Fermented plant and microbial-based food products

Abstract

Described herein are non-dairy food products and methods for producing non-dairy food products using plant whole food nutrient bases and hydrocolloid fibers. The use of hydrocolloid fibers enhances the quality of non-dairy food products, such as cheese, providing a process for increased protein content compared to alternative non-dairy food products, as well as providing taste and physical properties comparable to dairy cheeses.

Description

Fermented Plant and Microbial-Based Food ProductsCROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims priority to U.S. Provisional Application No. 63 / 648,952, filed May 17, 2024, the entire contents of which are incorporated herein by reference.TECHNICAL FIELD

[0002] Described herein, are non-dairy food products and methods for producing the nondairy food products using non-dairy nutrient bases and hydrocolloid fibers. Specifically, the present disclosure provides, among other things, non-dairy cheese products and methods of making the same.BACKGROUND

[0003] The following description of the background of the present technology is provided simply as an aid in understanding the present technology and is not admitted to describe or constitute prior art to the present technology.

[0004] Non-animal food products (e.g., non-dairy cheese) have become a highly desired alternative to the animal product (e.g., dairy cheese) as they eliminate the need for animal cultivation, animal breeding programs, and animal slaughter. Many dairy cheeses, particularly natural cheeses (i.e., fermented or unprocessed cheese that contains a live flora) with an elastic and functional texture (e.g., slice-able, shred-able, etc.), can be prepared by processes that involve adding protease to a dairy milk, thus causing enzymatic coagulation of casein micelles. The protease cleaves glycomacropeptides from the casein micelles, depriving the casein micelles of a negative charge, which keeps the micelles suspended in the milk. The casein micelles collapse into a coagulated network (e.g., a curd or gel) that captures the macro elements (e.g., fats, water, and proteins) and micro elements of the milk solution into the coagulated structure.

[0005] Using non-animal ingredients to produce a similar non-animal food product, such as non-dairy cheese, has proven difficult. Indeed, currently available non-dairy and animal-freecheeses generally possess a significantly lower protein content (with most at 0% level) and lack many of the defining characteristics of certain cheeses (e.g., natural cheeses) as well as certain cheese functionalities (e.g., elasticity, chewiness, meltability, sliceability, shredability, springiness, etc.). The presently disclosed food products and methods overcome the deficiencies of currently available non-dairy and animal-free cheeses.SUMMARY

[0006] The present disclosure provides non-dairy cheeses and methods of making the same. The disclosed non-dairy cheeses are formed from a nutrient base (e.g., a nut or seed milk) and a hydrocolloid fiber. Among other benefits, the disclosed use of hydrocolloid fibers allows for full fermentation of the product, enhances the quality of non-dairy food products (e.g., non-dairy cheese) by providing a process for significantly increased protein content relative to comparable, alternative non-dairy food products, and provides textural properties like elasticity, chewiness, springiness as well as functional properties like meltability, sliceability, and shred-ability that are comparable to enzymatically coagulated dairy cheese. Further, the present disclosure provides methods that allow for eye formation and melting properties comparable to dairy cheese, as well as compositions comprising these properties.

[0007] One aspect, the present disclosure provides a food product, comprising a hydrocolloid fiber and a coagulated and fermented non-dairy nutrient base that is coagulated with the hydrocolloid fiber, wherein the food product comprises one or more eyes. In some embodiments, the one or more eyes are formed by gas-producing bacteria during fermentation of the non-dairy nutrient base.

[0008] In some embodiments, the food product comprises a non-dairy protein in an amount greater than or equal to 15% by weight based on total weight of the food product (% w / w).

[0009] In some embodiments, the non-dairy nutrient base further comprises at least one plant- or microbial -based oil. In some embodiments, the at least one plant- or microbial -based oil comprises a solid fat content (SFC) curve that is + / - 20% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 20% of a SFC curve of AMF or lard at 15°C, + / - 10% of a SFC curve of AMF or lard at 20°C, + / - 10% of a SFC curve of AMF or lard at 25°C, + / -10% of a SFC curve of AMF or lard at 30°C, + / - 10% of a SFC curve of AMF or lard at 35°C, and / or + / - 10% of a SFC curve of AMF or lard at 40°C. In some embodiments, the at least one plant- or microbial-based oil comprises a solid fat content (SFC) curve that is + / - 60% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 60% of a SFC curve of AMF or lard at 15°C, + / - 60% of a SFC curve of AMF or lard at 20°C, + / - 50% of a SFC curve of AMF or lard at 25°C, + / - 50% of a SFC curve of AMF or lard at 30°C, + / - 20% of a SFC curve of AMF or lard at 35°C, + / - 10% of a SFC curve of AMF or lard at 40°C, or any combination thereof; or, optionally, wherein the SFC curve is + / - 10% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 10% of a SFC curve of AMF or lard at 15°C, + / - 10% of a SFC curve of AMF or lard at 20°C, + / - 10% of a SFC curve of AMF or lard at 25°C, + / - 10% of a SFC curve of AMF or lard at 30°C, + / - 10% of a SFC curve of AMF or lard at 35°C, + / - 10% of a SFC curve of AMF or lard at 40°C, or any combination thereof.

[0010] In some embodiments, the at least one plant- or microbial-based oil is selected from illipe fat, coco butter, sal butter, shea butter, shea olein, shea stearin, coconut oil, coconut stearin, palm oil, palm stearin, palm olein, palm kernel, pongamia oil, and any combination thereof. In some embodiments, the at least one plant- or microbial-based oil comprises about 5% to about 70% of a stearin shea fraction and about 30% to about 95% of an olein shea fraction. In some embodiments, the plant- or microbial-based oil is present in an amount between about 10% to about 30 % w / w of the food product.

[0011] In some embodiments, the food product is a cheese.

[0012] In some embodiments, the fermented non-dairy nutrient base is a fermented plant milk or a fermented microbial-based milk. In some embodiments, the fermented plant milk comprises (i) water, and (ii) milled seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof. In some embodiments, the fermented plant milk is selected from soybean milk, lupini bean milk, almond milk, hemp seed milk, melon seed milk, pumpkin seed milk, oat milk, pea milk, fava bean milk, chickpea milk, sunflower seed milk, edamame milk, lentil milk, pistachio milk, peanut milk, walnut milk, cashew milk, coconut milk, watermelon seed milk, and macadamia milk.

[0013] In some embodiments, the hydrocolloid fiber is selected from a fiber extract from seaweed, a fiber extract from an algae, acacia gum, locust bean gum, guar gum, pectin, cellulose or a cellulose derivative, konjac, alginate, carrageenan, gellan gum, agar, pullulan, dextran, curdlan, levan, and xanthan. In some embodiments, the algae is selected from Rhodophyceae algae, Phaeophyceade algae, or Chlorophyceae algae. In some embodiments, the hydrocolloid fiber is a fiber extract from a microorganism of a genus selected from Xanthomonas, Sphingomonas, Pseudomonas, A ureobasidium, Streptococcus, Leuconostoc, Acetobacter, Azotobacter, Pseudomonas, Alcaligenes, Bacillus, Zymomonas, Aerobacter, Acetobacter, Actinomyces, Erwinia, Rhanella, Lactobacillus, Microbacterium, and Serratia.

[0014] In some embodiments, the hydrocolloid fiber forms a synergetic gel. In some embodiments, the hydrocolloid fiber is 1.0% w / w or less of the coagulated food product.

[0015] In some embodiments, the non-dairy protein is present in an amount greater than or equal to 20% w / w. In some embodiments, a protein isolate or a protein concentrate was not added to the non-dairy nutrient base or the food product.

