TECHNICAL FIELD

[0001 ] The present invention generally relates to plant-based nutritional compositions comprising cereal starch hydrolyzed by enzymes produced by a koji mold or an enzyme preparation derived from a koji mold. In particular, the invention relates to plant-based nutritional compositions comparable in protein quality to dairy analogues, preferably wherein said nutritional compositions comprise little or no added sugar and are non-dairy.

BACKGROUND

[0002] Dairy foods are an important nutrient dense constituent of a healthy diet due to their capacity to provide essential vitamins, minerals, macronutrients and micronutrients important for growth, development and tissue maintenance. Yet, over time, there are segments of the human population that are increasingly avoiding consuming dairy foods. There are several reasons for this. Firstly, given that lactose is a primary sugar present in animal milk, those who are lactose intolerant can suffer abdominal pain, bloating, diarrhea, gas, and nausea due to the ingestion of animal milk-based dairy products. Lactose intolerance arises from the inability to digest lactose due to a relative lack of the lactase enzyme, which is the enzyme normally present in the human gut that is responsible for hydrolysis of lactose to glucose and galactose. According to the US National Institutes of Health (NIH), lactose intolerance affects more than half of the Americans adults, many of whom are people with Asian or African heritage (see https://Qhr.nlm.nih.qov/condition/lactose-intolerance).

[0003] Secondly, dairy foods may also contain significant amounts of cholesterol and saturated fat. Accordingly, such foods are not recommended for individuals suffering from heart diseases.

[0004] Thirdly, dairy foods may be supplemented with high amounts of sucrose and are therefore not considered suitable for those who are obese or who have or are susceptible to diabetes. [0005] The above reasons point to the evolution of plant-based foods as being a significant alternative to dairy foods. In particular, plant-derived milk analogues and fermented non-dairy analogues (yogurt analogues, non-dairy beverages and cheese analogues) are becoming increasingly prominent on supermarket shelves. [0006] This being said, plant-based proteins are often not as good in quality as dairy proteins. The protein digestibility-corrected amino acid score (PDCAAS) is a method of evaluating the quality of a protein based on both the amino acid requirements of humans and their ability to digest it. PDCAAS compares the amount of the essential amino acids in a food to a reference (scoring) pattern based on the essential amino acid requirements of a preschool-age child to determine its most limiting amino acid (amino acid score). This approach is recommended by the Food and Drug Administration (FDA) and is described in the 1991 FAO/WHO Protein Quality Report. Using the PDCAAS method, the highest possible score is a 1.0 meaning, that after digestion, a protein having a PDCAAS of 1 .0 provides (per unit of protein) 100% or more of the indispensable amino acids required. As examples, whereas cow's milk and casein (milk protein) have a PDCAAS of 1.0, the PDCAAS of many plant-based proteins is often less than 1.0. [0007] It would therefore be desirable to provide a plant-based nutritional composition, which is free from any dairy ingredients, having a satisfactory texture and taste, an improved PDCAAS and little or no added sugar.

[0008] Discussion or mention of any piece of prior art in this specification is not to be taken as an admission that the prior art is part of the common general knowledge of the skilled addressee of the specification.

SUMMARY

[0009] This summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This summary is not intended to identify key features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter. [0010] In view of the above reasons for avoiding dairy foods, improved plantbased foods that have a protein quality, as measured by the PDCAAS method, that is equivalent to a dairy-food counterpart would be beneficial. Such plant-based foods should preferably have little or no added sugar.

[0011 ] According to one aspect, the invention provides a process for producing a nutritional composition comprising the steps of: providing a hydrolyzed cereal slurry comprising cereal starch hydrolyzed by enzymes produced by a koji mold or an enzyme preparation derived from a koji mold; and fermenting the hydrolyzed cereal slurry with at least one lactic acid-producing bacteria, thereby producing a nutritional composition.

[0012] In an embodiment, the koji mold comprises at least one microorganism of the genus Aspergillus, preferably at least one Aspergillus selected from the group consisting of A. oryzae, A. kawachii, A. awamori, and A. luchuensis.

[0013] In another embodiment, the process comprises combining a cereal slurry with the koji mold producing said enzymes for producing said hydrolyzed cereal slurry. Alternatively, the process may comprise combing the cereal slurry with the enzyme preparation derived from the koji mold for producing said hydrolyzed cereal slurry. The invention has the advantage that rice koji may be directly used and thereby bypassing an extra step of enzyme production as rice koji already contained enzymes. Furthermore, according to the present invention rice koji may advantageously be used with more broad covering enzymes and not only with a single enzyme.

