BACKGROUND

[0001] Plant-based protein products or meat substitutes have gained increasing market share as alternatives for meat not only as healthy meat alternatives but also for the reasons of them being made from more environmentally sustainable sources. Successful products on the market include, for example, IMPOSSIBLE FOODS™ and BEYOND MEAT™.

[0002] However, despite significant amount of innovation in the field of producing plant-based protein products or meat substitutes, there is still need to improve these products in regard to, for example, reducing additives that are used to improve the viscoelastic properties of these products, such as added hydrocolloids, cellulose, or malt, one or more of which are typically added to plant-based meat substitutes to enhance or improve the viscoelastic properties and the mouthfeel. Additionally, there is need for reducing the amounts of by-products that form when the high temperatures are used during the plant-based protein product processes. For example, acrylamides are known to form in food when temperatures during processing exceeds 120°C. Moreover, the environmental impact or the carbon footprint of the current processes that use high temperatures or high pressure could be further reduced.

[0003] For example, EP3166418B1 describes production of a protein product comprising a mixture of Vicia faba and pea protein flours at certain favorable ratios. The process uses temperatures in the range from 144°C to 147°C during the extrusion step that is necessary to form a desired consistency of the end-product.

[0004] FR 2340054 discloses a manufacturing method of textured vegetable proteins by mixing a paste containing 5-12 wt-% water and 95-88 wt-% of a leguminous flour having fatty content below 5 wt-%, vegetable protein content 55-75 wt-% and water content 8-12 wt-%. While maintained under pressure, the temperature of the paste is progressively raised to even higher temperatures that in the EP3166418B1, namely, 200-300 °C, while the water/flour mixture is held above 100 °C for less than 2 minutes. The product is then extruded through a die having one or more orifices.

[0005] GB 2132206 describes a smooth, elastic and even protein fibers that are formed by extruding a mixture of protein micellar mass (PMM) and added gellable starch into hot water. The starch is present in an amount up to about 30 wt-% of the PMM. The fibers are described to be useful in a variety of meat analog products. Temperatures during extrusion typically range between 150 to 260°C. [0006] CA 1075968 claims a method of texturizing a flour having a substantial starch containing legume content comprising the steps of forming a flour into a pumpable aqueous slurry. The slurry is pumpable at pressures of less than 200 pounds per square inch, requiring a significant amount of energy that generates a source of steam, separately passing the slurry and steam through a mixing jet and then into a cooler wherein said slurry and steam become intimately mixed and the flour becomes heated to at least 115 °C to texturize the slurry; and collecting and cooling the slurry after passage through the cooker; the aqueous slurry having at least 20 % protein (% nitrogen x 6.25).

SUMMARY

[0007] Provided are processes that produce legume-based meat substitutes wherein the process is performed at atmospheric pressure and during which the temperature of any of the process steps does not raise higher than 100°C. The legume-based meat substitutes produced by the process comprise only legume that has been fermented with a filamentous fungus and no other added ingredients, such as cellulose, hydrocolloids or malt, and yet, surprisingly, have viscoelastic properties and mouth-feel that closely resembles cooked meat. The product produced by the process further has substantially no process-caused by-products, such as acrylamides, because the temperature of the process does not, at any step of the process, exceed 100°C. The processes provided herein avoid the use of high pressure/high heat, such as extrusion techniques, that are often used in texturizing legume-based protein products to improve their mouthfeel and viscoelastic properties. Therefore, the process is also environmentally more sustainable than the processes the inventors are currently aware of.

[0008] The novel processes of transforming or texturizing legume raw material into a texture that resembles meat products provided herein rely on a process during which the temperature remains under 100°C during the entire plant-based protein/meat substitute manufacturing process. The process uses centrifugation of a fermented and heat-treated legume-product, that is performed at atmospheric pressure and at room temperature to texturize the legume-based meat substitute product. The inventor has surprisingly found that a process comprising the steps of (1) heating a fermented legume product at atmospheric pressure and at maximum 100°C after fermentation; (2) centrifuging at atmospheric pressure and room temperature after cooking; (3) settling the centrifuged product in a cool temperature after centrifugation, and (4) screening the settled product to remove loose bits and leaving the tight core; is sufficient to create texture and viscoelastic properties for pleasant, meat-resembling mouth-feel that further has a long shelf-life, without the need of additives such as added hydrocolloids or cellulose and without the use of an extruder. The shelf life can be up to about 30 days at cool temperature (at or under +6°C), and can be extended further to about 120 days at cool temperature (at or under +6°C), and can be further extended by heating the product after packaging/laminate-sealing, in a steam-oven at about 90°C for 40-90 min. The product can be irradiated or autoclaved after packaging to increase the storage life. Preferably, the product is irradiated. The product surprisingly also retains its elasticity after re-mixing in aqueous solutions or when defrosted after freezing. The starting material for the process is peeled, crushed and dried legume, rather than legume flour, which further reduces the energy needed for processing the legumes into the legume-based food product.

[0009] Prior to the centrifugation, the legume has been fermented by a filamentous fungus. In the first step of the process, water is added to the fermented product and the water/fermented product mixture is heat-treated at atmospheric pressure at no higher than 100°C, for example at 90-100°C, for example at 90-95°C for about 8-15 min. Surprisingly, the heat treatment was found to typically adjust the pH of the product to a range of between 4.5 and 5.2 without the need to use external pH adjusting agents, thus simplifying the process even further. The water is drained after the heat treatment and the legumes are cooled to under 40°C. The second step of the process is centrifugation to reduce the moisture by at least about 20%= followed by settling of the solids in a temperature at or under 6°C, for example at about 6°C, 5°C or 4°C or between 3-6°C. The solids can be optionally size-separated. The resulting product has a significantly improved mouthfeel, flavour and shelf-life compared to other legume-based food products, and can be vacuum-packed or even frozen without the risk of losing the viscoelastic properties achieved by the process. Moreover, because high heat or pressure are not used during the entire process, the process is much more environmentally friendly compared to products produced using legume flours and extruders in texturizing the products. Further, the lack of high heat during the process also results in a product that has substantially no high-heat induced potentially harmful byproducts, such as acrylamides.

