, the maskers can be replaced with a more savory or neutral masker. Vegan Chicken and Noodle Soup (FIG. 37A) and Chicken Coconut Thai Soup (FIG. 37B) were made (Table 41 and Table 42).

[0705] There was no deterioration of the ETVP pieces during the boiling process of the soup mixes. All ingredients were bought at the Bulkbam, Peterborough Ontario, Canada. The ETVP was an effective chicken alternative in this application. To create, all ingredients were mixed together and placed in packaging or layered in packaging.

Table 41. Chicken Coconut Thai Soup

Table 42. Vegan Chicken and Noodle Soup

Example 15 - Euglena Textured Vegetable Protein Tuna Formulations

[0706] A formulation using ETVP was developed as seafood alternative (Table 43, FIG. 38). HMEC Formula 6 (12% pea protein concentrate, 49.5% Euglena flour and 37.5% pea protein isolate) was chosen for the tuna. HMEC Formula 6 was the only one that did not contain any maskers. Maskers were found to give a sweet aroma and taste which was not ideal for this application. [0707] The small extruded pieces were used for the tuna as it was already the perfect size and shape to mock canned tuna.

Table 43. Flaked Tuna Formulation

[0708] All ingredients were mixed together in a large bowl until ETVP was hydrated. ETPV was spread in a thin layer on a parchment paper lined dehydrator tray and dehydrated at 52°C for 4 hours or until ETVP was dehydrated again.

[0709] Rehydrating instructions: ingredients were mixed well and were allowed to sit for 2 minutes, stirring occasionally. Rehydrated ETPV was used in place of regulate canned tuna.

[0710] Sensory Profile: Mild fishy notes came from the seaweed and natural Euglena flavour. Smoky, more of a smoked trout then a tuna flavour, umami, no Euglena flavour.

[0711] Mouthfeel: Similar to canned tuna, flaky. Aroma: smoky, fish notes. Appearance: A little brown compared to regular canned tuna. Similar in shape and size of regular canned tuna.

Example 16 - CHICKEN NUGGET, a Euglena TVP based breaded chicken nugget

[0712] In this study, a combination of Water, Euglena TVP (Pea Protein, Euglena, Natural Flavor), Panko Breadcrumbs (Wheat Flour, Cane Sugar, Yeast, Salt), Canola Oil, Mung Bean Protein, Wheat Flour, Yeast Extract, Methylcellulose, Coconut Oil, Sea Salt, Commeal, Dehydrated Garlic and Onion, Spices, Sugar, Smoked Paprika, Natural Flavor, Konjac Gum, Chickpea Flour, Rice Starch was used to create the final chicken nugget food product.

[0713] Methods and Materials: The formulation that was used in this study for a chicken nugget included Euglena TVP at approximately 20% of the overall recipe (Table 44). [0714] Base: For this analogue the ingredients were combined with water, mixed and set aside to hydrate and then chilled in a cooler for 30 minutes. After chilling for 30 minutes, the mixture was removed from the cooler and pulsed in a food process until “Fine grind”, (similar to that of a non-Panko bread crumb), particle size was achieved. Canola oil, an antioxidant (Dadex AR HS), and slightly warmed coconut oil were combined in a kitchen aid stand mixer and mixed together with the above chilled ingredients using a paddle mixer until dispersed, about 1 minute. Mung bean protein and methylcellulose were added to the mixture in kitchen aid, and mixed for 1 minute. During this time, the side of the bowl were scraped down with a spatula. Water was added and the ingredients were mixed for 2 minutes until mixture became sticky and cohesive. The mixture was then formed into 15g pieces and put into a cooler or freezer to chill until set.

[0715] Wet Batter and Coating: The remaining ingredients were then combined and whisked together, and set in a cooler to chill for 30 minutes, whisking again prior to use to ensure the flour and starch remained in the solution. The chilled and formed bases were then dipped into the batter, allowing excess batter to drip off, then the formed base was coated in the breading mixture, the coating was firmly pressed into the nugget surface to ensure it was evenly coated. Once complete, the breaded bases were immediately placed back into the freezer to maintain and set the shape. The finished product was covered, sealed, and stored in the freezer.