[0014] In another embodiment, the at least one lactic acid-producing bacteria is selected from the group consisting of: Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, Streptococcus, Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus, and Weissella, preferably selected from the group consisting of Lactobacillus, Lactococcus, Streptococcus, and Bifidobacterium, further preferably selected from the group Lactobacillus, Streptococcus and Lactococcus, most preferably Streptococcus and Lactobacillus, and optionally wherein fermenting hydrolyzed cereal slurry with said at least one lactic acid-producing bacteria is performed until a pH between 4.0 and 5.0, preferably between 4.0 and 4.5, is achieved.

[0015] Other embodiments of nutritional compositions according to the present invention will be evident from the following detailed description.

DESCRIPTION OF FIGURES

[0016] Figure 1 is a flow diagram of an exemplary process of producing a nutritional composition according to the present invention.

[0017] Figure 2 shows acidification curves for various plant-based nutritional compositions during fermentation with lactic-acid producing bacteria over a 7hr period. The top line is the acidification curve of a composition comprising coconut cream and pea protein without added sugar. The middle line is the acidification curve of a composition comprising coconut cream, pea protein with added sugar. The bottom line is the acidification curve of a composition comprising coconut cream, pea protein and oat slurry without added sugar.

[0018] Figure 3 shows the results of test to determine the rheological properties, especially the firmness, of set (clear bars) and stirred (shaded bars) oat-based non-dairy yogurt analogues.

DETAILED DESCRIPTION

Definitions and statements regarding interpretation

[0019] The terms “a,” “an,” “the” and similar referents used in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0020] All methods and processes described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. [0021 ] The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0022] Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. For example, if a range is from about 1 to about 50, it is deemed to include, for example, 1 , 7, 34, 46.1 , 23.7, or any other value or range within the range.

[0023] Unless noted otherwise, all percentages in the specification refer to weight percent, where applicable.

[0024] The terms "comprise", "comprises", "comprised" or "comprising", "including" or "having" and the like in the present specification and claims are used in an inclusive sense, that is to specify the presence of the stated features but not preclude the presence of additional or further features.

[0025] Specific embodiments disclosed herein may be further limited in the claims using consisting of or consisting essentially of language. When used in the claims, whether as filed or added per amendment, the transition term “consisting of” excludes any element, step, or ingredient not specified in the claims. The transition term “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). Embodiments of the invention so claimed are inherently or expressly described and enabled herein.

[0026] Unless defined otherwise, all technical and scientific terms have and should be given the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Plant based-nutritional compositions [0027] In one aspect, the present invention provides processes for producing plant-based nutritional compositions, preferably non-dairy plant-based compositions. In embodiments, the nutritional compositions of the present invention are derived from at least one cereal selected from the group consisting of: oats, barley, rice, wheat, maize, rye, sorghum, triticale, sesame, quinoa, buckwheat, spelt, amaranth, and pearl millet, most preferably oats.

[0028] In embodiments, the processes of the present invention comprise milling at least one cereal to produce a milled cereal. Milling may be any conventional milling procedure, including dry or wet milling. Dry milling may produce a flour, which is mixed with at least one liquid producing an aqueous suspension. In other embodiments, the at least one cereal may be wet milled with at least one liquid to produce a cereal slurry. Therefore, in the present context, a cereal slurry is an aqueous cereal suspension produced by mixing at least one dry milled cereal with at least one liquid, or by wet milling at least one cereal with at least one liquid. The at least one liquid for producing the cereal slurry is preferably water. However, liquids comprised of oils or fats (such as plant-based edible oils and fats, or oil/fat comprising emulsions) may also be suitable. Equally, the processes of the present invention also encompass producing low or no fat nutritional compositions. The liquid and the solids in the cereal slurry are not separated.

[0029] In preparing the slurry, cereal starches are released. In embodiments, the processes of the present invention comprise treating the slurry with an enzyme preparation for enzymatic hydrolysis of said cereal starches. The enzymatic preparation preferably comprises at least one hydrolase enzyme having the ability to hydrolyze a-glycosidic bonds. The hydrolases may be selected from the group consisting of [3-amylase, a-amylase, amyloglucosidase (glucoamylase), pullulanase and mixtures thereof. Preferably, when there are two or more enzymes combined in the enzyme preparation, the enzymes are introduced simultaneously into the cereal slurry. In addition to the above-identified hydrolases, other enzymes may optionally be present. In embodiments, the enzyme preparation may further comprise an isomerase, such as glucose isomerase, a peptidase such as a carboxypeptidase, and/or a tyrosinase.

[0030] Preferably, the enzymatic preparation is prepared with enzymes isolated from koji mold. Koji mold comprises spores of at least one microorganism of the genus Aspergillus, preferably at least one Aspergillus selected from the group consisting of A. oryzae, A. kawachii, A. awamoh, and A. luchuensis, most preferably A. oryzae. Enzymes may be obtained by known methods, including by culturing said at least one Aspergillus and isolating secreted enzymes from the culture supernatant by filtration. Alternatively, genes encoding Aspergillus-de ved enzymes may be cloned and expressed using an in vitro expression system (eg bacteria or yeast), facilitating enzyme isolation.