[00010] Accordingly, provided is a process for improving composition and viscoelastic properties of plant-based protein products or meat substitutes characterized in that the process comprises of (1) subjecting a fermented legume culture to a heat-treatment, wherein the heat is performed at atmospheric pressure in temperature at no higher than 100°C; (2) subjecting the heat-treated legume to a centrifugation process at atmospheric pressure and at room temperature to reduce the moisture content of the fermented and heat-treated legume culture by at least 20%; (3) settling the centrifuged legume product for 12-24 hours at temperature under 6°C; and (4) screening the settled product to remove loose parts, thereby resulting in legume-based food product with improved composition and viscoelastic properties. The fermented heat-treated legume product does not contain texture-improving additives typically used in legume-based food products/meat alternatives, such as added hydrocolloids or added cellulose.

[00011] In some or all aspects of the process, the moisture reduction during the centrifugation process is between 20 and 40% for example by at least 25%, for example by at least 30%, for example by at least 35%, for example about 40%.

[00012] In some aspects, the screening is performed with a sieve having a mesh size of 7-12mm ² , for example, 8mm ² , 9mm ² , 10mm ² , for example 9mm ² , that results in separation of larger bits from smaller grits.

[00013] In some aspects, the screening comprises a second screening performed with a sieve having a mesh size of lmm ² -3mm ² , for example, about 1mm ² , 2mm ² , or 3 mm ² that separates the finer "flour" or "dust" from the bits and grits.

[00014] In some or all aspects of the process the heat-treatment is or has been performed at atmospheric pressure in temperature between 90°C and 100°C, for example 90°C-95°C. In one or all aspects of the process, the process further comprises the step of heat-treatment at atmospheric pressure in temperature between 90°C and 100°C, for example 90°C-95°C.

[00015] In some or all aspects of the invention the heat-treatment is or has been performed for 8- 15 min, for example 10-15 min, for example 10, 11, 12, 13, 14 or 15 min.

[00016] In some or all aspects of the invention, the process further comprises, prior to the step of centrifugation process, subjecting the fermented legume to the heat-treatment at atmospheric pressure in temperature between 90°C and 100°C, for example, at about 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C, or 100°C about 90°C-95°C for 8-15 min, for example 10-15 min, for example about 10, 11, 12, 13, 14 or 15 min. In some aspects the heat treatment is performed at 90°C-95°C for 10-15 min, for example, about 10, 11, 12, 13, 14, or 15 min.

[00017] In some or all aspects of the process after the centrifugation process the legume-based meat substitute product with improved composition and viscoelastic properties is capable of maintaining its viscoelastic properties when re-mixed in aqueous solution, and does not comprise additives, for example, cellulose, malt, or hydrocolloids, thereby resulting in a product with 0% externally added cellulose, malt, or hydrocolloids. The legume-based meat substitute product composition relies solely on the substances produced during the fermentation step.

[00018] In some or all aspects of the invention, the force used in the centrifugation process ranges between 31N and 38N. The centrifugation time can vary between about 60s and 150s. Typically, the greater the force, the shorter centrifugation time is needed to produce the desired consistency. For example, at force 34-36N, the centrifugation time is 120-130, for example, 115- 125 seconds, for example 120 seconds. In some or all aspects, the force is 31N and the centrifugation time is 150 seconds. In some or all aspects, the force is 34N and the centrifugation time is 120 seconds. In some or all aspects, the force is 32N, and the centrifugation time is 140 seconds. In some aspects, the force is 38N and the centrifugation time is 60 seconds. An example of a suitable centrifuge is Feltracon FE-800.

[00019] In one or all aspects of the process, the legume has been fermented with a filamentous fungus, for example, of Rhizopus oligosporus, Rhizopus oryzae, Rhizopus Stolonifer, Rhizopus arrhizus, Aspergillus spp., for example Aspergillus oryzae, or a mixture thereof. The legume has been peeled, crushed and dried prior to fermentation after which the dried product has been soaked in water and boiled for about 25-35 minutes, for example 30 minutes, and then cooled down to a cooled legume, to about 35-40°C, for example to about 35°C, 36°C, 37°C, 38°C, 39°C, or about 40°C which is used in the fermentation step. Once the fungus has been added to the cooled legume, fermentation time is typically, for example, for 24-48 hours, for example, about 24 hours, about 36 hours or about 48 hours. The temperature during fermentation is typically 25-37°C, for example 30-35°C. In some or all aspects of the process, the fungus is Rhizopus oligosporus. In some or all aspects of the process, the fungus is Aspergillus oryzae. In some or all aspects of the process, the fungus is Rhizopus oryzae. In some or all aspects of the process, the process further comprises the step of fermenting the legume with a filamentous fungus prior to the step of heattreatment and centrifugation process. In some or all aspects of the process, the process further comprises the soaking and boiling the legume prior to the step of fermentation. Examples of suitable fermentation conditions are provided, e.g., in Farnworth, E. 2008. Handbook of fermented functional foods. Second edition. CRC Press. New York, pp. 481-482.

[00020] In one or all aspects of the process the legume is selected from fava bean, yellow pea, soyabean, and any mixture thereof, for example fava bean and yellow pea; fava bean and soyabean; and yellow pea and soyabean; or fava bean, soyabean and yellow pea. In some or all aspects of the invention, the legume may also be selected from lentils or chickpeas or mixtures thereof, or the fava bean, yellow pea or soyabean may be combined with lentils or chickpeas or both. In one or all aspects of the process, the legume in the legume culture comprises over 90%- 100%, for example 95%-100%, for example 98%-100% peeled, crushed and dried legume. In one or all aspects of the process, the legume comprises 90-100% fava bean, preferably, 90-100% fava bean that has been peeled, crushed and dried, preferably, not ground to a legume-based flour. In one or all aspects of the process, the legume comprises 90-100% yellow pea. In one or all aspects of the process, the legume comprises 90-100% soyabean.

[00021] In one or all aspects of the process the legume has been peeled, crushed and dried before subjecting it to the fermentation process. In one or all aspects of the process, the process further comprises a step of peeling, crushing and/or drying the legume. Preferably, the legume is not legume flour.