[0016] In some embodiments, the coagulated food comprises only whole plant foods as ingredients. In some embodiments, the coagulated food product consists of 8 or fewer ingredients.

[0017] In some embodiments, the food product has a complex modulus (G*) between about 30000 PA and about 50000 PA at a frequency of 2-20 HZ. In some embodiments, the food product has a phase angle (A) between about 8 and about 16 at a frequency of 2-20 HZ. In some embodiments, wherein the food product has a cut force slope of less than 1.5 N / s.

[0018] In some embodiments, the food product is fully fermented. In some embodiments, the fermented non-dairy nutrient base is fermented with mesophilic or thermophilic bacteria. In some embodiments, the fermented non-dairy nutrient base is fermented with at least one bacteria selected from Lactococcus lactis biovar diacetylactis, Leuconostoc me senter oides, and Propionibacteria or a combination thereof.

[0019] In another aspect, the present disclosure provides method of producing a food product, comprising: (a) inoculating a non-dairy nutrient base with at least one microbialculture; (b) adding at least one hydrocolloid fiber to the non-dairy nutrient base inoculated with the at least on microbial culture, thereby inducing coagulation and formation of a curd; (c) removing water from the curd such that protein content of the curd is at least two times higher than protein content of the non-dairy nutrient base of a w / w basis; and (d) aging the curd to allow fermentation of the non-dairy nutrient base by the with at least one microbial culture comprising at least one gas-forming bacteria; wherein fermentation with the at least one gas-forming bacteria results in eye formation in the food product. In some embodiments, the at least one gas-forming bacteria is selected from Lactococcus lactis biovar diacelylaclis. Leuconostoc me senter aides, and Propionibacteria or a combination thereof.

[0020] In another aspect, the present disclosure provides a method of producing a food product, comprising: (a) homogenizing a non-dairy nutrient base; (b) inoculating the non- dairy nutrient base with at least one microbial culture; (c) adding at least one hydrocolloid fiber to the non-dairy nutrient base inoculated with the at least on microbial culture, thereby inducing coagulation and formation of a curd; (d) removing water from the curd such that protein content of the curd is at least two times higher than protein content of the non-dairy nutrient base of a w / w basis; and (e) aging the curd to allow fermentation of the non-dairy nutrient base by the with at least one microbial culture. In some embodiments, homogenizing the non-dairy nutrient base comprises high sheer homogenization, high pressure homogenization, or a combination thereof.

[0021] In some embodiments, the disclosed methods further comprise shaping the curd prior to aging the curd, wherein the shape is optionally a wheel, loaf, or a block.

[0022] In some embodiments, the disclosed methods further comprise acidification or brining of the curd after removing water and prior to aging the curd.

[0023] In some embodiments, the disclosed methods further comprise coating the curd with a water-permeable material immediately prior to aging the curd.

[0024] In some embodiments of the disclosed methods, the curd shaped in a microperforated cheese mold. In some embodiments of the disclosed methods, aging the curd is performed ata relative humidity of between about 20% to about 85% and, optionally, a temperature between about 10°C to about 22°C.

[0025] In some embodiments, the disclosed methods further comprise adding fermentation promotors or natural flavoring agents to the non-dairy nutrient base. In some embodiments, the fermentation promotors are selected from sauerkraut, miso, and nutritional yeast.

[0026] In some embodiments, the disclosed methods further comprise filtering the non-dairy nutrient base to remove solid particles prior to adding the at least one hydrocolloid fiber.

[0027] In some embodiments, the disclosed methods further comprise contacting the non- dairy nutrient base with a protease, an amino peptidase, an amylase, a cellulase, a hemicellulose, or a combination thereof prior to adding the at least one hydrocolloid fiber.

[0028] In some embodiments of the disclosed methods, the non-dairy nutrient base further comprises at least one plant- or microbial-based oil to the non-dairy nutrient base. In some embodiments, the at least one plant- or microbial-based oil comprises a solid fat content (SFC) curve that is + / - 60% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 60% of a SFC curve of AMF or lard at 15°C, + / - 60% of a SFC curve of AMF or lard at 20°C, + / - 50% of a SFC curve of AMF or lard at 25°C, + / - 50% of a SFC curve of AMF or lard at 30°C, + / - 20% of a SFC curve of AMF or lard at 35°C, + / - 10% of a SFC curve of AMF or lard at 40°C, or any combination thereof. In some embodiments, the at least one plant- or microbial-based oil is selected from illipe fat, coco butter, sal butter, shea butter, shea olein, shea stearin, coconut oil, palm oil, palm kernel, pongamia oil, and any combination thereof.

[0029] 4 In some embodiments of the disclosed methods, the at least one microbial culture comprises a thermophilic bacterial starter culture or a mesophilic bacterial starter culture.

[0030] In some embodiments of the disclosed methods, the at least one microbial culture comprises a lactic acid bacteria, a propionic acid bacteria, an acetic acid bacteria, or a yeast.

[0031] In some embodiments of the disclosed methods, the curd after removing water has a protein content of greater than or equal to 12% w / w, greater than or equal to 15% w / w, or greater than or equal to 20% w / w.

[0032] In some embodiments of the disclosed methods, the non-dairy nutrient base has a pH of 5.2 or less prior to adding the at least one hydrocolloid fiber.

[0033] In some embodiments of the disclosed methods, the non-dairy nutrient base is a plantbased milk. In some embodiments, the plant milk comprises (i) water, and (ii) milled seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof.In some embodiments, the non-dairy nutrient base comprises at least 60%, at least 65%, at least 70% at least 75%, or at least 80% w / w of water when the non-dairy nutrient base is inoculated with the at least one microbial culture.

[0034] In some embodiments of the disclosed methods, the non-dairy nutrient base is selected from soybean milk, lupini bean milk, almond milk, hemp seed milk, melon seed milk, pumpkin seed milk, oat milk, pea milk, fava bean milk, chickpea milk, sunflower seed milk, edamame milk, lentil milk, pistachio milk, peanut milk, walnut milk, cashew milk, coconut milk, watermelon seed milk, and macadamia milk.

[0035] In some embodiments of the disclosed methods, the hydrocolloid fiber forms a synergetic gel.

[0036] In some embodiments of the disclosed methods, the hydrocolloid fiber is selected from a fiber extract from seaweed, an algae, acacia gum, locust bean gum, guar gum, pectin, cellulose or a cellulose derivative, konjac, alginate, carrageenan, gellan gum, agar, pullulan, dextran, curdlan, levan, and xanthan. In some embodiments, the algae is selected from Rhodophyceae algae, Phaeophyceade algae, or Chlorophyceae algae. In some embodiments, the hydrocolloid fiber is a fiber extract from a microorganism of a genus selected from Xanthomonas, Sphingomonas, Pseudomonas, A ureobasidium, Streptococcus, Leuconostoc, Acetobacter, Azotobacter, Pseudomonas, Alcaligenes, Bacillus, Zymomonas, Aerobacter, Acetobacter, Actinomyces, Erwinia, Rhanella, Lactobacillus, Microbacterium, and Serratia.

[0037] In some embodiments of the disclosed methods, the method does not comprise adding a protein isolate or protein concentrate to the non-dairy nutrient base, the curd, or both.