[0031 ] Cereal starch hydrolysis using the enzymatic preparation is preferably performed under optimal conditions permitting hydrolysis. In embodiments, the temperature for cereal starch hydrolysis using the enzymatic preparation is at least 40°C, preferably between 50°C to 80°C, most preferably between 55°C to 75°C. In embodiments, the pH for cereal starch hydrolysis using the enzymatic preparation is at least 4.0, preferably between 4.0 to 6.0, most preferably between 5.0 to 5.8. In embodiments, the time for cereal starch hydrolysis using the enzymatic preparation is at least 4 hrs, preferably between 4 hrs to 8 hrs, most preferably between 5 hrs to 7 hrs.

[0032] Alternatively, a more 'natural' approach to cereal starch hydrolysis may be adopted. Specifically, the slurry comprising cereal starch may be hydrolyzed in the presence of at least one hydrolase-producing microorganism. In some embodiments, the processes of the present invention comprise hydrolysis of the slurry comprising cereal starch by koji mold. As noted above, koji mold comprises spores of at least one microorganism of the genus Aspergillus, preferably at least one Aspergillus selected from the group consisting of A. oryzae, A. kawachii, A. awamoh, and A. luchuensis, most preferably A. oryzae. In embodiments, the koji mold is dry rice koji mold. Preferably, dry rice koji mold is prepared using dry- milled rice. In embodiments, the dry rice koji is prepared by soaking, boiling and cooling said milled rice, and subsequently inoculating and incubating said rice with koji mold. When sufficient growth is achieved, the mixture is then dried, providing dry rice koji mold. In embodiments, dry rice koji mold comprises at least 50,000 cfu/g of mold, preferably at least 100,000 cfu/g of mold, most preferably 150,000 cfu/g of mold.

[0033] In embodiments, said at least one hydrolase-producing microorganism also produces proteolytic enzymes which contribute to digestion of cereal proteins in said cereal slurry. Proteolytic enzymes preferably include endo- and/or exopeptidases, most preferably carboxypeptidases.

[0034] In embodiments, cereal starch hydrolysis is performed at temperatures permitting koji fermentation - that is, temperatures that facilitate growth and active fermentation by koji mold, said mold comprising at least one microorganism of the genus Aspergillus. Preferably, during koji fermentation, said at least one microorganism releases enzymes that hydrolyze said cereal starch. In embodiments, the temperature for cereal starch hydrolysis using koji fermentation is at least 25°C, preferably between 30°C to 45°C, most preferably between 30°C to 40°C. In embodiments, the pH for cereal starch hydrolysis using koji fermentation is at least 4.0, preferably between 4.0 to 6.0, most preferably between 5.0 to 5.8. In embodiments, the time for cereal starch hydrolysis using koji fermentation is at least 4 hrs, preferably between 4 hrs to 8hrs, most preferably between 5 hrs to 7 hrs.

[0035] In alternative embodiments, cereal starch hydrolysis is performed at temperatures that do not allow koji mold microorganism growth and active fermentation. Specifically, microorganisms that facilitate koji fermentation are typically unable to grow at temperatures greater than 44°C. Nevertheless, cereal starch hydrolysis may still occur at temperatures greater than 44°C. In embodiments, starch in a cereal slurry inoculated with koji mold may be hydrolyzed by enzymes contained in and released by said mold. That is, the temperature may not allow koji mold growth and active fermentation, but will allow hydrolysis by way of enzymes that are contained in and released by the koji mold microorganisms. In embodiments, the temperature for cereal starch hydrolysis using koji mold under conditions that do not permit microorganism growth and active fermentation is at least 45°C, preferably between 50°C to 80°C, most preferably between 55°C to 75°C. In embodiments, the pH for cereal starch hydrolysis using koji mold under conditions that do not permit microorganism growth and active fermentation is at least 4.0, preferably between 4.0 to 6.0, most preferably between 5.0 to 5.8. In embodiments, the time for cereal starch hydrolysis using koji mold under conditions that do not permit microorganism growth and active fermentation is at least 4 hrs, preferably between 4 hrs to 8 hrs, most preferably between 5 hrs to 7 hrs.

[0036] In further embodiments, the processes of producing plant-based nutritional compositions of the present invention may incorporate the step of hydrolyzing said cereal starch (by addition of an enzymatic preparation and/or by 'natural' microorganism fermentation). In alternative embodiments, the processes comprise provision of a slurry in which the cereal starch is already hydrolyzed (ie pre-hydrolyzed).