[00022] In one or all aspects of the process the temperature does not exceed 100°C, for example it does not exceed 99°C, for example 98°C, for example 97°C, for example 96°C, for example 95°C, for example 94°C, for example 93°C, for example 92°C, for example 91°C, or for example 90°C. [00023] In all aspects of the process, the pressure is not altered from the atmospheric pressure. [00024] In one or all aspects of the process, the process comprises, prior to the centrifugation step, a further processing step of cutting or grinding the legume-based meat substitute product with improved composition and viscoelastic properties. Cutting can result in consistency and size of nuggets, grinding can result in consistency of ground meat.

[00025] In one or all aspects of the process further comprises vacuum packing the legume-based food product with improved composition and viscoelastic properties. The product can also be frozen. In one or all aspects of the process further comprises freezing the product either after centrifugation, after further processing, or after vacuum-packing.

[00026] In one or all aspects of the process, the process further comprises a step of heat-treating the fermented legume culture at atmospheric pressure at no higher than 90-100°C, for example no higher than 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C, or 100°C.

[00027] In one or all aspects of the process, the process further comprises a step of fermenting a peeled, crushed and dried legume product with a filamentous fungus prior to the heat-treatment. In some or all aspects, the fermentation is carried out for 24-48, for example 24 hours, 26 hours, 28 hours, 30 hours, 32 hours, 34 hours, 36 hours, 38 hours, 40 hours, 42 hours, 44 hours, 46 hours, or 48 hours. The fermentation temperature can be between 30°C and 35°C, such as about 30°C, 31°C, 32°C, 33°C, 34°C, or 35°C.

[00028]The improved composition produced by the process comprises substantially no byproducts that can be caused by or result from process steps that would include heating the legume-based food product to over 100°C, such by-products including, e.g., acrylamides that typically form in starchy vegetable products when they are heated to over 120°C temperatures. Such temperatures are typically reached if a process includes a high-pressure/high-heat extrusion step that is often used to improve the texture of a legume-based protein products, specifically products that use legume-based flour in the raw material mixture. Contrary to the typical processes used for texturizing legume-based protein products, extrusion is not used in any process step on the present process. Accordingly, provided is an improved composition prepared by the process of the invention wherein the composition comprises under 200 ppb of acrylamides, in some aspects the composition comprises under 150 ppb acrylamides and in some aspects the composition comprises under 100 ppb of acrylamides.

[00029] The product produced by the process preferably has hardness with an average peak force of about 1200g of 12N, as measured by Stable Microsystems TA.XT plus 100 C texture analyser.

[00030] In one aspect, provided is a legume product with improved viscoelastic properties and comprising substantially no high heat-induced by products, and which is capable of retaining its viscoelastic properties after it is mixed in water or if frozen, after defrosting, produced by any of the processes set forth herein.

BRIEF DESCRIPTION OF DRAWINGS

[00031] Figure 1 shows a basic flowchart of the PROCESS of the invention, comprising, the steps of (1) heating the fermented legume product at atmospheric pressure and at 90-100°C after fermentation to stop fermentation and to reduce acidity to pH between 4.3 and 5.3; (2) centrifuging after cooking to reduce the moisture content, wherein centrifugation is carried out in atmospheric pressure and at room temperature, i.e., at 20-22°C, using, for example, a force of 31- 38N for about 60-150 seconds after the acceleration, the centrifugation step reduces the moisture by at least 20%; (3) settling in a cold-room at +4-+6°C for 12-24 hours after centrifugation, to tighten the structure; (4) screening the settled product to remove loose bits and leaving the tight core. The process can further comprise one or more POST-PROCESSING steps, including, packing or vacuum-packing the product after screening and heat-treating with steam at about 90°C for 40-90 min, the vacuum-packed product to extend the shelf life to 120-150 days. Alternatively, the product can be autoclaved after packaging, which increases the storage time even further. The process can also comprise one or more PRE-PROCESSING steps, that may include one or more of (a) the legume is peeled, crushed and dried; (b) the dried product is soaked in water and boiled for about 25-30 minutes to reduce the amount of phytates, such as phytins, and vicines and convicines, and the boiled product is cooled down to 35-40°C, for example 38°C; (c) the fungus is added to the cooled legume and fermentation is carried out for 24-48 hours; (d) the fermented product is combined with water in an approximate ratio of one part of fermented product and two parts of water to assist in the heat-treatment; (e) prior to the centrifugation step, a step of additional processing can be added, wherein the fermented product can be further processed by vacuum packing and/or freezing and/or cutting, grinding or slicing. [00032] Figure 2 shows a deformation curve produced from the results for measuring hardness using a standard Stable Micro Systems food texture analyser. The texture analyser was used to quantitatively analyse the effect of the process to the mouthfeel of the end product compared to a product that was not processed using the process described herein. TP-t indicates a sample of "Tarhurinpapu tumma" (tumma=dark-colored product) and TP-v indicates a sample of "Tarhurinpapu vaalea" (vaalea=light-colored product). Force is shown on the y-axis, and time on the x-axis. The distance remains constant. The shaded area on the bottom shows a reference without resistance.

DESCRIPTION

[00033] Provided are improved processes to process legumes to produce an improved legumebased food product or legume-based meat alternative, also referred to herein as improved legume-based meat-alternative or legume-based protein product.

[00034] The inventor has surprisingly found that when peeled, crushed and dried legumes that have been heat-treated to reduce the amount of anti-nutrients, fermented with a filamentous fungus, and heat-treated at atmospheric pressure at no higher than 100°C to stop the fermentation and adjust the pH, a centrifugation process performed at room temperature and at atmospheric pressure that reduces the moisture content of the fermented product by at least 20%, is sufficient to produce an end-product that has viscoelastic properties and mouthfeel resembling meat. There is no need to add additional fillers, such as cellulose, malt or hydrocolloids, because the viscoelastic properties are surprisingly achieved by centrifugal drying of the fermented product alone. The added advantage of the process is that because the temperature during any step of the process does not reach higher than 100°C, at atmospheric pressure, the end-product contains no, or substantially no, high heat-induced by-products, such as acrylamides that form when a product is heated to 120°C or above. The process results in an end-product that has the elasticity and dryness that resembles a cooked meat product that can be used in food production as an alternative to meat, for example directly by a consumer or in industrial food production. A further advantage of the process is its lower environmental impact compared to known methods of producing plant-based meat alternatives, because it uses neither high pressure nor high temperatures.