[0716] To heat for consumption, an oven was preheated to 350 °F. Nuggets were then placed on a tray and heated for 14-16 minutes until the internal temperature reached 165 °F. The nuggets were turned once, halfway through heating to ensure cooked through. FIG. 40 is an example of the finished product.

Table 44. Euglena TVP Chicken Nugget Formulation

Example 17 - Euglena TVP Sausage Link

[0717] In this study, a combination of Water, Euglena TVP (Pea Protein, Euglena, Natural Flavor), Coconut Oil, Canola Oil, Pea Protein, Seasoning (Dextrose, Maltodextrin, Sea Salt, Onion Powder, Yeast Extract, Garlic Powder, Spices, Caramelized Sugar, Natural Flavor, Canola Oil, Grill Flavor), Methylcellulose, Mung Bean Protein, Sea Salt, Citrus Fiber, Black Malt Powder, and Konjac Gum was used to create the TVP Sausage link.

[0718] Methods and Materials: In this study, the formulation used to create the sausage link is listed in (Table 45), which includes approximately 10% of Euglena TVP. Ingredients were placed into a food processer and broken down until ground down to very fine granules, similar to breadcrumbs. Once finely ground, the ingredients were combined with the TVP and allowed to rest in a cooler for 30 minutes. After 30 minutes, the mixture was removed from the cooler and combined with seasoning, in a mixer, mixing on low for 5 minutes. Mixture was again chilled and stuffed into casing in 22-24 gram links, in 19 mm casings.

[0719] To prepare the final sausage link product, frozen links were placed in a non stick skillet and cooked over medium heat for 10-12 minutes, turning frequently to brown evenly. Sausage links were cooked through when an internal temperature of 165 °F was reached. FIG. 41 is an example of the finished product.

Table 45. Euglena TVP Sausage Formulation

Example 18 - Euglena TVP Pulled Pork [0720] In this study, a combination of BBQ Sauce (Tomato Puree (Water, Tomato Paste), Brown Sugar, Molasses, Distilled Vinegar, Water, Com Starch, Salt, Spices (including Black Pepper, Paprika, Chili Pepper), Onion, Garlic, Natural Flavor), Water, Euglena TVP (Pea Protein, Euglena, Natural Flavor), Coconut Oil, Yeast Extract, Dehydrated Garlic and Onion, and spices were used to create a final BBQ pulled pork analogue.

[0721] Methods and Materials: In this study, the formulation used to create the TVP Pulled pork is listed in (Table 46), which includes approximately 8% of Euglena TVP. The TVP was combined with water and allowed to hydrate well. Once hydrated, the TVP was heated to 320°F for approximately 5-10 minutes to drive off moisture. Once moisture was reduced, fat was added to the pan and the TVP was allowed to crisp up until it began to brown, approximately 5 minutes at 320°F. Once browned, barbeque sauce was added approximately one cup at a time and continued to cook down until barbecue sauce was partially absorbed, approximately 10 minutes at 180°F. Approximately half of the chosen barbeque sauce was reserved to finish with. The ETVP pulled pork can be consumed immediately, or optionally, frozen and thawed for use.

[0722] To consume, thaw in a refrigerator prior to use. To warm, cover with a paper towel and microwave for 1-2 minutes stirring halfway through the heating time. Alternatively, the pulled pork analogue can be placed in a non-stick skillet over medium heat for approximately 3-5 minutes, stirring frequently. FIG. 42 is an example of the finished product.

Table 46. Euglena TVP Barbecue Pulled Pork Formulation

Example 19 - Euglena TVP Fish Stick

[0723] In this study, a combination of Water, Euglena TVP (Pea Protein, Euglena, Natural Flavor), Canola Oil, Panko Breadcrumbs (Wheat Flour, Cane Sugar, Yeast, Salt), Breadcrumbs (Wheat Flour, Evaporated Cane Sugar, Sea Salt, Yeast), Mung Bean Protein, Yeast Extract, Methylcellulose, Natural Flavor (Yeast Extract, Sugar, Com Maltodextrin, Natural Flavor, Salt), Commeal, Coconut Oil, Dehydrated Garlic and Onion, Spices, Sea Salt, Chickpea Flour, Konjac Gum, Rice Starch, Granulated Sugar, Celery Powder, Smoked Paprika were used to create a Fish Stick analogue.

