WO2021195175A1

WO2021195175A1 - Fibrous meats by solid-state fermentation and extrusion

Info

Publication number: WO2021195175A1
Application number: PCT/US2021/023824
Authority: WO
Inventors: Aleh Manchuliantsau
Original assignee: Usarium Inc
Priority date: 2020-03-24
Filing date: 2021-03-24
Publication date: 2021-09-30

Abstract

A method, where fibrous meat analogs manufactured from plant-based ingredients, characterized in that the raw ingredients were subjected to extrusion to create a porous structure, then solid state fermentation in the presence of fungi to convert carbohydrates into additional protein and second extrusion, which works as a kill step and creates a fibrous texture similar to meat. The method reduces by-products, water consumption, avoids chemical treatment, improves protein quality and cut costs for meat analogs manufacturing.

Description

TITLE

FIBROUS MEATS BY SOLID-STATE FERMENTATION AND EXTRUSION

CROSS-REFERENCE

[0001] This application claims priority to U.S. Provisional Patent Application No.

62/993,746, which was filed on Mar. 24, 2020, U.S. Provisional Patent Application No. 63/017,947, which was filed on Apr. 30, 2020, U.S. Provisional Patent Application No. 63/076,426, which was filed on Sep. 10, 2020, and U.S. Provisional Patent Application No. 63/113,262, which was filed on Nov. 13, 2020. The contents of which are incorporated by entirety into this specification.

FIELD

[0002] The present disclosure relates to methods of manufacturing meat-like products with fibrous texture.

BACKGROUND

[0003] Fermented plant products such as Tempeh have texture and taste which may be undesirable for consumers looking for meat substitution. Fibrous meats made from plants by extrusion cooking process gain popularity, but require expensive ingredients. New, cost-effective methods of manufacturing meat-analogs are required.

SUMMARY

[0004] According to the present disclosure, a method is provided. The method, where fibrous meat analogs manufactured from oilcakes, spent grains, distillers dry grains, pulses, etc., characterized in that the raw ingredients were subjected to extrusion to create a porous structure, then solid state fermentation in the presence of fungi to convert carbohydrates into additional protein and second extrusion, which works as a kill step and creates a fibrous texture similar to meat.

[0005] It is understood that the inventions disclosed and described in this specification are not limited to the aspects summarized in this Summary. The reader will appreciate the foregoing details, as well as others, upon considering the following detailed description of various non-limiting and non-exhaustive aspects according to this disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

[0006] The features and advantages of the examples, and the manner of attaining them, will become more apparent, and the examples will be better understood, by reference to the following description of embodiments taken in conjunction with the accompanying drawings, wherein:

[0007] FIG. l is a process flow diagram of a method to process raw ingredients into a fibrous meat-like product according to Example #1 of the present disclosure;

[0008] FIG. 2A is an image of meat analog according to Example 1 of the present disclosure;

[0009] FIG. 2B is an image of meat analog according to Example 2 of the present disclosure;

[0010] FIG. 2C is an image of meat analog according to Example 3 of the present disclosure;

[0011] Corresponding reference characters indicate corresponding parts throughout the several views. The exemplifications set out herein illustrate certain embodiments, in one form, and such exemplifications are not to be construed as limiting the scope of the appended claims in any manner.

DETAILED DESCRIPTION [0012] Various examples are described and illustrated herein to provide an overall understanding of the structure, function, and use of the disclosed methods, systems, compositions, and products. The various examples described and illustrated herein are non- limiting and non-exhaustive. Thus, the invention is not limited by the description of the various non-limiting and non-exhaustive examples disclosed herein. Rather, the invention is defined solely by the claims. The features and characteristics illustrated and/or described in connection with various examples may be combined with the features and characteristics of other examples. Such modifications and variations are intended to be included within the scope of this specification. As such, the claims may be amended to recite any features or characteristics expressly or inherently described in, or otherwise expressly or inherently supported by, this specification. Further, Applicant reserves the right to amend the claims to affirmatively disclaim features or characteristics that may be present in the prior art. The various examples disclosed and described in this specification can comprise, consist of, or consist essentially of the features and characteristics as variously described herein.

[0013] Any patent, publication, or other disclosure material identified herein is incorporated herein by reference in its entirety unless otherwise indicated but only to the extent that the incorporated material does not conflict with existing definitions, statements, or other disclosure material expressly set forth in this specification. As such, and to the extent necessary, the express disclosure as set forth in this specification supersedes any conflicting material incorporated by reference herein. Any material, or portion thereof, that is said to be incorporated by reference into this specification, but which conflicts with existing definitions, statements, or other disclosure material set forth herein, is only incorporated to the extent that no conflict arises between that incorporated material and the existing disclosure material. Applicant reserves the right to amend this specification to expressly recite any subject matter, or portion thereof, incorporated by reference herein.

[0014] Reference throughout the specification to “various examples,” “some examples,”

“one example,” or “an example”, or the like, means that a particular feature, structure, or characteristic described in connection with the example is included in at least one example.

Thus, appearances of the phrases “in various examples,” “in some examples,” “in one example”, or “in an example”, or the like, in places throughout the specification are not necessarily all referring to the same example. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more examples. Thus, the particular features, structures, or characteristics illustrated or described in connection with one example may be combined, in whole or in part, with the features structures, or characteristics of one or more other examples without limitation. Such modifications and variations are intended to be included within the scope of the present examples.