[00035] The pre-processing, processing and post processing comprises the following steps: [00036] Pre-processing: Step (1): Soak the pealed crushed legume for 10-15 hours in one part legume, two parts water and adjust pH with approximately 2.5%-3.5%% acid, such as acetic acid, resulting in pH of 4.4-4.7, for example about 4.6. Drain. Step (2): add the same amount of water as in step (1) and add 1% acid, such as acetic acid (e.g., 100L water IL acetic acid) and boil for about 30 minutes. Drain and cool down to a temperature below 40°C, for example to 35-39°C. Step (3): Add starter culture mixing well. For example, one can use a starter that contains about 1- 2 million spores/lOOg of legume. Step (4): Ferment for 24-48 hours, maintaining fermenting temperature under 39°C. Optionally, the fermented product can be ground or cut into smaller sized, such as bite-sized, blocks at this stage.

[00037] Processing comprises the step of boiling the fermented product for example, for about 10 minutes to stop fermentation, and draining solids from the boiling water. Centrifuging. Cooling the solids to under 6°C, and letting them settle for 12-24 hours. Optionally, the settled solids can be size-separated, wherein the bits can be separated from the grits with, e.g., 9mm ² mesh size sieve; and grits can be separated from flour with, e.g., 2mm ² mesh size sieve.

[00038] Post-processing comprises packing, such as vacuum packing or laminate packing, pasteurization or irradiation or autoclaving. The product can be stored at 6°C or cooler.

[00039] Accordingly, in one embodiment, provided is a process for improving composition and viscoelastic properties of plant-based protein products or meat substitutes characterized in that the process comprises subjecting a fermented legume culture to the steps of boiling the fermented product to stop fermentation; centrifuging the boiled product after draining to reduce the moisture by at least about 20%; cooling the solids to under 6°C, and letting the cooled solids settle for 12-24 hours at under 6°C; and separating the settled solids based on the size into bits, grits and flour, wherein the centrifugation is performed at atmospheric pressure and at room temperature to reduce the moisture content of the fermented and heat-treated legume culture by at least about 20%, thereby resulting in legume-based food product with improved composition and viscoelastic properties.

Centrifugation

[00040] Accordingly, in one aspect, the process for improving viscoelastic properties of legumebased meat-substitute comprises a centrifuging process, wherein a fermented and heat-treated legume is subjected to centrifugation at atmospheric pressure and at room temperature to achieve moisture reduction by at least about 20%.

[00041] In some or all aspects of the process, the centrifugal force is at least about 31N, for example between about 31N and 38N, for example 31N, 32N, 33N, 34N, 35N, 36N, 37N or 38N, for example, about 34N.

[00042] The centrifugation time is between about 60 and 150 seconds, for example 80-120 seconds after the acceleration has resulted in the desired centrifugal force. A lower centrifugal force typically requires a longer centrifugation time. In some or all aspects of the invention, acceleration speed is at least about 3.4 m/s ² , for example at 3.4 m/s ² , and the centrifuging time, about 120s at about 34N force. An example of a suitable centrifuge is Feltracon FE-800, a skilled artisan can easily adjust alternative instruments for the purpose of centrifugation.

[00043] Centrifugation process is performed to achieve reduction in the moisture-content of the legume-based food product by at least 20%, for example by at least 25%, for example by at least 30%, for example by at least 35%, for example by at least 40%, compared to the moisture-content of the legume-based food product before the controlled centrifugation, and results in a legumebased food product with improved viscoelastic properties.

[00044] In some or all aspects of the process, the acceleration speed kept constant at about 3.4 m/s ² . How to adjust the acceleration speed to of a centrifuge to about 3.4 m/s ² will be dependent on the type of centrifuge and is well known to one of ordinary skill in the art. Too short centrifugation time with too little force will result in watery, soft and loose product, whereas the elastic and dry product can be achieved with sufficient force and time.

[00045] The acceleration force is dependent on the moisture content of the fermented and heat- treated, and drained legume product that is processed. Based on this description, one of ordinary skill in the art can adjust the force and time in such a manner that any centrifuge from laboratory, to pilot to industrial scale can be adjusted to perform the process with the information provided in this description.

[00046] Prior to the centrifugation process, the legume has been fermented by a filamentous fungus at a temperature, for example, between 30°C and 35°C for 24-48 hours, and heat-treated for about 8-15 min at atmospheric pressure at no higher than 100°C to stop the fermentation. [00047] The fermented legume can be cut into, for example, nuggets or ground to resemble ground meat or sliced prior to centrifugation so that the end-product, wherein the moisture has been reduced can resemble bite-sized nuggets, ground meat or meaty flour that can be used in making meat-like balls or patties. The product surprisingly maintains its viscoelastic properties, firmness and mouthfeel even after freezing and defrosting, as well as when it is mixed in aqueous solution, such as in a soup or a stew. That the product maintains its viscoelastic properties after re-mixed in water or after freezing and defrosting is unusual, because the currently available legume-based meat analogues/substitutes or alternatives lose their viscoelastic properties if added to a soup, or if frozen and defrosted, and deteriorate/disintegrate into a mushy or gel-like consistency. Legume

[00048] Essentially any legume can be used as a starting material for the process of the invention. Examples of legumes include fava bean, yellow pea, soyabean, lentils and chickpeas, and any mixtures thereof. In some aspects, the legume comprises over 50%, such as 60%, 70%, 80%, 90% or is 100% fava bean. In some aspects, the legume comprises over 50%, such as 60%, 70%, 80%, 90% or is 100% yellow pea. In some aspects, the legume comprises over 50%, such as 60%, 70%, 80%, 90% or is 100% soyabean. In some aspects, the legume is a mixture of fava bean and yellow pea. In some aspects, the mixture is yellow pea and soyabean. In some aspects the mixture is fava bean and soyabean.