[0038] In another aspect, the present disclosure provides a method of solid-state fermentation, comprising: coagulating a non-dairy nutrient base by contacting the non-dairy nutrient base with at least one hydrocolloid fiber to form a solid or gel and; inoculating the non-dairy nutrient base with at least one microbial culture comprising at least one gasforming bacteria; and incubating the solid or gel to allow fermentation of the non-dairy nutrient base by the at least one microbial culture in a solid-state, thereby producing a non- dairy cheese comprising at least one eye. In some embodiments, wherein the at least one gasforming bacteria is selected from Lactococcus lactis biovar diacelylaclis. Leuconostoc me senter aides, and Propionibacteria or a combination thereof.

[0039] In another aspect, the present disclosure provides a method of solid-state fermentation, comprising: homogenizing a non-dairy nutrient base; coagulating the non-dairy nutrient base by contacting the non-dairy nutrient base with at least one hydrocolloid fiber to form a solid or gel and; inoculating the non-dairy nutrient base with at least one microbial culture; and incubating the solid or gel to allow fermentation of the non-dairy nutrient base by the at least one microbial culture in a solid-state, thereby producing a non-dairy cheese that is capable of melting. In some embodiments, homogenizing the non-dairy nutrient base comprises high sheer homogenization, high pressure homogenization, or a combination thereof.

[0040] In some embodiments of the disclosed methods, during incubation, the non-dairy nutrient base is dehydrated at a dehydration rate consistent with a dehydration rate of dairy milk when incubating dairy cheese.

[0041] In some embodiments of the disclosed methods, during incubation, the non-dairy nutrient base is acidified at an acidification rate consistent with an acidification rate of dairy milk when incubating dairy cheese.

[0042] In some embodiments of the disclosed methods, the at least one microbial culture comprises a mesophilic bacteria or a thermophilic bacteria.

[0043] In another aspect, the present disclosure provides food products obtained by the methods disclosed herein. In some embodiments, the food product is a cheese. In some embodiments, the food product comprises only whole plant foods and the hydrocolloid fiber as ingredients. In some embodiments, the food product consists of 8 or fewer ingredients. In some embodiments, the food product has a protein content of greater than or equal to 15% w / w, or optionally greater than or equal to 20% w / w.

[0044] The foregoing general description and following detailed description are exemplary and explanatory and are intended to provide further explanation of the disclosure as claimed. Other objects, advantages, and novel features will be readily apparent to those skilled in the art from the following brief description of the drawings and detailed description of the disclosure.BRIEF DESCRIPTION OF THE DRAWINGS

[0045] FIG. 1 demonstrates how a plant or microbial-based milk may be produced.

[0046] FIG. 2 demonstrates how a plant or microbial-based milk may be processed into a plant or microbial based cheese or similar food product. Similar processing can be used to produce plant-based “meats,” plant-based “fish,” and other plant-based high-protein food products or fermented foods.

[0047] FIG. 3 shows a cross-section of a hard cheese (i.e., hard texture A).

[0048] FIG. 4 shows a cross-section of a hard cheese (i.e., hard texture B).

[0049] FIG. 5 shows a cross-section of a semi-hard cheese (i.e., semi-hard texture A).

[0050] FIG. 6 shows a cross-section of a semi-hard cheese (i.e., semi-hard texture B).

[0051] FIG. 7 shows a test procedure for the evaluation of cheese meltability.

[0052] FIG. 8 shows examples of no, slow, and regular melt cheese and a depiction of a Schreiber’s concentric circle chart used for meltability experiments.

[0053] FIG. 9 shows the melting of a reference cheese (i.e., Reference 1).

[0054] FIG. 10 shows the melting of a reference cheese (i.e., Reference 2).

[0055] FIG. 11 shows the melting of a test sample (i.e., Test IB).

[0056] FIG. 12 shows the melting of a test sample (i.e., Test 2B).

[0057] FIG. 13 shows the melting of a test sample (i.e., Test 2C).DETAILED DESCRIPTION

[0058] Dairy cheeses possess several defining characteristics. The curd or gel undergoes a high level of syneresis, which is the expulsion of excess fluid. This results in concentrating effect of the milk from which the cheese is made and can provide an approximately ten-fold or more increase in the relative amount of fat and protein in the curd compared to the original milk product. On average, dairy cheese products are about 19-22% protein compared to dairy milk, which is only approximately 2.8-3.4% protein. Dairy cheese also contains concentrated fat that is characterized by high solid fat content at room temperatures (i.e., 15-20°C), and gradual melting of solid fat crystals that results in flavor release at human body temperature. Further, dairy cheese products display a high degree of elasticity and are generally easily sliceable and shred-able. Lastly, the curd or gel holds free and chemically unbound water that allows fermentation and biodegradation to occur during aging.

[0059] Currently available non-dairy cheese products come in the form of spreads or processed blocks of plant or microbial based cheeses. Spreadable non-dairy cheeses contain a low amount of protein (e.g., <10%) and are typically not slice-able, shred-able, or crumbleable. The processed blocks mainly comprise starches and fats and generally have little or no protein content. To achieve a protein level of dairy cheeses in currently available non-dairy cheese product, protein concentrates and isolates are added, but this results in a bad taste that needs to be “masked” with added flavors and the overall protein content is still generally much lower than dairy cheese. Thus, currently available non-dairy cheese products are a poor alternative to the animal product.

[0060] Further, most non-dairy cheese that is currently on the market is not fermented, and therefore preservatives must be used to obtain stable quality and exogenous flavorants mustbe added to provide a flavor notes reminiscent of a cheese product. There are few exceptions where a liquid base is subjected to fermentation, and then processed as ingredient with starch under high heat, but even these non-dairy cheeses are not fully fermented, in that additional processes are required and ingredients are added after fermentation has ceased.

[0061] The present disclosure provides methods of using hydrocolloid fiber as a solution to this problem. The use of hydrocolloids fibers that form networks in an aqueous environment (e.g., a plant milk) allows for the formation of a curd in which a high degree of syneresis can occur and in which unbound water is held to allow for fermentation and biodegradation. The presently disclosed methods and products utilize syneresis in a way that is comparable to the process of making dairy cheese, whereas currently available non-dairy cheese have generally been produced via processes that actively avoided syneresis.

[0062] The present disclosure shows that applying fibrous extracts to non-dairy nutrient bases allows for the formation of a synergetic gel (i.e., curd), water encapsulation, and subsequent syneresis of the water. The ability of the fibrous extract to gradually expel the encapsulated water provides a previously unobtainable way to concentrate the protein found in non-dairy nutrient bases, such as nut milk, seed milk, or bean milk, and it allows to produce a high protein non-dairy product. Moreover, the product can be produced entirely by fermentation, without any further processing or ingredients required after fermentation is complete, though in some instances, additional processing or ingredient may be desired. The resulting product has exceptional texture and elasticity, and it can be easily sliced and shredded, without added protein isolates, starches, or fats. Another advantage to this technique, is the use of hydrocolloid fibers does not require high temperature treatment of the non-dairy plant or microbial base. Rather, the disclosed processes can utilize cold coagulation, which allows for inoculation (i.e., culture addition) in temperature range that is viable for most mesophilic and thermophilic starter cultures. Thus, the non-dairy food product can be produced in a natural, unprocessed way allowing for proper fermentation, preservation, and product maturation.

[0063] Further, the present disclosure builds on the data and disclosure provided inPCT / US2023 / 080490, which is herein incorporated by reference, and provides methods foreye formation in the fermented non-dairy product (e.g., non-dairy cheese) and methods for modulating the melting point of the product.Definitions

[0064] Embodiments according to the present disclosure will be described more fully hereinafter. Aspects of the disclosure may, however, be embodied in different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art. The terminology used in the description herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0065] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the present application and relevant art and should not be interpreted in an idealized or overly formal sense unless expressly so defined herein.Although not explicitly defined below, such terms should be interpreted according to their common meaning.