[0015] In this specification, unless otherwise indicated, all numerical parameters are to be understood as being prefaced and modified in all instances by the term “about,” in which the numerical parameters possess the inherent variability characteristic of the underlying measurement techniques used to determine the numerical value of the parameter. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical parameter described herein should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0016] All ranges recited herein are inclusive of the end points of the recited ranges. For example, a range of “1 to 10” includes the end points 1 and 10. Also, any numerical range recited herein includes all sub-ranges subsumed within the recited range. For example, a range of “1 to 10” includes all sub-ranges between (and including) the recited minimum value of 1 and the recited maximum value of 10, that is, having a minimum value equal to or greater than 1 and a maximum value equal to or less than 10. Any maximum numerical limitation recited in this specification is intended to include all lower numerical limitations subsumed therein and any minimum numerical limitation recited in this specification is intended to include all higher numerical limitations subsumed therein. Accordingly, Applicant reserves the right to amend this specification, including the claims, to expressly recite any sub-range subsumed within the ranges expressly recited. All such ranges are inherently described in this specification.

[0017] The grammatical articles “a,” “an,” and “the,” as used herein, are intended to include “at least one” or “one or more,” unless otherwise indicated, even if “at least one” or “one or more” is expressly used in certain instances. Thus, the foregoing grammatical articles are used herein to refer to one or more than one (i.e., to “at least one”) of the particular identified elements. Further, the use of a singular noun includes the plural, and the use of a plural noun includes the singular, unless the context of the usage requires otherwise.

[0018] In this specification, unless otherwise indicated, all percentages (e.g., weight percent protein, percent protein, percent moisture) are to be understood as being based on weight.

[0019] There is a desire for alternative sources of protein that do not come from animal meat. Raw materials, such as, for example, beans, have been of interest as a protein source.

Solid state fermentation with fungi, such as, for example, the Tempeh process, may be used to process the beans for consumption. The Tempeh process is a natural fermentation process involving a Rhizopus fungus. Although the Tempeh process is clean, the resulting biomass from the Tempeh process can be characterized as having randomly oriented filaments on the chunks of raw ingredients with a texture, and a taste different from meatpacking muscle-like fibers), which may be undesirable for consumers looking for a meat substitution.

[0020] The grown mycelium can be separated (see US20200024577A1, 2018 Ecovative

Design process) from its medium for further texture improvement by 3D shaping and extruding (see Also 20200093167, 20200093155, 2018, Better Meat Co process), but harvesting and separation from the medium are costly.

[0021] Fibrous meat analogs can comprise at least one of a textured protein, a high moisture meat analog (HMMA), or a texturized vegetable protein (TVP), or any combinations thereof. HMMA are typically produced by high moisture extrusion cooking and can have a high final moisture content, such as, for example, 50 percent to 80 percent moisture by total weight of the HMMA. Some HMMAs may have a limited shelf life and require cold storage. TVP is typically produced by a lower moisture process than HMMA (e.g., less than 30 percent added moisture) and dried down to 8 percent moisture based on the total weight of the TVP. TVPs are typically marketed as a shelf stable product which requires rehydration before consumption. HMMA can be produced from TVP by adding additional moisture.

[0022] Fibrous meat analogs are made from chemically processed with alkali, acids and alcohols protein isolates and concentrates, which causes critique from healthy customers. Protein concentration and isolation processes are energy-intensive, require up to 301 of water for 1 kg of ingredient, lose up to 25% protein, and increase the cost per gram of protein up to 70% compared to raw ingredients.

[0023] An example of low water, low by-product processing of meat analogs is provided in U.S. Patent No. 10,645,950 where ingredients subjected to protein de-solubilization by extrusion, followed by washing impurities with water and extrusion into high moisture meat analogs. Presented process reduces protein loss down to 9-16%, but still wastes soluble carbohydrates during washing.

[0024] Additionally, the present inventor has determined the production of animal meat has a second role in preservation of the planet that is not as well known. For example, reducing the production of animal meat may result in a reduction in carbon dioxide emissions and water use. However, there are concerns with what to do with the by-product feed materials currently used in the production of animal meat. Burning or composting the by-products can cause greenhouse gas and methane emissions similar to animal meat production. Protein isolation from by-products can be costly due to low protein levels and protein isolation can generate new byproducts. Additionally, liquid fermentation may not be applicable to solid by-products.

[0025] Goal of this invention is to reduce by-products down to zero, reduce water consumption down to 0.5 liters per kg of the final product, avoid chemical treatment, improve protein quality, cut costs and deliver meat analogs with fibrous meat-like texture.

[0026] The foregoing goals are accomplished by a novel method, where fibrous meat analogs manufactured from oilcakes, spent grains, distillers dry grains, pulses, etc., characterized in that the raw ingredients were subjected to extrusion to create a porous structure, then solid- state fermentation in the presence of fungi to convert carbohydrates into protein and the second extrusion which works as a kill step and creates a fibrous texture similar to meat.

[0027] In examples according to the present disclosure the method for meat-like fibrous structure manufacturing comprises raw ingredients subjecting to extrusion at a temperature 130°C, a pressure 15 bar, and a speed in a range of 350 RPM to create a porous structure; solid state fermentation of the extruded ingredients in the presence of filamentous fungi for 48 hours, at a moisture content of 30% by total weight of the raw ingredients and temperature of 25-30°C to convert carbohydrates into protein; subjecting the fermented ingredients to extrusion at a temperature 150°C, a pressure 15 bar, and a screw speed of 350 RPM to accomplish a kill step and create a fibrous texture similar to meat.

[0028] The texture, the amount of by-products generated, water consumption, presence of chemical treatment, protein quality (PDCAAS), capital needs per typical line and costs are improved by disclosed invention compared to known methods of manufacturing of alternative meat made by Tempeh process, by extrusion from isolates and concentrates, made by desolubilization and washing and extrusion, and by separated and shaped/extruded mycelium as shown in the Table 1 below.