[00049] The legume used in the process is pre-processed by peeling, crushing and drying prior to it being fermented. Preferably, the process does not use legume-based flours. In some or all aspects of the process, the process further comprises a step of peeling, crushing and/or drying the legume. It has been suggested that, for example, phytic acid, as well as vicins and convicins in plant foods complexes with dietary essential minerals such as calcium, zinc, iron, and magnesium and makes them biologically unavailable for absorption. Accordingly, these compounds have also been called "anti-nutrients." To reduce the amount of the anti-nutrients, such as phytins, vicins and convicins, the peeled, crushed and dried legume is soaked in water and boiled, prior to fermentation. The boiling typically takes 25-30 minutes and is performed in atmospheric pressure after which it is cooled to no higher than about 40°C, for example to room temperature of 20- 22°C. In some or all aspects of the process, the process further comprises a step of soaking the peeled, crushed and dried legume in water and boiling it. The typical soaking is performed using one part of legume and two parts of water and 3% acetic acid, or equivalent. After soaking, the water is drained off and equal amount of clean water and about 1% acetic acid adjusting the pH to under about 4.6, or equivalent, and the product is boiled for about 30 minutes after which the water is drained and the product is cooled to under 40°C, for example to about 39°C, about 38°C or about 37°C.

[00050] The legume product as used herein refers to a product made of legumes that have been peeled, crushed and dried, preferably not ground to a flour, and then subjected to fermentation with one or more filamentous fungus, and wherein after the fermentation, water is added to the fermentate, namely the entire product of the fermentation that has been processed in the fermentor has been heat-treated to stop the fermentation process.

[00051] The legume can be any legume. In some or all aspects of the process, the legume is selected from fava bean, soy bean and pea or a mixture thereof. [00052] In some or all aspects of the process, the legume comprises 85-100% fava bean, for example 90-100%, 95-100%, 98-100%, or consists of 100% of fava bean.

[00053] In some or all aspects on the process the legume is selected from fava bean, soy bean and pea or a mixture thereof.

Fermentation

[00054] Fermentation is carried during the pre-process. The legumes that have been cooled down in the earlier soaking-boiling steps are mixed with fungal spores or a starter culture and allowed to ferment typically for 24-48 hours at about 27°C -37°C, for example at 30-35°C in a fermenting chamber with sufficient constant moisture and oxygen. The legumes are naturally processed by the fungus during the fermentation. The fermentation techniques and conditions with filamentous fungi are well known (see, e.g., Farnworth, E. 2008. Handbook of fermented functional foods. Second edition. CRC Press. New York, for example at pp. 481-482).

[00055] The fungus is preferably selected from one of Rhizopus oligosporus, Rhizopus oryzae, Rhizopus Stolonifer, Rhizopus arrhizus, Aspergillus spp., for example Aspergillus oryzae. In some or all aspects of the process, the filamentous fungus is Aspergillus oryzae or Rhizopus oligosporus. Sometime, a mixture of fungi may be used, such as a mixture of Aspergillus oryzae and Rhizopus oligosporus. In some or all aspects of the process, the process further comprises a step of preprocessing comprising fermenting the peeled, crushed and dried legume with the filamentous fungus resulting in fermented legume product.

[00056] The filamentous fungus used in the fermentation can be selected from Rhizopus oligosporus, Rhizopus oryzae, Rhizopus Stolonifer, Rhizopus arrhizus, Aspergillus spp., for example Aspergillus oryzae, or a mixture thereof.

[00057] In some or all aspects on the process, the filamentous fungus is selected from the group consisting of Rhizopus oligosporus, Rhizopus oryzae, Rhizopus Stolonifer, Rhizopus arrhizus, Aspergillus spp., for example Aspergillus oryzae, and a mixture thereof.

[00058] In some or all aspects on the process, the Aspergillus spp, is Aspergillus oryzae.

[00059] In some or all aspects on the process, the filamentous fungus is a mixture of Aspergillus spp selected from Aspergillus oryzae and Aspergillus niger.

[00060] In some or all aspects on the process the filamentous fungus is Rhizobus oligosporus. [00061] In some or all aspects on the process the filamentous fungus is Aspergillus oryzae.

[00062] An example of a suitable fermentation protocol is set forth herein. Peeled and crushed legumes are soaked in water for 12-24 hours, for example 12-15 hours, for example about 12 hours, using approximately one-part legumes, two parts water and adjusting the pH with about 2.5%-3.5%, for example 3% acetic acid. The soaking water is discarded. About equal amount of water is added to the soaked, drained legumes and the combination of water and legumes are boiled for 10 minutes to 3 hours, for example for 20-40 minutes, for example about 20 min, 30 min, 40 min, or 50 min to kill any bacteria that may have grown during the soaking period and to break down anti-nutrients, such as phytates, such as phytins, and vicines and convicines, the pH is adjusted to under pH 4.6. After cooking, the water is drained. The legumes are cooled down to a temperature at about 35-40°C, for example to about 39°C or to 38°C or to 37°C. The fungal spores or a starter culture is mixed with the cooled legumes, at approximately at least 1 million spores per 100 legumes. Fermentation requires constant humidity, oxygen, and a constant temperature, which is typically 25-37°C, for example, at about 37°C. Thus, a fermentor made of stainless steel, wood or plastic is suitable when sufficient amount of air/oxygen can pass to the fermentation chamber to allow fermentation.

[00063] Temperature, length of fermentation and relative humidity are important for successful fermentation (Farnworth 2008, p. 480.). Increase in relative humidity speeds up the fermentation. Temperatures typically range between 25-37°C. The higher the temperature, the shorter the fermentation time. For example, the following has been suggested as examples for time when using different temperatures with 100g of boiled legumes and with about 1 million spores: [00064] 1. 25°C, 80 h

[00065] 2. 35-38°C 15-18 h [00066] 3. 25-37°C 20-50 h [00067] 4. 32°C 20-22 h

Heat-treatment

[00068] The purpose of the heat-treatment of the fermented product is to stop the fermentation. Accordingly, additional water is added to the fermented legume-product, such as fermented legume sheets, in a heating chamber, kettle, or pot, typically at ratio 1 part legume and 2 parts water (1:2 legume:water) so that it can be evenly heated. The heat-treatment is performed in atmospheric pressure and at no higher than 100°C, for example at 90°C -100°C, for example the temperature can be kept at about 90°C, 91°C, 92°C, 93°C, 94°C, 95°C, 96°C, 97°C, 98°C, 99°C or 100°C. The heat-treatment is applied for 8-15 min, for example 10-15 min, for example, about 10, 11, 12, 13, 14, or 15 min from the time the desired temperature, for example 90°C, is achieved at a heating chamber.