[0066] The terminology used in the description herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety.

[0067] The practice of the present technology will employ, unless otherwise indicated, conventional techniques of molecular biology, microbiology, chemical engineering, cell biology, and food science which are within the skill of the art.

[0068] Unless the context indicates otherwise, it is specifically intended that the various features of the disclosure described herein can be used in any combination. Moreover, the disclosure also contemplates that in some embodiments, any feature or combination offeatures set forth herein can be excluded or omitted. To illustrate, if the specification states that a complex comprises components A, B and C, it is specifically intended that any of A, B or C, or a combination thereof, can be omitted and disclaimed singularly or in any combination.

[0069] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are approximations which are varied ( + ) or ( - ) by increments of 1.0 or 0.1, as appropriate, or alternatively by a variation of + / - 15 %, or alternatively 10%, or alternatively 5%, or alternatively 2%. It is to be understood, although not always explicitly stated, that all numerical designations are preceded by the term “about”. It also is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0070] As used in the description of the invention and the appended claims, the singular forms “a,” “an” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0071] As used herein, the term “about” means the recited quantity exactly and variations within a limited range encompassing plus or minus 10% of the recited quantity. In other words, the limited range encompassed can include ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, ±1%, ±0.5%, ±0.2%, ±0.1%, ±0.05%, or smaller, as well as the recited value itself. Thus, by way of example, “about 10” should be understood to mean “10” and a range no larger than “9-11”.

[0072] As used herein, the terms “acceptable,” “effective,” or “sufficient” refer to the selection of any components, ranges, dose forms, etc. disclosed herein intend that said component, range, dose form, etc., is suitable for the disclosed purpose.

[0073] Also as used herein, “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”).

[0074] As used herein, “gas-forming bacteria” and “gas-forming bacterial strain(s)” refer to heterofermentative bacteria, which is bacteria that ferment a substrate to produce not one endproduct (homofermentative), but a number of end products, including gas, such as, for example, carbon dioxide. Thus, in some embodiments, “gas-forming bacteria” and “gasforming bacterial strain(s)” produce carbon dioxide (or another gas) in addition to another end product during fermentation. These terms can be used in context of lactic fermentation that results in an array of volatile acids, carbon dioxide, and lactic acid. In contrast, homofermentative lactic acid bacteria produce only lactic acid.

[0075] As used herein, the term “fermented” refers to a food product that has undergone an anaerobic or aerobic process by which a microbial culture (e.g., bacteria or yeast) converts carbohydrates to organic acids, gases, or alcohol and converts citrates to aromatic compounds (e.g., diacetyl and acetoin).

[0076] As used herein, the term “curd” refers to an intermediate of a food product (i.e., cheese) that has been converted from a liquid to a solid mass. A curd may be produced when milk (e.g., plant milk) or another nutrient base as disclosed herein undergoes coagulation, which can occur as a result of enzyme action, acid addition, or acid / heat addition.

[0077] As used herein, the term “hydrocolloid fiber” refers to a heterogeneous group of high molecular weight, long chain hydrophilic polymer agents that can perform gelling, maintain free and chemically unbound water, and show high degree of syneresis.

[0078] As used herein, the term “non-dairy nutrient base” refers to non-dairy milk, such as a plant milk or a microbial-based milk.

[0079] As used herein, the terms “plant milk” or “plant-based milk” may be used interchangeably to refer to a non-dairy milk alternative produced from milled plants. In general, such milks will comprise (i) water, and (ii) milled seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof. Non-limiting examples of seeds, nuts, beans, and grains that may be made into milk include from soybean, lupini bean, almond, hemp seed, melon seed (e.g., watermelon seed), pumpkin seed, oat, pea, fava bean, chickpea, sunflower seed, edamame, lentil, pistachio, peanut, walnut, cashew, coconut, and macadamia nut.

[0080] As used herein, the term “microbial-based milk” refers to a non-dairy milk alternative produced from microbes including but not limited to bacteria, yeast, and algae. For example, microbial-based milk may be produced from proteins obtained from GM & non GM microbial biomass cultivation.

[0081] As used herein, the term “plant-based oil” refers to oils derived from plant sources. There are three types of plant oil determined by the process of extracting the oil and in the nature of the resulting oil. For example, vegetable oils are extracted by placing certain parts of the plant under pressure and squeezing out the oil.

[0082] As used herein, the term “microbial-based oils” refers to oils extracted from microbes, including but not limited to bacteria, yeast, and algae.

[0083] As used herein, the term “synergetic gel” refers to a gel formed by hydrocolloid fibers capable of expelling water.

[0084] As used herein, the term “acidification” refers to inoculating a starter culture to milk (e.g., plant milk) or another nutrient base as disclosed herein to convert a sugar to an acid (e.g., lactose into lactic acid).

[0085] As used herein, the term “brining” or “salting” refers to applying salt to cheese by adding, rubbing, or soaking.

[0086] As used herein, the term “starter culture” or “fermentation starter” refers to microbiological culture which performs fermentation.

[0087] As used herein, the term “thermophilic bacteria” refers to a bacteria that thrives in high temperatures, for example, between about 35°C to about 52°C.

[0088] As used herein, the term “mesophilic bacteria” refers to a bacteria that thrives in moderate temperatures, for example, between about 15°C to about 38°C.

[0089] As used herein, the term “whole plant foods” refers to foods that have not been processed, refined or had ingredients added to them. Non-limiting examples include fruits, vegetables, legumes, nuts, seeds, or whole grains.

[0090] As used herein, the term “syneresis” refers to contraction of a gel and extraction of liquid.Coagulated Food Product

[0091] The present disclosure provides for a fermented non-dairy food product (e.g., nondairy cheese) that is coagulated with hydrocolloid fibers that are added to a nutrient base. In general, the disclosed non-dairy food product comprises a protein content higher than alternative non-dairy food products, such as currently available non-dairy cheeses, and comparable to dairy-based food products, such as dairy-based cheese. Further, the present disclosure provides for formation of eyes within the disclosed non-dairy cheeses through the fermentation process. This is a feature that is unique to the disclosed non-dairy cheese among other alternatives because most non-dairy cheeses are not produced through fermentation like conventional diary cheese.

[0092] An “eye” in a cheese is a circular hole caused by gas-producing cultures, and it is commonly associated with Swiss cheese, though Swiss cheese is not the only cheese variety that includes eyes. As disclosed herein, non-dairy cheeses can be prepared according to the disclosed methods to produce one or more eyes in the final product. Briefly, the cheese is fermented and aged with live cultures, including gas-forming bacterial species such as, e.g., Lactococcus lactis biovar diacelylaclis. Leuconostoc mesenleroides. and Propionibacteria. By following the disclosed methods, the texture properties, specifically elasticity, show close similarity to a dairy, semi-hard cheese matrix that usually develops round eyes. It has been observed that the gas produced by cheese flora does not escape the matrix but accumulates in it, therefore it is able to achieve oversaturation point & form an opening in the cheese body (i.e., an “eye”). With a proper elasticity of the matrix at the moment of eye formation, the shape of the eye becomes round.