[0037] The cereal (or combinations of cereals) may beneficially comprise nonstarch polysaccharides. For example, 58% of the total non-starch polysaccharides in oats are soluble [3-glucans. [3-glucans are known to have beneficial impact on insulin resistance, hypertension, and obesity. [3-glucans are also reported to lower blood cholesterol and thus is commonly used in functional food products.

[0038] Apart from fibers, cereals such as oats also contain vitamins such as thiamin, riboflavin, niacin, vitamin B6, folate, pantothenic acid and vitamin E. Cereals may also contain minerals depending on the soil content in which they are grown, said minerals including calcium, copper, iron, magnesium, manganese, phosphorus, selenium and zinc.

[0039] Optionally, the processes of the present invention comprise a step where the hydrolyzed cereal slurry is subsequently pasteurized so as to significantly reduce and/or kill microorganisms that could cause disease, spoilage, or undesired fermentation. In embodiments, pasteurization is achieved by heating the hydrolyzed cereal slurry to at least 80°C, preferably at least 90°C, most preferably at least 95°C. The time to achieve pasteurization is at least 5min, preferably at least 10min, most preferably at least 15min. [0040] Optionally, the non-hydrolyzed or hydrolyzed cereal slurry is homogenized before pasteurization. In embodiments, homogenization occurs at a pressure from 50 bar to 700 bar, preferably from 100 bar to 300 bar, further preferably from 150 to 300 bar, most preferably at 200 bar. Without wishing to be bound by theory, it is believed that homogenization functionalizes the plant proteins. For example, homogenization may produce a self-supporting gel resulting from the coagulation of plant proteins. A self-supporting gel forms a satisfactory texture mimicking the texture of standard dairy yogurts.

[0041 ] The hydrolyzed cereal slurry may be utilized as a base for preparing a variety of plant-based nutritional compositions of different forms. In embodiments, the processes of the present invention comprise preparing plant-based nutritional compositions, such as non-dairy yogurt analogues and desserts (including frozen yogurts and desserts), and non-dairy beverages. Non-dairy or dairy-free products do not contain milk or milk products. Further, compositions according to the present invention may be of varied consistency. That is, compositions may be in a form that is spoonable (eg a set or stirred yogurt), squeezable, pourable, smearable/spreadable, cuttable, or otherwise edible.

[0042] In embodiments, the method of preparing the plant-based nutritional compositions of the present invention comprise fermenting the hydrolyzed cereal slurry comprising hydrolyzed cereal starch with at least one lactic acid-producing bacteria (LAB). Preferably, the at least one lactic acid-producing bacteria is selected from the group consisting of: Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, Streptococcus, Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus, and Weissella, preferably selected from the group consisting of Lactobacillus, Lactococcus, Streptococcus, and Bifidobacterium, and most preferably selected from the group Lactobacillus and Lactococcus. In embodiments, the fermentation of the hydrolyzed cereal slurry with at least one LAB is performed until reaching pH of at least 4.0, preferably between 4.0 and 5.0. In yet further embodiments, the fermentation of the hydrolyzed cereal slurry with at least one LAB is performed at a temperature lower than 50°C, most preferably at 40°C. [0043] In embodiments, the processes of preparing the plant-based nutritional compositions of the present invention further comprises fermenting the LAB- fermented slurry in the presence of: at least one yeast, preferably selected from the group consisting of: Zygosaccharomyces, Candida, Kloeckera/Hanseniaspora, Torulaspora, Pichia, Brettanomyces/Dekkera, Saccharomyces, Lachancea, Saccharomycoides, Schizosaccharomyces, and Kluyveromyces, most preferably Saccharomyces and Kluyveromyces, and/or at least one acetic acid-producing bacteria, preferably selected from the group consisting of Acetobacter and Gluconacetobacter. As such, the processes of the present invention encompass the preparation of plant-based fermented products, such as kefir and kombucha, respectively. These LAB and yeast fermented products are preferably non-dairy. [0044] In some embodiments, an advantage of utilizing a hydrolyzed cereal slurry for subsequent fermentation is that the hydrolyzed starch is used by the fermenting microorganisms as a sugar source. That is, the processes of the present invention advantageously provide the possibility of preparing nutritional compositions by limiting or avoiding additional sugar content, therefore reducing calories, while enabling fermentation. Preferably, in the processes of the present invention, no greater than 10% w/w additional sugar is added, further preferably wherein no greater than 5% w/w additional sugar is added. Most preferably, the processes of the present invention exclude the addition of additional sugar. In embodiments, fermentable sugars present in the hydrolyzed cereal slurry comprise one or more of glucose, fructose, sucrose, maltose, and maltotriose, preferably glucose and maltose, most preferably glucose.