[0029] Table 1. Comparison of known methods (alternative meat by Tempeh process, alternative meat made from isolates and concentrates, alternative meat made by de-solubilization and washing, alternative meat by separated and shaped/extruded mycelium) to alternative meat made by the offered invention

[0030] Alternative meat manufacturing by extrusion requires high, preferably 55% by dry weight protein content. Since crops and by-products are naturally lower in protein than required, it has been accepted by those skilled in this art that in order to increase the protein content you need to separate proteinous from non-proteinous fractions by one or another method, which often results in by-products with limited market value.

[0031] In the course of the research leading up to the present invention, it was unexpectedly found that solid-state fermentation of raw ingredients in the presence of fungi helped to convert carbohydrates into additional protein and increase the total protein content to the levels high enough for successful extrusion into a fibrous texture similar to meat, with a chewy texture and without generating by-products. Low water consumption and the absence of by-products further reduced the costs.

[0032] The protein content comparison before and after solid-state fermentation is provided in the Table 2 below.

[0033] Table 2. Protein content comparison before and after solid-state fermentation

[0034] Protein digestibility-corrected amino acid score - a method of evaluating the quality of a protein-based on both the amino acid requirements of humans and their ability to digest it - used for ranking protein sources by their quality. Extrusion cooking is known to increase the digestibility of the protein for human consumption, but it has been accepted by those skilled in this art that amino acids content per gram of protein during various concentration and/or isolation processes remains the same.

[0035] In the course of the research leading up to the present invention, it was unexpectedly found that solid-state fermentation of raw ingredients in the presence of fungi helped to increase methionine by 8.7% (the most limiting amino acid of soybeans), along with other amino acids increasing overall protein quality calculated by PDCAAS method. We attribute this effect to the process where Fungi digesting sugars in the raw ingredient generate enzymes that break down complex carbohydrates into simple carbohydrates, digesting these simple carbohydrates further fungi produce extra protein with increased amounts of amino acids as shown in the Table 3 below.

[0036] The results of Soybean meal Amino Acid analyses before and after solid-state fermentation are provided in the Table 3 below.

[0037] Table 3. Soybean meal Amino Acid profile before and after solid-state fermentation, mg/g of protein.

[0038] The raw ingredients utilized in the process can be chosen from a non-limiting variety of oilseeds, spent grains, legumes, grains, starchy vegetables, algae and mixes thereof. For example:

• the oilseed byproducts comprising at least one of a sunflower oilcake, a soybean oilcake, a cottonseed oilcake, a rapeseed oilcake, a canola oilcake, a copra oilcake, a palm kernel oilcake, a peanut oilcake, or a combination thereof;

• raw ingredients comprising at least one of distillers dried grains with solubles, brewers spent grain, fruit and vegetable pomaces, bran, legumes, algae or a combination thereof;

• raw ingredients further comprising supplemented nitrogen, minerals, and vitamins from a mineral source, an organic source, a waste water source, or a combination thereof.

[0039] During solid state fermentation the heat transfer through the biomass can be challenging which impacts the yield. Agitation of solid biomass is labor-intensive and breaks filaments of the fungi, which again impacts the yield. The present inventors found utilizing a first extrusion allows to create pores for airflow, resolve the problem with overheating and remove agitation. Additionally, the extrusion can improve the digestibility of starches for fungi.

[0040] First extrusion parameters may be:

• a moisture in the barrel: in a range of 5-40wt% such as, for example, 10-17wt%. In certain examples, too few or too much moisture may lead to less expansion.

• a temperature in a range of 90-200C, such as, for example, 140C tol60C. In certain examples, the temperature may be 150C. In certain examples, a temperature too low may not puff/expand properly and too high may burn the ingredients.

• a pressure in a range of 1-150 bar, such as, for example, 70 bar to 90 bar. In various examples, the pressure is 80 bar. A pressure too low may not puff/expand properly and a pressure too high may explode.

• a speed in a range of 80-400 RPM, such as, for example 300 RPM to 400RPM. In various examples, the speed may be 350RPM. In various examples, the speed can influences the residence time within the extruder and you may need to lower it to cook the ingredients properly and utilize a higher speed to increase the throughput

[0041] The above parameters can inter-influence each other and an equilibrium of the parameters may be found for optimizing pores needed with the desired throughput. The first extrusion can be set to produce a pore size of at least 0.001 mm in the extrudate after the first extrusion prior to SSF, such as, for example, at least 0.01mm, at least 0.1mm, at least 1 mm, at least 2mm, at least 3mm, at least 4mm, at least 5mm, at least 6mm, at least 7mm, at least 8mm, at least 9mm, at least 10mm, at least 1 lm, at least 12mm, at least 13mm, at least 14mm, at least 15mm, at least 16mm, at least 17mm, at least 18mm, or at least 19mm. In various examples, the pore size can be no greater than 20mm, such as, for example, no greater than 19mm, no greater than 18mm, no greater than 17mm, no greater than 16mm, no greater than 15mm, no greater than 14mm, no greater than 13 mm, no greater than 12mm, no greater than 11mm, no greater than 10mm, no greater than 9mm, no greater than 8mm, no greater than 7mm, no greater than 6mm, no greater than 5mm, no greater than 4mm, no greater than 3mm, no greater than 2mm, no greater than 1mm, no greater than 1mm, or no greater than 0.1mm. For example, the pore size in the extrudate after the first extrusion may be in a range of 0.001mm to 20 mm, such as, for example, 0.01mm to 20 mm, 1 mm to 20 mm, 1 mm to 10 mm, 2 mm to 8 mm, 3 mm to 7 mm, or 4 mm to 6mm. In various examples, the pore size in the extrudate after the first extrusion can be 5mm. In some examples, the extrudate after the first extrusion can comprise a pore size at least 0.001mm up to 1.999 mm, such as, for example, 0.2mm. The pore size may be lowered to less than 0.01 mm and to compensate for the lower pore size air pumped at pressure may be added to the SSF. However, the pressurized air can damage fungi. Additional starch may be added to the ingredients to increase the pore size. For example, the raw ingredients may contain starch up to 80% of dry weight for better expansion, such as, for example a starch content in a range of 50% to 80% of dry weight.