[00069] The heat-treatment also, surprisingly, results in adjustment of the pH of the heat-treated product to a range of 4.3 and 5.3, for example between pH 4.5-5.2. The fermented product prior to boiling is acidic as the fungus does not grow unless the pH of the fermenting culture is at or below 3.5, and the heat-treatment adjusts the pH slightly higher, i.e., pH 4.3-5.3, for example between pH 4.5-5.2. In some aspects, pH can be lowered closer to pH 4.3-4.5 with an acid, such as 1% acetic acid, to improve shelf-life.

[00070] After the heat treatment, the water is drained, and the drained legume product is subjected to optionally grinding or cutting or slicing, and then to centrifugation.

[00071] In some or all aspects of the process, the process if the invention comprises a step of heat-treating the fermented legume product prior to the centrifugation process.

Viscoelastic properties

[00072] An important attribute of real meat products is their ability to retain/lose fluids during cooking, as their fluid content impacts their look, feel, mouthfeel, and cooking properties. It is therefore important that meat analogs, such as legume-based meat-like protein products, simulate the fluid-holding properties of real meat, particularly real cooked meat. The fluid holding properties of meat analogs have been manipulated by altering biopolymer type, concentration, and crosslinking. In addition, the incorporation of polysaccharides has been used to improve the fluid holding properties. The processes provided herein rely on using filamentous fungus- fermented legume-product followed by a process that comprises heat-treatment of the fermented legumes, rapid centrifugation, a settling period in cool conditions, and screening that peels off loose parts after the settling is complete, to achieve the look, feel, mouthfeel, and cooking properties of the legume-based protein product or meat. The process does not use high heat, pressure or legume flour, and surprisingly produces a product that holds its properties both when cooked in aqueous environment as well as after freezing and defrosting, thereby resulting in a product that is significantly more versatile and, when heat-treated after vacuum packing, also allows longer storage time compared to other products, such as commercial plant-based meatlike products such as those from IMPOSSIBLE FOODS™ and BEYOND MEAT™, extending the storage for up to 150 days. Without heat treatment, the storage time of the product produced by the process described herein is about 30 days.

[00073] The term "meat-like legume-based protein product" as used herein refers to a food product that is not derived from an animal but has structure, texture, and/or other properties comparable to those of animal meat. The term refers to uncooked, cooking, and cooked meat-like food product unless otherwise indicated herein or clearly contradicted by context.

[00074] The term "mouth feel" as used herein refers to the overall appeal of a food product, which stems from the combination of several characteristics that together provide a satisfactory sensory experience. The mouth feel of a food product can be determined using a panel of human sensory experts.

[00075] The terms "Viscoelastic Properties" comprise "Texture Profile", which refers to the analysis of textural properties of a material by subjecting the material to a controlled force from which a deformation curve of its response is generated.

[00076] TP can be assessed using standard texture analyzers, for example, Stable Micro Systems texture analyzer.

[00077] For example, the hardness can be measured using Stable Microsystems TA.XT plus 100 C texture analyzer using 20-25mm ³ bits of the centrifuged fermented material that has been stored in under 6°C, preferably immediately after taking from the storage, and using 5.5g starting weights and at speed of 2mm/s. Using these conditions, it can be shown that compared to a product, that has not been processed using the process of the invention, is softer because the force needed to advance the measuring weight is smaller, and that in addition, the harder, processed product has a significantly smaller variation indicative of consistency of hardness in the process. The hardness values measured in the experiments include an average peak force of about 1200g of 12N. The bits collected with the sieve can be collected and used for, for example, making products that mimic meatballs or burger-like stakes. Figure 2 shows the measurements based on the analysis of hardness with Stable Microsystems TA.XT plus 100 C texture analyzer.

Composition

[00078] The composition of the product differs significantly from products that have been made from legume-based flours that have been texturized using additives such as cellulose, malt or hydrocolloids and extruders, because no added cellulose, malt or hydrocolloids are used in the texturizing process of the invention. Because no high heat has been used during the process, the product produced by the process of the invention also does not include heat-induced, and potentially harmful high heat-induced by-products such as acrylamides.

[00079] In some or all aspects of the method, the method further comprises a pre-processing step of cutting or grinding the fermented legume prior to centrifugation to produce the food product with improved viscoelastic properties.

[00080] In some or all aspects of the method, the method further comprises a pre-processing step of processing the fermented legume product through a meat grinder to result in legume-based food product with a cooked ground meat-like consistency. [00081] In some or all aspects of the method, the method further comprises a pre-processing step of cutting the fermented legume product prior to centrifugation into slices, nuggets, or other shapes resulting in a shaped legume-based product with a cooked meat-like consistency.

[00082] In some or all aspects of the method, the method further comprises a post-processing step of vacuum-packing the legume-based food product with improved viscoelastic properties or the product after processing.

[00083] In one embodiment, provided is a legume-based food-product with improved viscoelastic properties and composition, including an improved flavour profile, produced by the processes described herein. The product consists essentially of peeled, crushed dried legume that has been fermented with a filamentous fungus, wherein the fungus has been killed by a heat-treatment and wherein the heat-treatment has adjusted the pH of the heat-treated product to pH range of 4.3-5.3, and wherein the moisture has been reduced by at least about 20% using centrifugation, and wherein, optionally, the centrifuged product has been settled in under 6°C, and wherein, optionally, the solids have been size-separated resulting in a legume-based food product wherein the product resembles meat, such as cooked meat, for example, ground meat or a nugget, and wherein the product maintains its viscoelastic properties even when re-mixed in aqueous solution, such as a soup, or when defrosted after freezing. The product does not contain added hydrocolloids or cellulose and also contains substantially no by-products caused by process steps wherein heating over 100°C, such as over 120°C. An example of such by product is acrylamides that are a known harmful by-product in high-heat-treated processed plant-based products. In one aspect, the product produced by the contains less than 200 ppb, or less than 150 ppb, or less than 100 ppb of acrylamides.