[0045] In embodiments, the processes of the present invention may comprise preparing a plant-based nutritional composition that predominantly comprises a hydrolyzed cereal slurry, excluding other plant-based extracts. Alternatively, in other embodiments, the hydrolyzed cereal slurry may be mixed with at least one plant-based protein and/or at least one plant-based liquid extract.

[0046] In embodiments, the at least one plant-based protein is at least one legume protein. A legume is a plant in the family Fabaceae (or Leguminosae). When used as dry grain, the seed of a legume is called a pulse. Preferably, the at least one legume protein is obtainable from the group consisting of beans, garbanzo, faba beans, pea, lentil, chana dal and soybean, most preferably wherein the at least one additional protein is pea protein.

[0047] In embodiments, the at least one liquid is plant-based, preferably at least one plant-based milk, cream, water, oil and/or emulsion (eg an oil-in-water emulsion), or mixtures thereof, further preferably a coconut cream, coconut milk, or coconut water, most preferably a coconut cream.

[0048] In yet other embodiments, the hydrolyzed slurry may be mixed with at least one plant-based protein and at least one plant-based liquid extract.

[0049] In some embodiments, the mixing of the slurry with other plant-based extracts is preferably a step after the hydrolysis of cereal starch, but before the subsequent fermentation step (ie wherein subsequent fermentation is in the presence of at least one LAB, optionally comprising at least one yeast and/or at least one acetic acid-producing bacteria), and preferably before homogenization and, pasteurization, if such steps are employed.

[0050] In some embodiments, the processes of the present invention comprise producing a nutritional composition that comprises between 10% (w/w) to 50% (w/w) fermented slurry (ie LAB fermented, optionally fermented by yeast and/or acetic acid-producing bacteria), preferably between 15% (w/w) to 35% (w/w), most preferably between 20% (w/w) to 30% (w/w) of said fermented slurry. [0051 ] In some embodiments, the processes of the present invention comprise producing a nutritional composition that comprises between 1 % (w/w) to 10% (w/w) of the at least one plant-based protein, preferably between 3% (w/w) to 7% (w/w), most preferably between 4% (w/w) to 6% (w/w).

[0052] In some embodiments, the processes of the present invention comprise producing a nutritional composition that comprises between 5% (w/w) to 35% (w/w) of the plant-based liquid extract, preferably between 10% (w/w) to 30% (w/w), most preferably between 12% (w/w) to 20% (w/w).

[0053] The protein digestibility-corrected amino acid score (PDCAAS) is a method of evaluating the quality of a protein based on both the amino acid requirements of humans and their ability to digest it. Specifically, the method is predicated on the understanding that protein quality can be assessed by expressing the content of the first limiting essential amino acid of the test protein as a percentage of the content of the same amino acid in a reference pattern of essential amino acids. This reference pattern is based on the essential amino acid requirements of the preschool-age child. The formula for calculating PDCAAS percentage is as follows:

PDCAAS = (mg of limiting amino acid in 1 g of test protein I mg of same amino acid in 1 g of reference protein) x fecal true digestibility percentage

[0054] Oat-based nutritional compositions typically have a PDCAAS of less than 0.6. Accordingly, processes of the present invention advantageously provide nutritional compositions comprising a PDCAAS score of 0.6 or higher and comparable to dairy analogues. In some embodiments, the PDCAAS of the nutritional composition produced by the processes of the present invention is at least 0.6, preferably between 0.6 to 1 , most preferably between 0.7 to 0.99. In other embodiments, where the processes of the present invention produce a nutritional composition comprising a mixture of an oat slurry and pea protein, the PDCAAS of said composition is at least 0.90, preferably at least 0.95.

[0055] In some embodiments, nutritional compositions of the present invention which comprise oat slurry, but exclude added protein and sugar, comprise the following contents (%w/w):

[0056] In some embodiments, in the processes of the present invention, before the hydrolyzed cereal slurry is subjected to the subsequent LAB fermentation (optionally in the presence of yeast and/or acetic acid bacteria), said slurry comprises: at least 5% (w/w) glucose, preferably between 5% (w/w) to 20% (w/w), most preferably between 8% (w/w) to 15% (w/w); and at least 2% (w/w) maltose, preferably between 2% (w/w) to 6% (w/w), most preferably between 3% (w/w) to 5% (w/w).