[0042] Different substrates may require specific fungi for optimal solid state fermentation. In addition to Aspergillus oryzae used in the examples shown below, the Fungi used for solid state fermentation (SSF) can be chosen from:

• Fungi is from the phylum Basidiomycota, Ascomycota, Glomeromycota, Mucoromycota, or Zoopagomycota.

• Fungi is from the division agaricomycotina, ustilagomycotina, pezizomycotina, saccharomycotina, taphrinomycetes, diversisporalis, archaeosporales, paraglomerales, endogonales, mucorales, mortieralles, entomophthoromycotina, asellariales, kickxellales, dimargaritales, harpellales, zoopagomycotina, or combinations thereof.

• Fungi is from the class tremellomycetes, dacrymycetes, agaricomycetes, exobasisiomycetes, ustilaginomycetes, malasseziomycetes, moniliellomycetes, arthoniomycetes, coniocybomycetes, dothideomycetes, eurotiomyctes, geoglossomycetes, laboulbeniomycetes, lecanoromycetes, leotiomycetes, lichinomycetes, orbiliomycetes, pezizomycetes, sordariomycetes, xylonomycetes, or combinations thereof. • Fungi is from the order filobasidiales, agaricales, amylocorticiales, atheliales, boletalesjaapiales, lepidostromatales, geastrales, gomphales, hysterangiales, phallales, auriculariales, cantherellales, corticiales, gleophylalles, hymenochaetales, polyporales, russulales, sebacinales, stereopsidales, thelephorales, trechisporales, ceraceosorales, doassansiales, entyomatales, exobasidiales, georgefischeriales, microstromatales, tilletiales, urocystales, ustilaginales, malassezioales, moniliellales, saccharomycetales, coronophorales, glomeralles, hypocreales, melanosporales, microascales, boliniales, calosphaeriales, chaetospheriales, coniochaetales, diasporthales, magnaporthales, ophiostomatales, sordariales, xylariales, koralionastetales, lulworthiales, meliolales, phylachoralles, trichosphariales, eurotiales, chaetothyriales, pyrenulales, verrucariales, onygenales, mortierellales, mucorales, endogonales, or combinations thereof.

• Fungi is from the family Filobasidium, Dacromycetaceae, Agaricaceae, Amanitaceae, Bolbitiaceae, Broomeiceae, Chromocyphellaceae, Clavariaceae, Cortinariaceae, Cyphellaceae, Enolomataceae, Fistulinaceae, Himigasteraceae, Hydnangiaceae, Hygrophoraceae, Inocybaceae, Limnoperdacea, Lyophyllaceae, Marasmiaceae, Mycenacea, Niaceae, Pellorinaceae, Physalacriaceae, Pleurotacea, Pluteaceae, Porotheleaceae, Psathyrellaceae, Pterulacea, Schizophyllaceae, Stephanosporaceae, Strophariaceae, Tricholomataceae, Typhulaceae, Boletaceae, Boletinellaceae, Coniophoraceae, Diplocystaceae, Gasterellaceae, Gastrosporiaceae, Gomphidiaceae, Gyroporaceae, Hygrophoropsidaceae, Paxillaceae, Protogastraceae, Rhizopogonaceae, Sclerodermataceae, Serpulaceae, Suillaceae, Tapinellaceae, Hymenochaetaceae, Repetobasidiaceae, Schizoporaceae, Cystostereaceae, Fomitopsidaceae, Fragiporiaceae, Ganodermataceae, Gelatoporaceae, Meripilaceae, Merulaciaea, Phenerochaetaceae, Polyporaceae, Sparassidaceae, Steccherinaceae, Xenasmataceae, Albatrellaceae, Amylostereaceae, Auriscalpaceae, Bondarzewiaceae, Echinodontiaceae, Hericiaceae, Hybogasteraceae, Lachnocladiaceae, Peniphoraceae, Russulaceae, Gloeocyctidiellacceae, Stereaceae, Ustilaginomycetes, Saccharomycetaceae, Saccharomycodaceae, Saccharomycopsidaceae, Chaetomiaceae, Lasiosphaeriaceae, Sordariaceae, or any combination thereof.

• Fungi is from the genus Neurospora, Aspergillus, Trichoderma, Pleurotus, Ganoderma, Inonotus, Cordyceps, Ustilago, Rhizopus, Tuber, Fusarium, Pennicillium, Xylaria, Trametes, or any combination thereof.

• Fungi is from the species Aspergillus oryzae, Rhizopus oryzae, Fusarium graminareum, Cordyceps militaris, Cordyceps sinensis, Tuber melanosporum, Tuber magnatum, Pennicillium camemberti, Neurospora intermedia, Neurospora sitophila, Xylaria hypoxion, or any combination thereof.

Fungi could be natural, as well as mutated or genetically modified.

• Fungi further comprising an enzymatic cocktail, including one or more of cellulase, alpha-galactosidase, xylanase, glucanase, amylase, endoglucanase, hemicellulase or microbial enzyme preparations.