[0724] Methods and Materials: In this study, the formulation used to create the sausage link is listed in (Table 47), which includes approximately 20% of Euglena TVP.

[0725] TVP Base:

[0726] TVP and dry ingredients were mixed together, adding cold water to hydrate while mixing well. The mixture was allowed to chill in cooler for 30 minutes. After chilling for 30 minutes, the mixture was removed from cooler and pulsed in a food processor until a "fine grind" particle size was achieved. In a Kitchen Aid, the above chilled ingredients were mixed with the coconut oil, Dadex AR-HS, and canola oil using paddle mixer until dispersed, about 1 minute. Mung bean protein and methylcellulose were added to the mixture in kitchen aid for 1 minute. During this time, the sides of the bowl were scraped down with a spatula. Water was added and mixing continued for 2 minutes until mixture became sticky and cohesive. The mixture was formed into 15 g pieces (strips) and chilled in the cooler or freezer until semi -firm (10-15 min).

[0727] Wet Batter and Coating:

[0728] All remaining ingredients were combined, whisked together, and left to chill for 30 minutes. Prior to use, the mixture was whisked again as well as during use to ensure flour/starch stayed in solution. Ingredients were combined and set aside.

[0729] The chilled and formed bases were removed from freezer/cooler and dipped into the batter. Excess batter was allowed to drip off, then the formed bases were coated in the breading mixture. The coating was pressed firmly onto the nugget surface to ensure even coating and immediately placed back into freezer to set shape. Cover, seal and store frozen.

[0730] The fish sticks were heated for consumption, baked from frozen at 350°F for approximately 15-20 minutes. FIG. 43 is an example of the finished product. Table 47. Euglena TVP Fish Stick Formulation Example 20 - Italian Dinner Sausage with Euglena TVP

[0731] In this study, Water, Euglena TVP (Pea Protein, Euglena, Natural Flavor), Coconut Oil, Canola Oil, Seasoning (Spice, Sugar, Dried Garlic, Paprika, Herbs, Natural Flavor), Citrus Fiber, Methylcellulose, Mung Bean Protein, Konjac Gum, Sea Salt, Smoked Paprika, Natural Flavor, and Black Malt Powder were combined to create an Italian Dinner Sausage.

[0732] Methods and Materials: In this study, the formulation used to create the sausage link was listed in (Table 48), which included approximately 15% of Euglena TVP. TVP was broken down in the food processor to fine granules, similar to breadcrumbs. Boiling water and Citri-Fi TX10 were combined with TVP, mixed well and chilled for 30 minutes. Mung bean protein, konjac, Italian seasoning, sea salt, paprika, Ultra Seal 160 Meat (juicy) Flavor, and black malt were combined with TVP in themixer and mixed on low 1 minute to combine. Citri-Fi 100FG was dissolved into liquid coconut oil and then water was added. The mixture was emulsified with stick blender and allowed to chill for 24 hours. Then we combined emulsified cooled fat with TVP mixture in a stand mixer with paddle attachment. It was mixed for 2 minutes, until thoroughly dispersed. Then we added canola oil to the TVP mixture in the stand mixer, mixed for 1 minute until thoroughly dispersed. The process was repeated with methylcellulose and water. It was chilled well, and then stuffed in to 26 mm casing. The links were formed into sections and frozen.

[0733] To reheat, frozen sausage link were placed in a non-stick skillet with light oil spray and cooked over medium -low heat for 10-12 minutes, turning frequently to brown evenly, until internal temperature reached 165°F. They were covered with a lid to expedite heating. FIG. 44 is an example of the finished product.