[0093] For example, the protein content (e.g., a non-dairy protein) may be at least 12% w / w, at least 13% w / w, at least 14% w / w, at least 15% w / w, at least 16% w / w, at least 17% w / w, at least 18% w / w, at least 19% w / w, at least 20% w / w, at least 21% w / w, at least 22% w / w, at least 23% w / w, at least 24% w / w, at least 25% w / w, at least 26% w / w, at least 27% w / w, at least 28% w / w at least 29% w / w, or at least 30% w / w. In some embodiments, the proteincontent (e.g., a non-dairy protein) may be about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, about 21% w / w, about 22% w / w, about 23% w / w, about 24% w / w, about 25% w / w, about 26% w / w, about 27% w / w, about 28% w / w about 29% w / w, or about 30% w / w. In some embodiments, the protein content (e.g., a non-dairy protein) may be greater than or equal to 12% w / w, greater than or equal to 15% w / w, or greater than or equal to 20% w / w.

[0094] In some embodiment, the protein content (e.g., a non-dairy protein) may be between about 12% w / w to about 15% w / w, between about 12% w / w to about 20% w / w, between about 12% w / w to about 25% w / w, between about 15% w / w to about 20% w / w, between about 15% w / w to about 25% w / w, between about 15% w / w to about 30% w / w, between about 20% w / w to about 25% w / w, between about 20% w / w to about 30% w / w, between about 20% w / w to about 35% w / w, between about 25% w / w to about 30% w / w, or between about 25% w / w to about 35% w / w.

[0095] For the purposes of the disclosed food products (e.g., non-dairy cheese), the fermented non-dairy nutrient base from which the food product is produced may be a fermented plant milk or a fermented microbial-based milk. In general, the fermented plant milk may comprise water and one or more of milled seeds, milled nuts, milled beans, milled vegetables, or milled grain. In some embodiments, the plant milk to be fermented can be selected from soybean milk, lupini bean milk, almond milk, hemp seed milk, melon seed milk, pumpkin seed milk, oat milk, pea milk, fava bean milk, chickpea milk, sunflower seed milk, edamame milk, lentil milk, pistachio milk, peanut milk, walnut milk, cashew milk, coconut milk, watermelon seed milk, and macadamia milk.

[0096] Conventionally, non-dairy milk products were not suitable for use for cheese making as they contain low protein and high carbohydrate content. Moreover, production methods for producing prior non-dairy cheeses sought to avoid syneresis. As a result, alternative non- dairy cheese products require the addition of protein concentrates or isolates, particularly after fermentation to provide protein, and even then, protein levels of such products are well below the protein levels of dairy-food products. However, this can result in undesirable tastes. In contrast, the disclosed food products and methods do not require the addition ofexogenous protein before or after fermentation or aging. In other words, in some embodiments, a protein isolate, a protein concentrate, or a combination thereof is not added to the non-dairy nutrient base to increase protein content.

[0097] In contrast, the disclosed food products utilize hydrocolloid fiber to produce a structured matrix (e.g., a synergetic gel) after addition to a nutrient base (e.g., a plant milk or microbial milk). The structured matrix creates pockets of water that are slowly released by syneresis resulting in concentration of the protein from the original nutrient base and, ultimately, a high protein coagulated food product. In general, the hydrocolloid fiber is about 1.0% w / w or less of the coagulated food product (e.g., 1.0% w / w or less, 0.9% w / w or less. 0.8% w / w or less, 0.7% w / w or less, 0.6% w / w or less, 0.5% w / w or less, 0.4% w / w or less, 0.3% w / w or less, 0.2% w / w or less, or 0.1% w / w or less). In some embodiments, the hydrocolloid fiber may be about 0.7% w / w, about 0.65% w / w, about 0.6% w / w, about 0.55% w / w, about 0.5% w / w, about 0.45% w / w, about 0.4% w / w, about 0.35% w / w, about 0.3% w / w, about 0.25% w / w, about 0.2% w / w, about 0.15% w / w, or about 0.1% w / w of the coagulated food product.

[0098] The hydrocolloid fiber is not particularly limited so long as it is capable of forming a synergetic gel when added to a nutrient base. The hydrocolloid fiber may assist in holding chemically unbound water & be able to release this water gradually. Generally, suitable hydrocolloid fibers can be derived from plants or microbes. For example, the hydrocolloid fiber can be selected from a fiber extract from seaweed, a fiber extracted from an algae (e.g., macroalgae or microalgae), acacia gum, locust bean gum, guar gum, pectin, cellulose or cellulose derivative, konjac, alginate (e.g., sodium alginate), carrageenan (e.g., K- carrageenan), agar, gellan gum, pullulan, dextran, curdlan, levan, or xanthan. In some embodiments, the hydrocolloid fiber is a fiber from an algae, and in some embodiments, the algae may be selected from Rhodophyceae algae, Phaeophyceade algae, or Chlorophyceae algae. In some embodiments, the hydrocolloid fiber may be a fiber extract from a microorganism of a genus selected from Xanthomonas, Sphingomonas, Pseudomonas, Aureobasidium, Streptococcus, Leuconostoc, Acetobacter, Azotobacter, Pseudomonas, Alcaligenes, Bacillus, Zymomonas, Aerobacter, Acetobacter, Actinomyces, Erwinia, Rhanella, Lactobacillus, Microbacterium, and Serratia.

[0099] Dairy milk, which is generally the starting product of dairy cheese and related products, exists as an emulsion of casein micelles and butterfat droplets. The disclosed nondairy food product may comprise emulsifiers (e.g., sunflower lecithin, soy lecithin, or lecithin granules) and emulsion stabilizers to mimic the emulsion (e.g., casein micelles and butterfat droplets) and / or support dispersion of the fat globules. In some embodiments, the emulsion stabilizers can be a plant-based oil or microbial -based oil. In some embodiments, the plantbased oil can be selected from shea butter, shea olein, , palm oil, , palm olein, palm kernel oil, pongamia oil, and any combination thereof. Additionally suitable plant oils include, but are not limited to, canola oil, soy oil, olive oil, sesame oil, corn oil, flax seed oil, and rice bran oil.