[0043] The fungi can be chosen to produce a desired flavor in the product since the flavor can be impacted by the fungi. In certain examples, a fungi with an acceptable flavor is Aspergillus. The product can comprise a neutral taste. In various examples, Aspergillus may provide an umami taste.

[0044] Fungi may be chosen not to produce of mycotoxins and fungi may be chosen which are GRAS (generally recognized as safe).

[0045] Different substrates contain different nutrients, therefore they can require different parameters for processing. For instance, if the major portion of carbohydrates in the substrate is a complex carbohydrate (fiber), then it will require longer time for digestion by fungi rather than simple carbohydrates (sugars). During SSF the following parameters may be used

Temperature in a range of 20-38 degrees Celsius, such as, for example, in a range of 25-36 degrees Celsius. At lower temperatures the fungi may produce proteases that destroy proteins and at higher temperatures the fungi may produce amylases that destroy complex carbohydrates. Additionally, at a temperature around 40 degrees Celsius fungi start to produce spores and stop the fermentation and the Fungi may die at temperatures greater than 40 degrees Celsius.

• The fermented ingredients from the SFF have a moisture content of 30-50% by total weight, such as, for example, 35% by total weight.

• Solid state fermentation time varies in the range of 48-96 hours, such as, for example, 60 hours to 84 hours. In various examples, the time spent in SSF is 72 hours. Less time may result in lower yields and longer times may require more bioreactors hence capital, and leads to higher costs.

• The pH of the SSF can be in a range of 1 to 14, such as, for example, 5-7

• The fermentation may occur in one or several reactors. For example, simple carbs, which typically ferment faster than complex carbs, may be fermented in a first reactor for 2 days and complex carbs can be fermented in a second reactor for a longer period than the first reactor. The first and second reactors can then be mixed upon the second extrusion.

[0046] In various other examples, different parameters may be used.

[0047] The process can be a batch and/or continuous process for improved efficiency.

Agitation may be provided to the process.

[0048] The second extrusion parameters may be:

• a moisture in the barrel: in a range of 25-80% by total weight, such as, for example, 55% by total weight. Too few or too much moisture may lead to poorer texture formation and too high moisture may also lead to pumping problems.

• a temperature in a range of 90-200C, such as, for example, 140C to 160C. In various examples, the temperature can be 150C. A temperature too low may not lead to protein fibers formation and too high may burn proteins. • a pressure in a range of 10-150 bar, such as, for example, 80-150 bar. A pressure too low may not lead to protein fibers formation and too high may result in burst.

• a speed in a range of 80-400 RPM, such as, for example 300 RPM to 400 RPM.

In various examples, the speed is 350 RPM. Speed can influence the residence time. The residence time may need to be lowered for protein fibers formation and a higher speed can increase throughput.

[0049] All the above parameters can inter-influence each other and an equilibrium may produce a desired texture.

[0050] Optionally, the second extruder can comprise a cooling die and up to 20% by total weight fat injection at the last sections of the barrel or cooling die (optional), such as, for example, 10 % to 20% by total weight. In various examples, the 15% fat is injected.

[0051] The parameters of the second extrusion may be selected in order to kill fungi present in the hydrolysate after SSF. The optimum parameters for the second extrusion to kill the fungi and produce the desired texture may depend on the raw ingredients and fungi utilized in SSF.

[0052] In the course of the research leading up to the present invention, it was unexpectedly found that the method according to present disclosure effectively works for whole crops, inherently low in protein, such as pulses and yellow pea in particular to process them into fibrous meat with similar to jerky texture as shown in on FIG 3C.

[0053] Various embodiments of the process of this invention are illustrated in the following examples wherein:

[0054] EXAMPLE #1 Oilseed meal

[0055] 100 kg of Soybean meal were extruded at a temperature 130°C, a pressure of 15 bar, and a speed in a range of 350 RPM. The extruded ingredients were subjected to solid-state fermentation in the presence of Aspergillus oryzae for 48 hours, at a moisture content of 30% by total weight of the raw ingredients and temperature of 25-30°C. The fermented ingredients were extruded at a temperature 150°C, a pressure 15 bar, and a screw speed of 350 RPM and resulted in 208 kg of fibrous meat-like product.

[0056] Table 4. Soybean meal-based meat analog

[0057] The image of the resulting product according to this process with a fibrous texture similar to red meat is shown in FIG. 2A.

[0058] EXAMPLE #2 Spent grains

[0059] Since Corn Distillers Dried Grains with Solids has small particle size and loose structure, the first extrusion step was omitted. 100 kg of Corn Distillers Dried Grains with Solids were subjected to solid-state fermentation in the presence of Aspergillus oryzae for 72 hours, at a moisture content of 30% by total weight of the raw ingredients and temperature of 25-30°C. The fermented ingredients were extruded at a temperature of 150°C, a pressure 15 bar, and a screw speed of 350 RPM and resulted in 200 kg of fibrous meat-like product.

[0060] Table 5. Com Distillers Dried Grains with Solids -based meat analog

[0061] Omitting the first extrusion step resulted in increased fermentation time and less fibrous texture. The image of the resulting meat-like product according to this process is shown in FIG. 2B.

[0062] EXAMPLE #3 Pulses

[0063] 100 kg of Yellow split pea were extruded at a temperature of 130°C, a pressure of

15 bar, and a speed in a range of 350 RPM. The extruded ingredients were subjected to solid- state fermentation in the presence of Aspergillus oryzae for 48 hours, at a moisture content of 30% by total weight of the raw ingredients and temperature of 25-30°C. The fermented ingredients were extruded at a temperature 150°C, a pressure 15 bar, and a screw speed of 350 RPM and resulted in 190 kg of fibrous meat-like product.