Table 48. Euglena TVP Italian Dinner Sausage Formulation

Ingredient „ g Ί ^. % ^. I

. i

Example 21 - TVP Burger Patty

[0734] In this study, Water, Euglena TVP (Pea Protein, Euglena, Natural Flavor), Coconut Oil, Canola Oil, Seasoning (Onion Powder, Garlic Powder, Sugar, Natural Flavoring, Spice, Salt, Cooked Sugar, Brown Sugar, Sunflower Oil, Potato Maltodextrin, Silicon Dioxide, Com Maltodextrin), Methylcellulose, Citrus Fiber, Mung Bean Protein, Black Malt Extract, Konjac Gum, Natural Flavor, and Sea Salt were combined to create a TVP Burger Patty comprising of approximately 17% Euglena TVP.

[0735] Methods and Materials: In this study, the formulation used to create the TVP Burger Patty is listed in (Table 49), which includes approximately 17% of Euglena TVP. TVP was chopped while dry, to fine granule consistency. TVP and Citri-Fi TX10 were dry-blended, then boiling water was added, stirred well and let chill in cooler. TVP was combined at mixer. It was mixed on low 1 minute to combine. Citri-Fi 100FG and Dadex AR-HX (Antioxidant) were dissolved into liquid coconut oil and water was added. The mixture was emulsified with stick blender. It was chilled 24 hours. The mixture was combined with TVP mixture in stand mixer with paddle attachment. It was mixed for 2 minutes, until thoroughly dispersed. It was addedto the mixture in stand mixer, mixed on low, and canola oil was slowly added. Once evenly dispersed, the methylcellulose was added. It was mixed on low for approximately 3-5 minutes until mass was sticky and cohesive. The sides of the bowl were scraped down while mixing. The mixture was chilled, and then formed into 56 g patties. They were stored frozen.

[0736] To reheat, frozen patties were placed in non-stick skillet with light oil spray and cooked over medium-low heat for approximately 10-12 minutes, turning frequently to brown evenly, until internal temperature reached 165°F. They were covered with a lid to expedite heating. FIG. 45 is an example of the finished product. Table 49. Euglena TVP Burger Patty/Crumble Formulation

[0737] SUMMARY

[0738] The Euglena-based TVP is a versatile product and can be used in a variety of products mirroring the properties of a natural animal-based protein. In both looks, flavour and texture, the Euglena-based TVP can be used in application in which poultry, fish, beef and/or pork are typically used. The Euglena based TVP can also be used for any application where a vegetarian or vegan based meat analogue is used, for example traditional texturized vegetable protein. Market comparison has found that the Euglena-based TVP contains less and/or no allergens than most products currently on the market to date.

Example 22 - High moisture meat analogues (HMMA) using a basis of pea protein and different levels of wet Euglena biomass and Euglena protein rich flour

[0739] Introduction

[0740] High-moisture extrusion (HME) can create a product with fibrous texture like animal meat, known as a high-moisture meat analogue (HMMA), from plant protein raw materials. This process is a better method than conventional low-moisture extrusion for improving the texture of meat and seafood analogue. During the HME process, proteins undergo thermal and mechanical stresses by heating of the barrel and shearing of the screws. In consequence, protein structure is altered leading to the formation of soluble and/or insoluble aggregates. In the cooling die, the protein molecules are cross-linked, leading to the formation of a fibrous meat-like structure. Soy protein is the most studied plant protein in extrusion cooking, because of its functionality and nutritional properties. However, there are disadvantages of using soybeans, due to being a common food allergen and cultivation using genetically modified organisms (GMOs).

|0741] In our study, pea protein and Euglena protein were used instead. E. gracilis has been shown to produce protein containing all 20 proteinogenic amino acids. Moreover, animal and in vitro studies have shown an excellent digestibility of protein derived from E. gracilis biomass.

[0742] Objective

[0743] The objective of this study was to produce meat analogues with the combination of Euglena protein and pea protein by extrusion technology and to compare HMMA made with different pea protein isolates, and made with different inclusion levels of Euglena flour or thawed Euglena biomass.

[0744] Materials and Equipment

[0745] Materials: Euglena protein rich flour, wet biomass, masker, pea protein isolate (1), pea protein isolate (2), pea starch, pea fibre.