[00084] The elastic and dry product of the process as described in this specification can further be processed using a "pre-processing step" that may include, for example cutting, slicing or grinding the fermented legume prior to centrifugation into ground product resembling cooked ground meat. Such pre-processed products can be used as a meat alternative either alone or in combination with other ingredients in industrial food production, or sold as is in retail for consumption by a consumer as a meat alternative/substitute/additive in various recipes. The step of cutting or grinding can be performed using any methods suitable for meat cutting, slicing or grinding known to a person skilled in the art.

[00085] The process may further comprise a post-processing step of packaging the product after settling or after size-separation. The packaging can comprise vacuum packaging or laminate packing. The packed product can be stored in cool temperatures of under 6°C or can be further be frozen. The vacuum-packed or laminate-packed product can be pasteurized, irradiated or autoclaved to increase storage time. Pasteurization and irradiation are preferred methods. It has been further discovered that, surprisingly, the frozen and subsequently defrosted product maintains its elastic properties that resemble cooked meat, and unlike other vegetable-based products, does not disintegrate into gel-like consistency.

[00086] Provided is also a product made by any of the processes described above. The product is substantially free of heat-induced by-products, such as acrylamides, which only form when a starchy food-product is exposed to heat over 120°C. The product further is substantially free of added cellulose or hydrocolloids.

[00087] Moreover, the combination of fermentation, short heat-treatment, and centrifugal moisture reduction followed by settling in cool temperature, the product in taste testing, has been judged as having a better taste profile compared to many other plant-based protein products currently on the market.

[00088] In this specification and the claims, which follow, reference will be made to a number of terms, which shall be defined to have the following meanings.

[00089] The term "room temperature" refers to temperature of 20-22°C.

[00090] Approximating language, as used herein and throughout the specification and claims, may be applied to modify any quantitative representation that could permissibly vary without resulting in a change in the basic function to which it is related. Accordingly, a value modified by a term or terms, such as "about”, is not to be limited to the precise value specified. In some instances, the approximating language may correspond to the precision of an instrument for measuring the value. The ratios given are weight ratios.

[00091] The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.

[00092] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description and some aspects of the invention includes instances where the event occurs and instances where it does not.

[00093] It is to be understood that the conditions described in the specification, figures and examples are only given as an example and that a person skilled in the art is readily able to find out optimal conditions for the process, when using other types of machinery.

[00094] The examples and comparative examples (commonly referred to as examples below) set forth herein further describe and demonstrate embodiments within the scope of the present invention. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations thereof are possible. EXAMPLES

Pre-processing the legumes

Fermentation

[00095] We based our fermentation protocol to that described in, e.g., a publication by Perttu Jokinen, published on 10 April 2013 by the Haaga-Helia University of Applied Sciences, entitled "Palkuainen Green Pea Tempeh in Vegetarian Food Recipes". Shortly, peeled and crushed legumes were soaked in water for 12-24 hours, using approximately one-part legumes, two parts water and adjusting the pH with 3% acetic acid. The soaking water was discarded. About equal amount of water was added to the soaked, drained legumes and the combination of water and legumes was boiled for about 20-40 minutes, to kill any bacteria that may have grown during the soaking period and to break down anti-nutrients, such as phytates, such as phytins, and vicines and convicines. After cooking, the water was drained. The legumes were cooled down to a temperature under 40°C. The fungal spores or a starter culture is mixed with the cooled legumes, at approximately at least 1 million spores per 100 legumes. Fermentation requires constant humidity, oxygen, and a constant temperature, which is typically 25-37°C, for example, at about 37°C. Thus, a fermentor made of stainless steel, wood or plastic is suitable when sufficient amount of air/oxygen can pass to the fermentation chamber to allow fermentation.

[00096] Temperature, length of fermentation and relative humidity are important for successful fermentation (Farnworth 2008, p. 480.). Increase in relative humidity speeds up the fermentation. Temperatures typically range between 25-37°C. The higher the temperature, the shorter the fermentation time.

[00097] Most of our experimentation was performed using pealed crushed and dried fava beans that were soaked for about 10-15 hours about one-part legumes, two parts water and 3% acetic acid to adjust the pH in the range of 4.3-4.8. The soaking water was removed and an equal amount to the original amount of water was added along with 1% acetic acid and the product is boiled for about 20-40 min, such as 30 min, after which the boiling water was drained and the pre-processed legumes are cooled to a temperature of under 40°C.

[00098] We added the starter fungus into the cooled legumes and mixed it thoroughly. The product was then processed through a meat grinder using a blade with holes of 2.5 mm of diameter, and laced in a chamber that was covered with a sheet, which was covered with a lid to allow sufficient air to enter the chamber for the fermentation process. Processing

Heat treatment

[00099] The fermented product was mixed with water and heated at atmospheric pressure to temperatures between 90°C and 100°C, typically for about 8-15 min, usually between 10 and 15 minutes resulting in a heat-treated "fermentate" or fermented legume product.

[000100] The pH was measured after the heat-treatment, and was consistently found to range between 4.3 and 5.3, usually between 4.5 and 5.2 without addition of any external pH adjusting agents. The pH increased from the original pH used during the fermentation thus resulting in mild and pleasurable flavor profile of the product.

Centrifugation

[000101] The heat-treated fermentate was subjected to a centrifugation at atmospheric pressure and room temperature to remove excess water.

[000102] With our industrial capacity Feltracon FE-800 centrifuge, we found that once a force between 31N and 38N was achieved, and the force was maintained for about 60-150 seconds, we surprisingly obtained a product, which had the elasticity, consistency, and mouth-feel of meat product. When we ground the fermented sheets prior to centrifugation, the end-product resembled ground meat; when we cut it, it resembled chicken nuggets. Prior to moving to the industrial capacity centrifuge, we also successfully used smaller centrifuges. The centrifugal force applied to product is an important factor to obtain most pleasant structure of end product.