[0064] Table 6. Yellow pea -based meat analog

[0065] Inherently low (24.5% dry weight) protein content in Yellow pea even after more than 20% increase due to solid-state fermentation was not considered enough for extrusion into meat applications, but unexpectedly resulted in red jerky-like meat with a chewy texture.

[0066] Attributing this unexpected effect to the equilibrium of texturized protein, gelatinized starch, and Maillard reaction happened we showcase the image of the resulting product according to this process in FIG. 2C.

[0067] EXAMPLE #4. Mixes

[0068] To demonstrate the inter-influence of different parameters on of the process to different ingredients the inventor of the present application has run several processes for producing fibrous meat analogs as follows: Step 1 - Dry extrusion creates a porous structure, increasing the surface area and airflow (optional)

Step 2 - Solid-state fermentation with fungi converts carbohydrates in raw ingredients in extra protein

Step 3 - Dry extrusion de-solubilizes protein in the ingredient stream

Step 4 - Water washes away soluble carbohydrates, ash, and other soluble impurities

Step 5 - Wet extrusion works as a kill step and creates a fibrous texture similar to meat

[0069] The variations explained and key results summarized in appropriate tables below.

[0070] The extrusion at step 1 can moisten the raw ingredients in preparation for the fermentation. For example, the raw ingredients can be moistened up to 50% moisture by total weight of the raw ingredients during step 1, such as, for example, at least 10% moisture, at least 20% moisture, at least 30% moisture, or at least 40% moisture. Additionally, during step 1 and/or during step 2, nutrients (e.g., Nitrogen, minerals, vitamins) can be supplemented for better fungi growth during fermentation at step 2 and/or better nutritional value of the product for human consumption. Further, the pore size created during step 1, which in some examples can be 0.01 mm to 1 mm, can increase the surface area of the raw ingredients for growth and improve airflow during fermentation at step 2. In various examples, the extrudate exiting the extruder at step 1 has a thickness of the wall in a range of 0.7 mm to 1.5 mm.

[0071] Extrusion at step 1, step 3, and/or step 5 can comprise a temperature in a range of

90 degrees Celsius to 200 degrees Celsius, a pressure in a range of 1 bar to 150 bar, a speed in a range of 80 RPM to 400 RPM, or a combination thereof.

[0072] The Table 7 provides various example extrusion parameters that may be used for extrusion step 1.

[0073] Table 7

[0074] During fermentation at step 2, carbohydrates can be broken down and protein content can be increased. In the Tables 8, 9, 10 the raw ingredient results are measured prior to step 1 and the “HME” results are measured after solid-state fermentation and high moisture extrusion.

[0075] Table 8. Nutrients before and after solid-state fermentation and extrusion

[0076] Table 9. Sugar reduction results after solid-state fermentation and extrusion.

[0077] Table 10. Amino-acid profile before and after solid-state fermentation and extrusion

[0078] The extrusion at step 3 can help inhibit proteins from washing away during step 4 by creating protein clusters. The proteins cut by fungi enzymes into parts/amino acids during fermentation at step 2 can be washed away and the yield could be lower if extrusion was not performed at step 3. Extrusion can help to bond these parts/amino acids and keep then in the solids. Step 3 can desolubilize the protein so that the remaining carbohydrates, ash, fat, and./or non-protein substances not converted in step 2 can be removed during step 4. For example see the results for examples of protein concentrations in the following Table 11.

[0079] Table 11. Some example of extrusion parameters for extrusion step 3.

[0080] Washing at step 4 can remove carbohydrates and increase the protein concentration further, as shown in the Table 12 below

[0081] Table 12. Protein, % by dry weight after several processing steps

[0082] Extrusion at step 5 can achieve a desired texture and act as a kill step for the fermentation. In various examples, step 3 can act as a kill step for the fermentation. In various examples, the extrusion at step 5 can have the following parameters: Temperature equal to or greater than 130C, Pressure equal to or greater than 10 bar, and/or a Residence time equal to or greater than 30 sec. The extrusion parameters during steps 1, 3, and/or 5 may vary depending on the application.

[0083] The new process according to the present disclosure can produce fewer by- products (e.g., pea with its starch), utilize substrates with initially low protein content, such as fruit and vegetable peels, brans (wheat, rice), pomaces (tomato, grape, etc), further improve taste (e.g., cleaner than just fermented), and improve texture which can hold more fat and improve mouthfeel/perceived juiciness.

[0084] Summarizing the results, it is believed by inventor that applying the disclosed process may increase protein content for the substrates as shown in the table 13 below.

[0085] Table 13. Potential protein content in various ingredients after different processing steps

[0086] It is believed by inventor that applying the disclosed process may result in meat analogs with parameters as shown in the Table 14 below:

[0087] Table 14. Potential protein content in various ingredients after different processing steps

Claims

CLAIMS What is claimed is:

1. A method for a fibrous meat analog manufacturing, the method comprising: solid state fermentation of raw ingredients in the presence of fungi to convert carbohydrates into protein to form fermented ingredients; and extruding the fermented ingredients to form a fibrous meat analog structure.

2. The method of claim 1, further comprising extruding raw ingredients to create a porous structure before solid-state fermentation.

3. The method of claim 2, wherein extrudate has pores with size at least 0.01mm.

4. The method of claim 1, further comprising washing the extruded fermented ingredients with an aqueous solution to form a washed component; extruding the washed component to form a product.

5. The method of claim 1, further comprising milling the fermented ingredients before extrusion.

6. The method of claim 1, further comprising concentrating the fermented ingredients before extrusion into protein and fiber-rich fractions.