[0746] Materials used to generate the High Moisture Meat Analogue (HMMA) can be found in Table 50 which comprises: a starch, fiber, masker, Euglena source and a pea protein source. Materials were generated using an Clextral EV 32 twin screw extruder that was equipped with a specific screw profile. The HMMA setup included the full set of cooling dies and parameters can be found in Table 51.

Table 50. HMMA Formulations using 2 different sources of biomass (wet biomass, dry flour) and 2 different Pea Protein isolate sources.

Table 51. Extrusion parameters for making HMMA

[0747] Results: For the HMMA formulation 1, it was the most yellow ofthe 4 trials, with a very mild fishy lEuglena taste, slightly undercooked, and with a floury and glue like consistency. HMMA Formula 2 was lighter in colour compared to formulation 1, blander, but with higher elasticity and tenderness. Formulation 3 was darker than the other 2 formulations, mild fish taste with a bit of bitterness and metallic taste. This was more elastic than the formulations 1 & 2 Lastly, formulation 4 was the lightest in colour, cleaner than 3 with no bitterness and the most elastic (FIG. 46).

|0748] Conclusion: Overall, compared to the last HMMA Trial in May, the sensory properties of this HMMA were improved. For texture, it had longer fibers that were more uniform in shape and size, mostly long and narrow strips, as well as consistent looking fiber appearance. For flavor, it had milder marine flavor or no marine flavor detected. Also, it was lighter in color.

[0749] HMMA Formula 1 (made with pea protein isolate with wet biomass, without Euglena flour) had the most positive feedback because of the desired flavor and texture. HMMA Formula 3 and 4 (made with pea protein isolate, without wet biomass, with Euglena flour) had a very rubbery texture. All formulas had mild or no fishy flavor.

Example 23 - Additional example of texturized protein made with Euglena flour

[0750] Introduction: Extrusion is a common industrial process to produce a texturized protein as the final product. This process changes the characteristics of the starting ingredients causing a change in density, shape, appearance, taste and mouth feel.

[0751] To generate this extruded product, a mixture such as, protein and/or fiber, and/or fat, and/or other choice additives was mixed with water or treated with a preconditioner. The mixture was then extruded under heat and pressure at a specific speed head to generate a texturized protein product. Texturized products can vary in size and shape from small to larger sized chunks by using different dies and/or cutter speeds.

[0752] In this example, a Euglena based extruded product using Euglena Flour produced a texturized protein, which is referred to as a Texturized Vegetable Protein (TVP) or a Texturized Euglena Protein (TEP). In addition, the testing of an Urschel comitrol processor and die face cutter to generate various sizes of texturized proteins was also investigated. Euglena gracilis has been shown to produce protein containing all 20 proteinogenic amino acids. Moreover, animal and in vitro studies have shown an excellent digestibility of protein derived from Euglena gracilis biomass.

[0753] Objective: The objective of this study was to produce extruded texturized protein that can be used in the formation of meat analogues by using Euglena flour and pea protein. As well, the use of an Urschel comitrol processor on TVP or die face cutters speed was investigated. [0754] Materials and Equipment: Materials: Euglena protein rich flour, masker, pea protein concentrate, pea protein isolate (1).

[0755] Materials used to generate the texturized protein can be found in Table 52 which comprises: a protein concentrate source, a masker, a Euglena source, and a protein isolate source. In future, Euglena Protein Concentrate and Protein Isolate can be used to replace the ones tested in this formulation.

[0756] Materials were generated using an Clextral EV 32 twin screw extruder that was equipped with a specific screw profile for TVP. No preconditioner was used for the TVP. In TVP formula 1, no preconditioner was used but it did show that more time was needed to have the flour absorb the moisture. In TVP formula 2, the premix was tempered with water (i.e. 30 kg premix to 5.29 kg of water) and allowed to absorb the moisture more before feeding. A cutter was added to the end of the die, and an Urschel comitrol processor was used to test the shredding of the TVP. A hot air drier was used to dry the TVP products in trays for 40 minutes at 80 °C after forming. The TVP setup can be found in Table 53.

Table 52. Euglena texturized protein (TVP) formulations using Pea Protein Concentrate, masker, Euglena Flour, and Pea Protein Isolate.