[0100] In some embodiments, the plant- or microbial-based oil may comprise a solid fat content (SFC) curve that is + / - 20% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 20% of a SFC curve of AMF or lard at 15°C, + / - 10% of a SFC curve of AMF or lard at 20°C, + / - 10% of a SFC curve of AMF or lard at 25°C, + / - 10% of a SFC curve of AMF or lard at 30°C, + / - 10% of a SFC curve of AMF or lard at 35°C, and / or + / - 10% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 9% of a SFC curve of AMF or lard at 10°C, + / - 9% of a SFC curve of AMF or lard at 15°C, + / - 9% of a SFC curve of AMF or lard at 20°C, + / - 9% of a SFC curve of AMF or lard at 25°C, + / - 9% of a SFC curve of AMF or lard at 30°C, + / - 9% of a SFC curve of AMF or lard at 35°C, and / or + / - 9% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 8% of a SFC curve of AMF or lard at 10°C, + / - 8% of a SFC curve of AMF or lard at 15°C, + / - 8% of a SFC curve of AMF or lard at 20°C, + / - 8% of a SFC curve of AMF or lard at 25°C, + / - 8% of a SFC curve of AMF or lard at 30°C, + / - 8% of a SFC curve of AMF or lard at 35°C, and / or + / - 8% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 7% of a SFC curve of AMF or lard at 10°C, + / - 7% of a SFC curve of AMF or lard at 15°C, + / - 7% of a SFC curve of AMF or lard at 20°C, + / - 7% of a SFC curve of AMF or lard at 25°C, + / - 7% of a SFC curve of AMF or lard at 30°C, + / - 7% of a SFC curve of AMF or lard at 35°C, and / or + / - 7% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 6% of a SFC curve of AMF or lard at 10°C, + / - 6%of a SFC curve of AMF or lard at 15°C, + / - 6% of a SFC curve of AMF or lard at 20°C, + / - 6% of a SFC curve of AMF or lard at 25°C, + / - 6% of a SFC curve of AMF or lard at 30°C, + / - 6% of a SFC curve of AMF or lard at 35 °C, and / or + / - 6% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 5% of a SFC curve of AMF or lard at 10°C, + / - 5% of a SFC curve of AMF or lard at 15°C, + / - 5% of a SFC curve of AMF or lard at 20°C, + / - 5% of a SFC curve of AMF or lard at 25°C, + / - 5% of a SFC curve of AMF or lard at 30°C, + / - 5% of a SFC curve of AMF or lard at 35°C, and / or + / - 5% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 4% of a SFC curve of AMF or lard at 10°C, + / - 4% of a SFC curve of AMF or lard at 15°C, + / - 4% of a SFC curve of AMF or lard at 20°C, + / - 4% of a SFC curve of AMF or lard at 25°C, + / - 4% of a SFC curve of AMF or lard at 30°C, + / - 4% of a SFC curve of AMF or lard at 35°C, and / or + / - 4% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 3% of a SFC curve of AMF or lard at 10°C, + / - 3% of a SFC curve of AMF or lard at 15°C, + / - 3% of a SFC curve of AMF or lard at 20°C, + / - 3% of a SFC curve of AMF or lard at 25°C, + / - 3% of a SFC curve of AMF or lard at 30°C, + / - 3% of a SFC curve of AMF or lard at 35°C, and / or + / - 3% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 2% of a SFC curve of AMF or lard at 10°C, + / - 2% of a SFC curve of AMF or lard at 15°C, + / - 2% of a SFC curve of AMF or lard at 20°C, + / - 2% of a SFC curve of AMF or lard at 25°C, + / - 2% of a SFC curve of AMF or lard at 30°C, + / - 2% of a SFC curve of AMF or lard at 35°C, and / or + / - 2% of a SFC curve of AMF or lard at 40°C. In some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 1% of a SFC curve of AMF or lard at 10°C, + / - 1% of a SFC curve of AMF or lard at 15°C, + / - 1% of a SFC curve of AMF or lard at 20°C, + / - 1% of a SFC curve of AMF or lard at 25°C, + / - 1% of a SFC curve of AMF or lard at 30°C, + / - 1% of a SFC curve of AMF or lard at 35°C, and / or + / - 1% of a SFC curve of AMF or lard at 40°C. In other words, in some embodiments, the plant- or microbial-based oil may comprise a SFC curve that is + / - 1-10% of a SFC curve of AMF or lard at 10°C, + / - 1-10% of a SFC curve of AMF or lard at 15°C, + / - 1-10% of a SFC curve of AMF or lard at 20°C, + / - 1-10% of a SFC curve of AMF or lard at 25°C, + / - 1-10% of a SFCcurve of AMF or lard at 30°C, + / - 1-10% of a SFC curve of AMF or lard at 35°C, + / - 1-10% of a SFC curve of AMF or lard at 40°C, or any combination thereof.

[0101] The solid fat content of the disclosed non-dairy food products (e.g., non-dairy cheeses) may provide textural properties and flavor release similar or equivalent to dairy cheese. The oils or oil blends utilized in the disclosed non-dairy food products (e.g., non- dairy cheeses) can be selected to produce a melt profile comparable to anhydrous milk fat (AMF) or lard. The present inventors found that such a melt profile can be achieved, for example, using shea or palm fractions that have not undergone any modifications aside from a physical fractionation step. Thus, in some embodiments, the disclosed non-dairy food products (e.g., non-dairy cheeses) do not include hydrogenated oils, which are commonly utilized in other non-dairy alternatives that are currently marketed.

[0102] The specific blend of fats that are used in the disclosed non-dairy cheese may vary depending on the cheese and the desired attributes. In dairy cheese, the fat fraction constitutes roughly 1 / 3 of the product and plays a role on the texture, flavor, and functional properties (e.g., melting) of the cheese. Most high protein plant whole foods like seeds or nuts contain oils with low to no solid fat content, and bean lipid content is negligible However, tropical plant fats like palm, palm kernel, coconut, illipe, coco, sal and shea contain high contents of fat crystals at moderate temperatures and can be used to prepare fat blends with specific SFC (crystal melt) profiles to provide textural and sensory & melting functionality to the disclosed non-dairy food products. For example, fat blends with a olein percentage of about 40-65% (e.g., 40%, 45%, 50%, 55%, 60%, 65%) and a stearin percentage of about 35-60% (e.g., 35%, 40%, 45%, 50%, 55%, 60%) may improve viscoelastic parameters (e.g., firmness) and enhance creaminess. Fat blends with an olein percentage of about 30-40% (e.g., 30%, 35%, 40%) and a stearin percentage of about 60-70% (e.g., 60%, 65%, 70%) may reduce graininess and enhance smooth texture. And fat blends with an olein percentage of about 10-30% (e.g., 10%, 15%, 20%, 25%, 30%) and a stearin percentage of about 70-90% (e.g., 70%, 75%, 80%, 85%, 90%) may improve melting / meltability during baking or cooking. Those skilled in the art will understand how to utilize the melt curves provided in Example 11 to select a fat blend for the desired purpose.

[0103] Thus, the disclosed non-dairy food products (e.g., non-dairy cheeses) may comprise a plant- or microbial-based oil that includes about 5% to about 70% (e.g., about 20% to about 40%) of a solid stearin fraction and about 30% to about 95% (e.g., about 60% to about 80%) of liquid olein fraction. For example, the plant- or microbial -based oil may comprise about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, or about 70% of a solid stearin fraction. In some embodiments, the plant- or microbial-based oil may comprise about 20% to about 40%, about 20% to about 35%, about 20% to about 30%, about 20%, to about 25%, about 25% to about 40%, about 25% to about 35%, about 25% to about 30%, about 30% to about 40%, about 35% to about 40% of a solid stearin fraction. Similarly, the plant- or microbial-based oil may comprise about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% of a liquid olein fraction. For example, the plant- or microbialbased oil may comprise about 60% to about 75%, about 60% to about 70%, about 60% to about 65%, about 65% to about 80%, about 65% to about 75%, about 65% to about 70%, about 70% to about 80%, or about 70% to about 75% of a liquid olein fraction. Thus, the oil may comprise 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%,35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%,51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%,67%, 68%, 69%, or 70% of a solid stearin fraction, and 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%,52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%,68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, or 95% of liquid olein fraction. In some embodiments, the fractions may be shea fractions, and in some embodiments, the fractions may be palm fractions. For example, the disclosed non-dairy food products may comprise an oil blend comprising about 20% to about 40% of a shea stearin fraction and about 60% to about 80% of shea olein fraction. In some embodiments, the disclosed non-dairy food products may comprise an oil blend comprising about 25% of astearin shea fraction and about 75% of an olein shea fraction. This blend melts gradually improving product palatability and flavor release, as well as providing improved texture.