Settling

[000103] The centrifuged product was allowed to settle in a cool room at under 6°C for 12- 24 hours which resulted in tightening of the structure of the legume product.

Screening (optional)

[000104] The settled product was screened to size-separate the differently sized solids into bits, grits and "flour" or "dust" which is smaller than grits, typically smaller than 2mm ³ , or to peel off any loose material leaving behind the tight core material that had been settled on the bottom during the settling step. In one experiment, we performed the screening using a first sieve with mesh size about 9mm ² to remove the larger loose bits and a second screening using a mesh size 2mm ² . Different mesh size can be used according to the desired size of the end-product. Viscoelastic properties

[000105] After the process, the product had a pleasant taste and mouthfeel of hardness and chewiness as assessed by humans. The product closely resembled cooked meat in its consistency. We were also surprised that unlike other plant-based meat substitute products, which typically disintegrate to a slimy consistency when re-mixed in water, the product we obtained after the process was firm and stayed together even when re-mixed in aqueous soups. Moreover, the product maintained its viscoelastic properties and mouthfeel when defrosted after freezing.

[000106] In addition to human taste and mouthfeel testing, we measured the viscoelastic properties of the product produced by the process using the Stable Microsystems texture analyzer.

[000107] The texture analyzer was used to analyze the mouthfeel of the product before the process and after the process thus imparting us knowledge of the effect of the process. The texture analyzer can be used to measure at least firmness/toughness; stiffness; cutting strength; toughness; and tensile strength. For example, we measured the hardness of the product using Stable Microsystems TA.XT plus 100 C texture analyzer.

[000108] The frequency of the sampling was 500 measurements per second using 5.5 g starting weight and initial speed of 2 mm/s. The analyzer analyses the force needed to advance the measuring head through a constant distance. This provides a table from which one can draw the graph shown in Figure 2. The results show that a sample that has not gone through the process step needs less force to advance the constant distance thus indicating that it is softer than the product that has been processed using the process of heat-treatment, centrifugation, settling and filtering/sieving.

[000109] The samples were taken from products that were stored at under 6°C. The average size of the samples was 20-25 mm ³ and they were measured immediately after taking them from the cool storage. We measured five parallel samples from both non-processed and processed products, and the analyzer automatically calculated the mean, standard deviation and variation (see table 2).

[000110] The results show that the processed samples had less variation, and thus the processed products were more consistent. Composition

[000111] Unlike other plant-based meat substitutes which typically use hydrocolloids or cellulose to improve the mouthfeel, or firmness or elasticity, our novel process does not use additives other than the fungus in the pre-processing fermentation step. [000112] It is well-known that potentially harmful by-products, such as acrylamides, are formed, when starchy food is processed at temperatures that reach 120°C or higher. The temperatures used in the currently known plant-based meat substitute processes appear to be well over 120°C.

[000113] Our process completely eliminates this potential problem because the temperature during the entire production process never reaches higher than 100°C.

000114] Table 1 below sets forth the results from the Stable Micro Systems texture analyzer.

Table 1

Start Batch Tarhurinpapu_tumma Tarhurinpapu_tumma

Tarhurinpapu_tumma01 Tarhurinpapu_tumma 1530.914 2380.314

Tarhurinpapu_tumma02 Tarhurinpapu_tumma 1659.821 2212.032

Tarhurinpapu_tumma03 Tarhurinpapu_tumma 1177.01 1632.258

Tarhurinpapu_tumma04 Tarhurinpapu_tumma 1129.458 1708.982

Tarhurinpapu_tumma05 Tarhurinpapu_tumma 825.824 1267.859

End Batch Tarhurinpapu_tumma Tarhurinpapu_tumma

Average: Tarhurinpapu_tumma (F) AVERAGEf'BATCH") 1264.605 1840.289

S.D. Tarhurinpapu_tumma (F) STDEVf'BATCH") 333.76 452.221

Coef. of Variation Tarhurinpapu_tumma (F) STDEV("BATCH") / AVERAGE("BATCH") * 100 26.392 24.573

Start Batch Tarhurinpapu_vaalea Tarhurinpapu_vaalea

Tarhurinpapu_vaalea01 Tarhurinpapu_vaalea 353.366 580.722

Tarhurinpapu_vaalea02 Tarhurinpapu_vaalea 890.013 904.069

Tarhurinpapu_vaalea03 Tarhurinpapu_vaalea 759.353 1063.319

Tarhurinpapu_vaalea04 Tarhurinpapu_vaalea 751.687 1033.507

Tarhurinpapu_vaalea05 Tarhurinpapu_vaalea 289.298 314.519

End Batch Tarhurinpapu_vaalea Tarhurinpapu_vaalea

Average: Tarhurinpapu_vaalea (F) AVERAGEf'BATCH") 608.744 TI . I

S.D. Tarhurinpapu_vaalea (F) STDEVf'BATCH") 269.022 322.611

Coef. of Variation Tarhurinpapu_vaalea (F) STDEV("BATCH") / AVERAGE("BATCH") * 100 44.193 41.401

000115] Table 2 sets forth average force measurements using the Stable Micro Systems texture analyser.

Table 2 g

Peak Force (Cycle: 1)

Test ID Batch Peak Force (Cycle: 1)

Start Batch Tarhurinpapu_tumma Tarhurinpapu_tumma

Average: Tarhurinpapu_tumma (F) AVERAGEf'BATCH") 1264.605

S.D. Tarhurinpapu_tumma (F) STDEVf'BATCH") 333.76

STDEVf’BATCH") /

Coef. of Variation Tarhurinpapu_tumma (F) AVERAGEf'BATCH") * 100 26.392

Start Batch Tarhurinpapu_vaalea Tarhurinpapu vaalea

Average: Tarhurinpapu_vaalea (F) AVERAGEf'BATCH") 608.744

S.D. Tarhurinpapu_vaalea (F) STDEVf'BATCH") 269.022

STDEVf’BATCH") /

Coef. of Variation Tarhurinpapu_vaalea (F) AVERAGEf'BATCH") * 100 44.193