[0057] As mentioned above, the processes of the present invention may produce any number of plant-based nutritional compositions (eg spoonable, squeezable, pourable, smearable/spreadable, cuttable and the like) comparable to their dairy counterparts in protein quality as well as texture and taste. Preferably, where the processes are used to produce a yogurt, the firmness of the yogurt when set is at least 0.4 N, preferably at least 0.5 N, when tested by back-extrusion using a 30 mm cylindrical flat probe penetrated into a sample of said set yogurt at a crosshead speed of 0.5 mm/s to a depth of 30 mm, whereby the mean force between 15 mm and 25 mm is obtained. Further preferably, the firmness of the yogurt when stirred is at least 0.1 N, preferably at least 0.2 N, when tested by back- extrusion using a 50 mm cylindrical flat probe penetrated into a sample of said stirred yogurt at a crosshead speed of 0.5 mm/s to a depth of 30 mm, whereby the mean force between 15 mm and 25 mm is obtained. Preferably, firmness is determined using a pseudo-compression (“back-extrusion”) test using a TAX-T2 Texture Analyzer (TA Instruments, Stable Micro Systems, UK).

[0058] In another aspect, the present invention provides nutritional compositions produced by the processes described herein (ie product-by- process). [0059] In yet another aspect, the present invention provides nutritional compositions comprising a hydrolyzed cereal slurry hydrolyzed by koji mold or enzymes extracted from koji mold, preferably a hydrolyzed oat slurry, wherein said slurry is fermented by at least one LAB, thereby producing the nutritional composition. In embodiments, LAB fermentation is performed until reaching a pH between 4.0 and 5.0, preferably between 4.0 and 4.5.

[0060] In embodiments, the present invention provides nutritional compositions comprising a mixture of: a hydrolyzed cereal slurry hydrolyzed by koji mold or enzymes derived from koji mold, preferably a hydrolyzed oat slurry; at least one plant-based protein, preferably pea protein, and optionally a plant-based milk or cream extract, preferably a coconut cream; wherein the mixture is fermented by at least one LAB, thereby producing the nutritional composition. In embodiments, LAB fermentation is performed until reaching a pH between 4.0 and 5.0, preferably between 4.0 and 4.5.

[0061 ] Optionally, fermentation by at least one LAB may further comprise at least one yeast and/or at least one acetic acid bacteria. Preferably, the at least one lactic acid-producing bacteria is selected from the group consisting of: Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, Streptococcus, Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus, and Weissella, preferably selected from the group consisting of Lactobacillus, Lactococcus, Streptococcus, and Bifidobacterium, and most preferably selected from the group Lactobacillus and Lactococcus. Said at least one yeast is preferably selected from the group consisting of: Zygosaccharomyces, Candida, Kloeckera/Hanseniaspora, Torulaspora, Pichia, Brettanomyces/Dekkera, Saccharomyces, Lachancea, Saccharomycoides, Schizosaccharomyces, and Kluyveromyces, most preferably Saccharomyces and Kluyveromyces. Said at least one acetic acid-producing bacteria is preferably selected from the group consisting of Acetobacter and Gluconacetobacter.

[0062] In embodiments, the nutritional compositions of the present invention comprise: between 10% (w/w) to 50% (w/w) hydrolyzed slurry, preferably between 15% (w/w) to 35% (w/w), most preferably between 20% (w/w) to 30% (w/w); between 1 % (w/w) to 10% (w/w) of the at least one plant-based protein, preferably between 3% (w/w) to 7% (w/w), most preferably between 4% (w/w) to 6% (w/w); and optionally between 5% (w/w) to 35% (w/w) of the plant-based liquid extract, preferably between 10% (w/w) to 30% (w/w), most preferably between 12% (w/w) to 20% (w/w).

[0063] In embodiments, the nutritional compositions of the present invention may further comprise other additives such as fruit preparations, flavors, colors, prebiotics, probiotics, vitamins, minerals, solid or soft inclusions, sauces, herbs and/or sources of oils and/or fats.

[0064] In embodiments, the nutritional compositions of the present invention consist of hydrolyzed oat slurry, water and at least one LAB. In other embodiments, the nutritional compositions of the present invention consist of hydrolyzed oat slurry, pea protein, water, at least one LAB, and optionally a coconut cream.

[0065] In embodiments, the nutritional compositions of the present invention are in spoonable, squeezable, pourable, smearable/spreadable, or cuttable form, including yogurts, desserts and the like, or beverages, preferably non-dairy products, most preferably a non-dairy yogurt.

[0066] Those skilled in the art will understand that they can freely combine all features of the present invention disclosed herein. Further, features described for different embodiments of the present invention may be combined.

[0067] Furthermore, where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred in this specification.

[0068] Further examples of the invention are described below. However, it should be noted that the invention should not be limited to these examples, and that the invention is susceptible to variations, modifications and/or additions other than those specifically described, and it is to be understood that the invention includes all such variations, modifications and/or additions which fall within the scope of the claims.