[0104] In some embodiments, the plant-based oil or the microbial-based oil is present in an amount between about 5% w / w to about 30% w / w of the coagulated food product. For example, the plant-based oil or the microbial-based oil is present in the an amount of about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, or about 30% w / w of the coagulated food product. In some embodiments, the food based oil or the microbial based oil is present in the an amount between about 5% w / w to about 10% w / w, between about 5% w / w to about 15% w / w, between about 5% w / w to about 20% w / w, between about 5% w / w to about 25% w / w, between about 10% w / w to about 15% w / w, between about 10% w / w to about 20% w / w, between about 10% w / w to about 25% w / w, between about 10% w / w to about 30% w / w, between about 15% w / w to about 20% w / w, between about 15% w / w to about 25% w / w, between about 15% w / w to about 30% w / w, between about 20% w / w to about 25% w / w, between about 20% w / w to about 30% w / w, or between about 25% w / w to about 30% w / w. In some embodiments, the plantbased oil or the microbial -based oil is present in the an amount greater than or equal to 10% w / w, greater than or equal to 12% w / w, greater than or equal to 14% w / w, greater than or equal to 16% w / w, greater than or equal to 18% w / w, greater than or equal to 20% w / w, greater than or equal to 22% w / w, greater than or equal to 24% w / w, greater than or equal to 26% w / w, greater than or equal to 28% w / w, or equal to 30% w / w.

[0105] In general, the disclosed coagulated food product (e.g., non-dairy cheese) is made from wholes plant food (and a hydrocolloid fiber, which may also be derived from a plant). In other words, the disclosed products generally do not include chemical additives. Non limiting examples of whole plant foods from which the disclosed food products may be derived include, but are not limited to, nuts, seeds, grains, beans, fruits, and vegetables. In some embodiments, the coagulated food product only comprises (i.e., consists of or consists essentially of) whole plant foods, in addition to the hydrocolloid fiber. In some embodiments, the coagulated food product consists of 10 or fewer ingredients. In some embodiments, thecoagulated food product consists of 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, or 9 or more ingredients. For example, a coagulated food product (e.g., non-dairy cheese) may be made from a non-dairy nutrient base comprising about 20 to about 40% seeds, beans, or nuts, about 1% to about 5% of sauerkraut, about 1% to about 10% of miso, about 1% to about 2% of nutritional yeast, and about 43% to about 77% water.

[0106] The disclosed non-dairy cheeses are unique among non-dairy cheese alternatives because the disclosed non-dairy cheeses possess functional characteristics of dairy cheese (e.g., slice-ability, shred-ability, chewiness, meltiness, etc.) that lack in currently available dairy cheese alternatives that were on the market prior to this application. As shown in Example 7, the disclosed non-dairy cheeses possess elasticity, stiffness, and chewiness that are comparable to dairy cheese, whereas no other currently marketed non-dairy cheese alternative comes close.

[0107] In particular, the disclosed non-dairy cheeses may possess a complex modulus (G*; a measure of elasticity / stiffness) comparable to gouda cheese, i.e., a complex modulus of about 30000-50000 PA when measured at a frequency between 20-20 HZ. Similarly, the disclosed non-dairy cheeses may possess a phase angle (5; a measure of solid / liquid balance) comparable to gouda cheese, i.e., about 8-16 when measured at a frequency between 20-20 HZ. Finally, whereas most currently marketed non-dairy cheese alternatives are not suitable for slicing or shredding or are more difficult to slice and shred, the disclosed non-dairy cheeses slice and shred similar to diary cheese, such as gouda. For example, the disclosed non-dairy cheese may have a cut force slope of less than 1.5 N / s or between 1-1.5 N / s. For the purposes of the present disclosure, any of the desirable functional features of cheese can be measured according to the method provided in Example 7.Methods for Producing a Fermented / Coagulated Non-Dairy Food Product

[0108] The disclosed methods are the first to use a solid-state fermentation approach to creating a non-dairy fermented food product, such as a non-dairy cheese. These methods mirror the process of making dairy cheese and therefore yield a similar, high-protein product that behaves much like dairy cheese. The disclosed methods allow for acidification anddehydration that corresponds to the acidification and dehydration that occur during the process of making dairy cheese, thus underscoring the unexpected ability of the disclosed methods to provide a non-dairy food product (e.g., a non-dairy cheese) that is exceptionally similar to dairy cheeses.

[0109] Thus, another aspect of the disclosure is directed to a method for producing a non- dairy food product (e.g., a non-dairy cheese) that generally has higher protein content (e.g., an amount of a non-dairy protein) than alternative non-dairy food products. In some embodiments, the method comprises (a) inoculating a non-dairy nutrient bases as disclosed herein, with at least one microbial culture, (b) adding at least one hydrocolloid fiber as disclosed herein to the non-dairy nutrient based inoculated with the at least one microbial culture, thereby inducing coagulation and formation of a curd; (c) removing water from the curd such that the protein content of curd is at least two times higher than the protein content of the non-dairy nutrient base.

[0110] Additionally, the present disclosure also provides methods for producing a non-dairy cheese with melting properties that are similar to conventional dairy cheeses. Without being bound by theory, the melting properties of a non-dairy cheese can be improved when the protein structure of the non-diary nutrient base is opened, thus allowing for better incorporation of the protein in oil-in-water emulsion. Selecting a fat blend with an appropriate SFC curve (e.g., a fat blend with a SFC curve similar to fat blends 6 to 9 prepared in Example 2) can also further improve the melting properties of the cheese. For, examples, a fat blend with a SFC curve that improves melting properties may comprise an olein fraction and a stearin fraction, wherein the olein fraction makes up between about 50% and about 20% (e.g., about 50%, about 45%, about 40%, about 35%, about 30%, about 25%, or about 20%) of the oil blend and the stearin fraction makes up between about 40% and about 80% (e.g., about 80%, about 75%, about 70%, about 65%, about 60%, about 55%, about 50%, about 45%, or about 40%) of the oil blend.[OHl] In order to open the protein structure of the non-dairy nutrient base, the non-dairy nutrient base can be homogenized prior to coagulation and fermentation. The homogenizationmay be high sheer homogenization, high pressure homogenization, or a combination thereof.In some embodiments, high pressure homogenization may be preferred.

[0112] Microbial culture can be selected from a thermophilic bacterial starter culture or a mesophilic bacterial starter culture. In some embodiment, the microbial culture is selected from lactic acid bacterium, a propionic acid bacterium, an acetic acid bacterium, or a yeast. In some embodiments, microbial culture is selected from the lactic acid bacteria genus of Lactobacillus, Leuconostoc, Pediococcus, Lactococcus, Steptococcus, Aerococcus, Corynebacterium, Oenococcus, Sporolactobacillus, Teragenococcus, Vagococcus, or Weissella, as well as acetic acid bacteria like Acetobacter and Gluconobacter and yeasts like Kluyveromyces, Pichia, and Saccharomyces. In some embodiments, the microbial culture is selected from the Lactococcus species. In some embodiments the Lactococcus species is Lactococcuss lactis. In some embodiments, the microbial culture is selected from Lactococcuss lactis subsp. cremoris, Lactococcuss lactis subsp. hordniae, Lactococcuss lactis subsp. lactis, Lactococcuss lactis subsp. tructae, o Lactococcuss lactis subsp. lactis biovar diacetylactis. In some embodiments, the microbial culture is selected from the Leuconostoc species. In some embodiments, the Leuconostoc species is Leuconostoc lactis or Leuconostoc me senter oides. In some embodiments, the microbial culture is selected from the Streptococcus species. In some embodiments, the Streptococcus species is Streptococcus thermophilus. In some embodiments, the microbial culture is selected from Lactobacillus species. In some embodiments, the Lactobacillus species is selected from Lactobacillus bulgaricus, Lactobacillus lactis, or Lactobacillus helveticus. In some embodiments, the microbial culture is selected from the propionic acid bacteria genus of Propionibacterium. In some embodiments, the microbial culture is selected from the Propinibacterium species. In some embodiments, the Propinibacterium species is Propionibacterium freudenreichii . In some embodiments, the microbial culture is selected from the Propionibacterium freudenreichii subsp. shermanii. In some embodiments yeast culture is a Kluyveromyces lactis culture. In some embodiments, the microbial culture is a vegan culture. In some embodiments, the culture can be selected from cultures VEGE 022, VEGE 033, VEGE011, VEGE061, LB3, VEGETAL, CHN19, Flora Danica, L100, L200, L700, X400, C08, C21, and P100.