7. The method of claim 1, wherein the solid state fermentation comprising addition of an enzymatic cocktail, including one or more of cellulase, alpha-galactosidase, xylanase, glucanase, amylase, endoglucanase, hemicellulase or microbial enzyme preparations.

8. The method of claim 1, wherein the raw ingredients comprise an oilseed byproducts.

9. The method of claim 8, wherein the oilseed byproducts comprise at least one of a sunflower oilcake, a soybean oilcake, a cottonseed oilcake, a rapeseed oilcake, a canola oilcake, a copra oilcake, a palm kernel oilcake, a peanut oilcake, or a combination thereof.

10. The method of claim 1, wherein the raw ingredients comprise at least one of distillers dried grains with solubles, brewers spent grain, fruit and vegetable pomaces, bran, grains, legumes, algae or a combination thereof.

11. The method of claim 1, further comprising supplementing nitrogen, minerals, and vitamins from a mineral source, an organic source, a waste water source, or a combination thereof prior to and/or during fermentation.

12. The method of claim 1, wherein the Fungi is from the phylum Basidiomycota, Ascomycota, Glomeromycota, Mucoromycota, or Zoopagomycota.

13. The method of claim 1, wherein Fungi is from the division agaricomycotina, ustilagomycotina, pezizomycotina, saccharomycotina, taphrinomycetes, diversisporalis, archaeosporales, paraglomerales, endogonales, mucorales, mortieralles, entomophthoromycotina, asellariales, kickxellales, dimargaritales, harpellales, zoopagomycotina, or combinations thereof.

14. The method of claim 1, wherein Fungi is from the class tremellomycetes, dacrymycetes, agaricomycetes, exobasisiomycetes, ustilaginomycetes, malasseziomycetes, moniliellomycetes, arthoniomycetes, coniocybomycetes, dothideomycetes, eurotiomyctes, geoglossomycetes, laboulbeniomycetes, lecanoromycetes, leotiomycetes, lichinomycetes, orbiliomycetes, pezizomycetes, sordariomycetes, xylonomycetes, or combinations thereof.

15. The method of claim 1, wherein Fungi is from the order filobasi diales, agaricales, amylocorticiales, atheliales, boletales, jaapiales, lepidostromatales, geastrales, gomphales, hysterangiales, phallales, auriculariales, cantherellales, corticiales, gleophylalles, hymenochaetales, polyporales, russulales, sebacinales, stereopsidales, thelephorales, trechisporales, ceraceosorales, doassansiales, entyomatales, exobasidiales, georgefischeriales, microstromatales, tilletiales, urocystales, ustilaginales, malassezioales, moniliellales, saccharomycetales, coronophorales, glomeralles, hypocreales, melanosporales, microascales, boliniales, calosphaeriales, chaetospheriales, coniochaetales, diasporthales, magnaporthales, ophiostomatales, sordariales, xylariales, koralionastetales, lulworthiales, meliolales, phylachoralles, trichosphariales, eurotiales, chaetothyriales, pyrenulales, verrucariales, onygenales, mortierellales, mucorales, endogonales, or combinations thereof.

16. The method of claim 1, wherein Fungi is from the family Filobasidium, Dacromycetaceae, Agaricaceae, Amanitaceae, Bolbitiaceae, Broomeiceae, Chromocyphellaceae, Clavariaceae, Cortinariaceae, Cyphellaceae, Enolomataceae, Fistulinaceae, Himigasteraceae, Hydnangiaceae, Hygrophoraceae, Inocybaceae, Limnoperdacea, Lyophyllaceae, Marasmiaceae, Mycenacea, Niaceae, Pellorinaceae, Physalacriaceae, Pleurotacea, Pluteaceae, Porotheleaceae, Psathyrellaceae, Pterulacea, Schizophyllaceae, Stephanosporaceae, Strophariaceae, Tricholomataceae, Typhulaceae, Boletaceae, Boletinellaceae, Coniophoraceae, Diplocystaceae, Gasterellaceae, Gastrosporiaceae, Gomphidiaceae, Gyroporaceae, Hygrophoropsidaceae, Paxillaceae, Protogastraceae, Rhizopogonaceae, Sclerodermataceae, Serpulaceae, Suillaceae, Tapinellaceae, Hymenochaetaceae, Repetobasidiaceae, Schizoporaceae, Cystostereaceae, Fomitopsidaceae, Fragiporiaceae, Ganodermataceae, Gelatoporaceae, Meripilaceae,

Merulaciaea, Phenerochaetaceae, Polyporaceae, Sparassidaceae, Steccherinaceae, Xenasmataceae, Albatrellaceae, Amylostereaceae, Auriscalpaceae, Bondarzewiaceae, Echinodontiaceae, Hericiaceae, Hybogasteraceae, Lachnocladiaceae, Peniphoraceae, Russulaceae, Gloeocyctidiellacceae, Stereaceae, Ustilaginomycetes, Saccharomycetaceae, Saccharomycodaceae, Saccharomycopsidaceae, Chaetomiaceae, Lasiosphaeriaceae,

Sordariaceae, or any combination thereof.

17. The method of claim 1, wherein Fungi is from the genus Neurospora, Aspergillus, Trichoderma, Pleurotus, Ganoderma, Inonotus, Cordyceps, Ustilago, Rhizopus, Tuber, Fusarium, Pennicillium, Xylaria, Trametes, or any combination thereof.