Examples

Example 1 - Preparation of a hydrolyzed oat slurry by koi i mold

[0069] Reference is made to Figure 1 which shows a process of producing a fermented oat-based yogurt. Oat flour (SWEOAT™ P14, Swedish Oat Fiber AB, Sweden) at 4.47 kg (26.3% w/w) is mixed with 10.73 kg boiling water (10.5% w/w) for 10-15 min until a homogeneous slurry is produced, then cooled to 60°C. Dry rice koji mold at 1.76 kg (63.1 % w/w) (Bio'c CO. LTD, Japan) is then mixed with the oat slurry until completely suspended. This mixture is incubated for approximately 6hrs at 60°C with intermittent mixing. An oat slurry comprising hydrolyzed starch is produced. This hydrolyzed slurry is used as a base for other nutritional compositions (see further below).

[0070] Tables 1 and 2 show the sugar content of respective oat slurry samples by ion chromatography according to the method of Ellingson, D., Anderson, P., Berg, D., “Analytical Method for Sugar Profile in Pet Food and Animal Feeds by High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection”, Journal of AOAC INTERNATIONAL 99 (2): 342-352 (2016).

Table 1 - Oat Slurry Sugar Analysis, Sample 1 - SweOat

Sugar profile by ion chromatography

[0071 ] After hydrolysis, the mixture is pasteurized at 95°C for 15m in. Once cooled to 40°C, the hydrolyzed oat slurry is mixed with 0.537 g (0.003% w/w) of a lactic acid-producing bacteria (LAB) DANISCO® VEGE 022 LYO 200 DCU

(consisting of Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus plantarum, Lactobacillus acidophilus and Bifidobacterium lactis). The amount of the fermenting microorganism may be varied depending on the types used (see Example 2 below, for example). [0072] The mixture is incubated at 40°C for 14hrs until a pH of less than 4.2 is achieved. The mixture is then chilled and stored at less than 8°C. Table 2 shows sugars and organic acids of the oat slurry after fermentation. It is clear that lactic acid is the dominant organic acid, and indicative of an equivalent fermentation to milk-based yogurts.

Table 2 - Oat Slurry Sugar Analysis, Sample 3

Sugar profile by ion chromatography

Organic Acids Example 2 - Preparation of an oat slurry mixture

[0073] The base hydrolyzed oat slurry (ie obtained from Example 1 after koji mold treatment and pasteurization) is mixed according to the recipes of Table 3 to produce fermented oat-based yogurt mixtures.

Table 3 - Oat-Based Yogurt Mixtures

[0074] The coconut cream is 24% fat aseptic coconut cream from First Grade international ltd. PURIS™ Pea 870 is yellow pea protein produced by Cargill, North America. YOFLEX® Mild 2.0, produced by CHR Hansen, contains Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophiles. Sugar is sucrose. [0075] The ingredients for each composition (coconut cream, water, pea protein, oat slurry) are mixed at 60°C and homogenized at approximately 200 bar until homogenous. The mixtures then undergo pasteurization at 92°C for 6 min, followed by cooling to 43°C. The mixtures are then inoculated with bacterial culture (YOFLEX® Mild 2.0) and incubated at 43°C until a pH of approximately 4.6 is obtained (around 6-7hrs). This is followed by smoothing at 25°C, then cooling and storage at 8°C.

[0076] Each composition comprises coconut cream and pea protein. Composition 1 comprises 5% w/w added sugar, whereas compositions 2 and 3 comprise no added sugar. In composition 3, oat slurry was added to 28.9% w/w to obtain a final 5% w/w total sugar. Advantageously, the sugar derived from koj i- mediated hydrolysis is the substrate for subsequent bacterial fermentation.

[0077] Figure 2 shows the acidification curves of the mixtures of Table 2 when inoculated with YOFLEX® Mild 2.0 and incubated at 43°C. This Figure shows that the inoculated bacteria grow and acidify each mixture, even without the addition of additional sugar (normally required as a food source by organisms responsible for fermentation). Notably, the addition of oat slurry at 28.9% w/w slightly acidified the mix before fermentation, and the target pH at the end of fermentation was advantageously obtained approximately 2 hrs earlier than with the other mixtures lacking oat slurry.

[0078] The taste and mouthfeel of the compositions are similar. All three are low in sweetness. While Composition 1 and Composition 2 are quite liquid, Composition 3 comprising the oat-slurry is thicker in mouthfeel. Further, the addition of oat slurry in Composition 3 provides nutty overtones that partially cover pea off-notes.

Example 3 - Rheological properties of oat slurry mixtures

[0079] The rheological properties of oat slurry mixtures of Example 2 is compared. For set yogurt gels, the mechanical properties are characterized based on a pseudo-compression (“back-extrusion”) test using a TAX-T2 Texture Analyzer (TA Instruments, Stable Micro Systems, UK). A 30mm diameter cylindrical flat probe penetrated into the sample at a crosshead speed of 0.5 mm/s and to a depth of 30mm. The mean force between 15 and 25mm obtained from this test is analyzed. For stirred yogurt gels, a similar protocol to the back-extrusion test used for set gels is employed, but with a 50 mm diameter cylindrical flat probe.