Table 53. Extrusion parameters for making Euglena TVP with Euglena flour.

[0757] Results: Two different formulations were evaluated: TVP Formula 1 (FIG. 47) had the higher Euglena flour inclusion at 13% while TVP formula 2 (FIG. 48) was 10%. With respect to sensory observations, it was noted that the colour of TVP Formula 1 was lighter than previously completed trials; taste wise, there was a slight fishy taste but overall, the flavour was found to be acceptable. TVP Formula 2 was observed have an even lighter colouring than that of TVP Formula 1, with the taste of TVP Formula 2 being better than TVP Formula 1, with no fishy flavour noted, and it was also found to be cleaner than TVP Formula 1, pre hydrated. While there were differences noted in colour and flavour, the texture of both formulas was found to be chewy, springy and spongy.

[0758] Two different speeds were used to generate different sized TVP products. A slower speed was used to make the larger chunk product, while a faster speed was used to generate the smaller sized die cut products. Finally, the Urschel processor was used to shred the texturized protein into even smaller pieces and a large number of fines was produced. Both TVP Formula 1 and 2 were successful in producing texturized protein samples that can be used in meat analogue applications.

[0759] Conclusion: Texturized protein was successfully created with Euglena flour. In terms of ingredients that can be used to generate texturized protein, on a dry basis Euglena flour can be used, or can be replaced with wet biomass (either fresh from production or from frozen and thawed). Protein concentrate and protein isolate from Euglena can also be used in the future as replacements for pea protein and/or as a replacement for the Euglena flour. In addition, Beta Glucan Isolate (BGI) can be added in order to increase fiber and change the consistency of the extruded material. Wet or dry ingredients can be used. Other sources of protein concentrate and/or isolate can be used from other plant based sources such as: soy, chickpea, lentils, almond, legume, quinoa, millet, sorghum, teff, or any other reasonable flour, protein concentrate or protein isolate source.

[0760] Die cut heads, cutter speeds and processors such as the Urschel can be used to change the size and shape of the extruded Euglena protein. These Euglena TVP materials can be used to replace chicken, beef, pork, seafood, and even other plant based texturized proteins in various meat applications.

[0761] Experiment 1 : Larger Scale batch of extruded Euglena protein

[0762] Objective: To produce a larger batch scale of extruded protein made with Euglena flour and to test different extruded shapes.

[0763] Materials and Methods:

[0764] Materials: Euglena protein rich flour, masker, pea protein concentrate, pea protein isolate (1).

[0765] Materials used to generate the texturized protein can be found in Table 54 which comprises: a protein concentrate source, a masker, a Euglena source, and a protein isolate source. In future, Euglena Protein Concentrate and Protein Isolate can be used to replace the ones tested in this formulation.

[0766] Prior the extrusion, the flour mixture was mixed in a Ribbon blender. A Clextral EV 44 twin screw extruder that was equipped with a preconditioner was used to make the extruded product. It was dried in a continuous 5 layer steam drier and the TVP was reduced in size by the use of a roller mill. Flour bulk density was 384 g/L and moisture content was 4.69%. The TVP setup can be found in Table 55.

Table 54. Euglena texturized protein (TVP) formulation 3

Table 55. Extrusion parameters for making Euglena TVP with Euglena flour.

[0767| Results and Discussion:

[0768] A total of 290 kg (150, 120, 120 kg) was produced. Roll milling was used to produce smaller pieces with a bulk density of 98.4 g/L.

[0769] The disclosures of each and every patent, patent application, publication, and accession number cited herein are hereby incorporated herein by reference in their entirety.

[0770] Preferences and options for a given aspect, feature, embodiment, or parameter of the invention should, unless the context indicates otherwise, be regarded as having been disclosed in combination with any and all preferences and options for all other aspects, features, embodiments, and parameters of the invention.

[0771] While present disclosure has been disclosed with reference to various embodiments, it is apparent that other embodiments and variations of these may be devised by others skilled in the art without departing from the true spirit and scope of the disclosure. The appended claims are intended to be construed to include all such embodiments and equivalent variations.