18. The method of claim 1, wherein Fungi is from the species Aspergillus oryzae, Rhizopus oryzae, Fusarium graminareum, Cordyceps militaris, Cordyceps sinensis, Tuber melanosporum, Tuber magnatum, Pennicillium camemberti, Neurospora intermedia, Neurospora sitophila, Xylaria hypoxion, or any combination thereof.

19. The method of claim 1, wherein Fungi could be natural, as well as mutated or genetically modified.

20. The method of claim 1, wherein the extrusion parameters comprise a temperature in a range of 90-200°C, a pressure in a range of 1-150 bar, and a screw speed in a range of 80-400 RPM.

21. The method of claim 1, wherein the extrusion parameters comprise a temperature of 150°C, a pressure in a range of 15 bar, and a screw speed in a range of 350 RPM.

22. The method of claim 1, wherein the solid-state fermentation parameters comprise fermentation time of 24-72 hours, moisture content of 40-60% by total weight of the raw ingredients and temperature of 25-40°C.

23. The method of claim 1, wherein the solid-state fermentation parameters comprise fermentation time of 48 hours, moisture content of 40% by total weight of the raw ingredients and temperature of 25-30°C.

24. The method of any one of claims 22 and 23, wherein the moisture source comprises:

Water;

Wet ingredients to be mixed with extruded ingredients; or Combinations thereof.

25. The method of claim 24, wherein the wet ingredients comprise at least one of a vegetable pulp, okara, pomace slurry, brewers/distillers thin stillage, syrup, starchy drain waters.

26. The method of any one of claims 22-23, wherein the moisture source may contain extra carbon, nitrogen, mineral and vitamin containing nutrients;

27. The method of claim 1, further comprising adding secondary ingredients such as protein, carbohydrates, fat, color, flavors during or after extrusion to manufacture food product, preferably meat analog.

28. The method of claim 1, further comprising post-processing the extruded fermented ingredients, wherein the post-processing includes: mixing the extruded fermented ingredients with one or more supplemental ingredients; and/or postprocessing the extruded fermented ingredients mixed with the one or more supplemental ingredients into one or more of the following: a textured protein, patty, meatball, frankfurter, sausage, minced meat, nugget, meat analog, and a threedimensional printed food.

29. A fibrous meat analog made according to the method of claim 1.

30. A fibrous meat analog made according to the method of claim 1, wherein the meat analog comprises:

18-30% protein by dry weight of the meat analog; and 25-75% by total weight of the meat analog water.

31. An extruded meat analog from solid state fermented ingredients according to claim 30, further containing:

20-30% protein by dry weight of the meat analog; and 55-75% water

32. An extruded meat analog from solid state fermented ingredients according to claim 30, further containing:

18-30% protein by dry weight of the meat analog; and 25-35% water.

33. The product of claim 30, wherein the protein content is at least 10% higher than the protein content in raw ingredients.

34. The product of claim 30, wherein an amino acid content in the product is at least 10% higher than the amino acid content in the raw ingredient.

35. The product of claim 30, wherein a sum of total essential amino acids in the product is at least 10% higher than the sum of total essential amino acids in the raw ingredient.

36. The product of claim 30, wherein the meat analog further comprises

A hardness in a range of 4,000 N to 32,000 N; a chewiness in a range of 2 mJ to 130 mJ; a springiness in a range of 1 mm to 6 mm; a gumminess in a range of 2,000 N to 35,000 N; a chewiness of in a range of 0.5 kg to 15 kg when hydrated; a cohesiveness of 20% to 90% when hydrated; a springiness in a range of 15% to 99% when hydrated; a transversal cutting strength in a range of 3,000 to 20,000 kg/m^-2 when hydrated; a longitudinal cutting strength in a range of 2,900 to 19,000 kg/m^-2 when hydrated; average shear strength of at least 400 grams; protein fibers forming meat analog are contiguous to each other from 0.1 to 90 degree angle when viewed in a horizontal plane; said meat analog has a density of about 0.10 g/cm³ to about 0.50 g/cm³; or combinations thereof.

37. The product of claim 30, wherein the product has an average shear strength of at least 400 grams

38. A meat analog comprising:

Extruded solid state fermented ingredients;

39. An extruded meat analog from solid state fermented ingredients according to claim 38, further containing:

20-30% protein by dry weight of the meat analog; and 55-75% water

40. An extruded meat analog from solid state fermented ingredients according to claim 38, further containing:

18-30% protein by dry weight of the meat analog; and 25-35% water.

41. The product of claim 38, wherein the protein content is at least 10% higher than the protein content in raw ingredients.

42. The product of claim 38, wherein an amino acid content in the product is at least 10% higher than the amino acid content in the raw ingredient.

43. The product of claim 38, wherein a sum of total essential amino acids in the product is at least 10% higher than the sum of total essential amino acids in the raw ingredient.

44. The product of claim 38, wherein the meat analog further comprises

A hardness in a range of 4,000 N to 32,000 N; a chewiness in a range of 2 mJ to 130 mJ; a springiness in a range of 1 mm to 6 mm; a gumminess in a range of 2,000 N to 35,000 N; a chewiness of in a range of 0.5 kg to 15 kg when hydrated; a cohesiveness of 20% to 90% when hydrated; a springiness in a range of 15% to 99% when hydrated; a transversal cutting strength in a range of 3,000 to 20,000 kg/m when hydrated; a longitudinal cutting strength in a range of 2,900 to 19,000 kg/m when hydrated; average shear strength of at least 400 grams; protein fibers forming meat analog are contiguous to each other from 0.1 to 90 degree angle when viewed in a horizontal plane; said meat analog has a density of about 0.10 g/cm³ to about 0.50 g/cm³; or combinations thereof.

45. The product of claim 38, wherein the product has an average shear strength of at least 400 grams.