[0080] Figure 3 shows the results of the testing of set and stirred yogurt products. The absence of white sugar slightly decreased the firmness of set yogurts, due to lower total solids. Addition of oat slurry decreased the firmness of the stirred yogurt. Without being bound by theory, it is postulated that increased fiber and total solids partially inhibits gelation of pea proteins, disturbing the pea protein network formation.

Example 4 - Protein quality of oat and pea-based compositions

[0081 ] The amino acid profiles of PURIS™ Pea 870 and oat slurry as shown in Table 4 were used to estimate PDCAAS as shown in Table 5.

Table 4 - Amino Acid Profiles (g/100 g protein)

Method references:

R. Schuster, "Determination of Amino Acids in Biological, Pharmaceutical, Plant and Food Samples by Automated Precolumn Derivatization and HPLC", Journal of Chromatography, 1988, 431 , 271-284.

Henderson, J.W., Ricker, R.D. Bidlingmeyer, B.A., Woodward, C., "Rapid, Accurate, Sensitive, and Reproducible HPLC Analysis of Amino Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA columns and the Agilent 1100 HPLC," Agilent Publication, 2000. Barkholt and Jensen, "Amino Acid Analysis: Determination of Cysteine plus Half-Cystine in Proteins after Hydrochloric Acid Hydrolysis with a

Disulfide Compound as Additive", Analytical Biochemistry, 177, 318-322 (1989).

Henderson, J.W., Brooks, A., "Improved Amino Acid Methods using Agilent Zorbax Eclipse Plus C18 Columns for a Variety of Agilent LC Instrumentation and Separation Goals," Agilent Application Note 5990-4547 (2010).

Table 5 - Calculated PDCAAS

(a) Average of data provided by suppliers of pea proteins

(b) Data for Oat meal - "True Protein Digestibility Value of Common Foods." Federal

Register. 1993, Vol. 58, No. 3, 2193-2195.

[0082] Table 5 shows that the combination of pea protein and oat slurry greatly improves the PDCAAS, compared to either protein source separately. That is, the combination of pea protein and oat slurry provides a PDCAAS comparable to that of milk-protein.

Example 5 - Protein quality of other cereal and plant protein-based compositions [0083] PDCAAS values of other cereal protein combinations were determined as shown in Table 6. Faba Bean Protein 60 is produced by AGT Food and Ingredients (under the trade mark PulsePlus™ Protein).

[0084] Table 6 shows that other suitable cereal slurry-plant protein mixtures can achieve PDCAAS scores comparable to milk-protein based dairy products.

Table 6 - Calculated PDCAAS

Example 6 - Variation of total solids content

[0085] An experiment was performed to determine the extent to which varying the amount of rice koji mold would have on total solids (TS) content in an oat slurry. An oat slurry is produced according to Example 1 with oat flour and boiling water. Dry rice koji mold is then mixed with the oat slurry until completely suspended. This mixture is incubated for approximately 6hrs at 60°C with intermittent mixing. Table 7 shows a direct relationship between varying the amount of koji mold and total solids content (%w/w). The inventors have surprisingly found that reducing solids content can make subsequent processing of the oat slurry (eg during homogenization) easier.

Table 7 - Variation of oat slurry sugar content with varying amounts of rice koji

Conclusion [0086] Overall, processes according to the present invention, and compositions derived therefrom, are advantageous for at least the following reasons:

• It has been found that use of a microorganism (preferably a koji mold) or an enzyme preparation (preferably enzymes derived from a koji mold) to prepare a hydrolyzed cereal slurry enables the preparation of a nutritional composition in which hydrolyzed cereal starch is used as a sugar source for subsequent lactic acid bacteria, yeast and/or acetic acid producing bacteria fermentation. This minimizes or avoids the requirement to add sugar to support said bacterial/yeast fermentation. The use of whole microorganisms, as opposed to enzymic preparations, is preferred as a more natural approach for preparing a hydrolyzed cereal starch.

• It has also been found that processes of the present invention enable the preparation of nutritional compositions having a protein digestibility- corrected amino acid score (PDCAAS) equivalent to dairy analogues. This means that the protein quality, and therefore nutritional value, of compositions produced by processes according to the present invention is of equal or near-equal quality to a dairy analogue.

• Furthermore, it has been found that the processes of the present invention enable the preparation of nutritional compositions having a taste and/or texture equivalent to dairy analogues.

[0087] While illustrative embodiments have been illustrated and described, including the best mode known to the inventors for carrying out the invention, those skilled in the art will recognize that the invention may be practiced with variations on the disclosed structures, materials, compositions and methods, and such variations are regarded as within the ambit of the disclosure.