[0113] For the purposes of preparing a non-dairy cheese comprising one or more eyes, the culture that is used for fermentation should comprise at least one gas-forming bacterial strain, such as at least one bacteria selected from Lactococcus lactis biovar diacelylaclis.Leuconostoc me senter aides, and Propionibacteria or a combination thereof.

[0114] As disclosed herein, hydrocolloid fiber is applied to the non-dairy nutrient base to generate a synergetic gel which generally induces coagulation and formation of curd. This structured matrix creates pockets of water, and the water is slowly removed from the curd by syneresis. This process results in the protein content being at least two times higher than the protein content of the non-dairy nutrient base. In some embodiments, the protein content is at least three times higher, at least four times higher, at least five times higher, at least 6 times higher, at least 7 times higher, or at least 8 times or more higher than the protein content of the non-dairy nutrient bases.

[0115] In some embodiments, the curd after removing water has a protein content (e.g., the amount of a non-dairy protein) greater than or equal to 12% w / w, greater than or equal to 15% w / w, or great than or equal to 20% w / w. For example, the protein content may be about 12% w / w, about 13% w / w, about 14% w / w, about 15% w / w, about 16% w / w, about 17% w / w, about 18% w / w, about 19% w / w, about 20% w / w, about 21% w / w, about 22% w / w, about 23% w / w, about 24% w / w, or about 25% w / w.

[0116] The disclosed curd is aged to allow for fermentation, water loss and concentration of the constituents of the nutrient base, and flavor development, which may occur through microbial and / or enzymatic degradation of the constituents of the nutrient base. In some embodiments, the curd is aged for least then one day. In some embodiments, the curd is aged for less than one day. In some embodiments, the curd is aged between about 1 day to about 10 days. In some embodiments, the curd is aged for between about 1 day to about 2 days, between about 1 day to about 3 days, between about 1 day to about 4 days, between about 2 days to about 3 days, between about 2 days to about 4 days, between about 2 days to about 5 days, between about 3 days to about 4 days, between about 3 days to about 5 days, between about 3 days to about 6 days, between about 4 days to about 5 days, between about 4 days to about 6 days, between about 4 days to about 7 days, between about 5 days to about 6 days,between about 5 days to about 7 days, or between about 6 days to about 7 days. In some embodiments, the curd is aged for at least Iweek to about 2 months. In some embodiments, the curd is aged for between about 1 week to about 2 weeks, between about 1 week to about 2 weeks, between about 1 week to about 3 weeks, between about 2 weeks to about 3 weeks, between about 2 weeks to about 4 weeks, or between about 3 weeks to about 4 weeks. In some embodiments, the curd is aged for between about 1 month to about 1.5 months, between about 1 month to about 2 months, or between about 1.5 months to about 2 months. In some embodiments, the curd is aged for between about 2 months to about 2 years. In some embodiments, the curd is aged for about 2 months to about 4 months, between about 2 months to about 6 months, between about 2 months to about 8 months, between about 4 month to about 6 months, between about 4 months to about 8 months, 4 months to about 10 months, between about 6 months to about 8 months, between about 6 months to about 10 months, between about 6 months to about 12 months, between about 8 months to about 10 months, between about 8 months to about 12 months, between about 8 months to about 14 months, between about 10 months to about 12 months, between about 10 months to about 14 months, between about 10 months to about 16 months, between about 12 months to about 14 months, between about 12 months to about 16 months, between about 12 months to about 18 months, between about 14 months to about 16 months, between about 14 months to about 18 months, between about 14 months to about 20 months, between about 16 months to about 18 months, between about 16 months to about 20 months, between about 16 months to about 22 months, between about 18 months to about 20 months, between about 18 months to about 22 months, between about 18 months to about 24 months, between about 20 months to about 22 months, between about 20 months to about 24 months, or between about 22 months to about 24 months. In some embodiments, the aging is performed in humid conditions. In some embodiments, the relative humidity is between about 20% to about 85%. In some embodiments, the relative humidity is between about 20% to about 25%, between about 20% to about 30%, between about 20% to about 35%, between about 25% to about 30%, between about 25% to about 35%, between about 25% to about 40%, between about 30% to about 35%, between about 30% to about 40%, between about 30% to about 45%, between about 35% to about 40%, between about 35% to about 45%, between about 35% to about 50%, between about 40% to about 45%, between about 40% to about 50%, between about 40% toabout 55%, between about 45% to about 50%, between about 45% to about 55%, between about 45% to about 60%, between about 50% to about 55%, between about 50% to about 60%, between about 50% to about 65%, between about 55% to about 60%, between about 55% to about 65%, between about 55% to about 70%, between about 60% to about 65%, between about 60% to about 70%, between about 60% to about 75%, between about 65% to about 70%, between about 65% to about 75%, between about 65% to about 80%, between about 70% to about 75%, between about 70% to about 80%, between about 70% to about 85%, between about 75% to about 80%, between about 75% w / w to about 85%, or between about 80% to about 85% w / w.

[0117] Microperforated molds stimulate the process of syneresis releasing the water through the micro-perforations outside of the mold. This strengthens the structure and delivers a wheel, loaf, or block of non-dairy cheese than can be handled manually. Microperforated molds play a role in the enhanced protein concentration process. In some embodiments, the coagulation process is performed in a microperforated cheese mold. In some embodiments, the block, loaf, or wheel formation process is performed in a microperforated cheese mold. In some embodiments, the acidification process is performed in a microperforated cheese mold. In some embodiments, cheese is shaped or molded in a microperforated cheese mold.

[0118] In some embodiments, the curd is shaped after the water is removed and prior to aging. In some embodiments, the curd is shaped in a wheel shape or a block shape.

[0119] Brining or salting of the curd is generally used to add flavor and to act as a preservative so the non-dairy food product (e.g., non-dairy cheese) does not spoil during the aging process. These processes can be performed by adding salt directly to the curd, the outside of the non-dairy cheese product can be rubbed with salt or with a damp cloth that has been soaked in brine (e.g., heavily salted water), or the cheese can be bathed directly in a vat of brine. In some embodiments, the method for producing a non-dairy food product further comprises acidification or brining of the curd after removal of water and prior to aging the curd.

[0120] In some embodiments, the method for producing a non-dairy food product further comprises coating the curd with a water-permeable material (e.g., polymer mix) immediately prior to aging.

[0121] The fermentation process generally allows for natural development of flavor via fermentation from the ingredients that were used to produce the non-dairy food product. During fermentation, live microflora are formed. In some embodiments, fermentation promoters or natural flavoring agents are added to the non-dairy nutrient base. In some embodiments, the fermentation promoters are selected from sauerkraut, miso, or nutritional yeast.

[0122] The present disclosure shows that fermentation using the disclosed methods can be carried out using mesophilic or thermophilic bacteria, and the resulting acidification profiles are virtually the same as dairy milk. Fermentation promoters, such as miso, sauerkraut, dextrose, and saccharolytic enzymes (e.g., hemicellulase and a-amylase) can be used to tune the acidification profile and / or correspond to a diary cheese acidification profile as shown in Example 7. Further, altering the amount of dry matter (DM) can be used to optimize the acidification profile as well. Those skilled in the art will understand how to adjust such parameters based on the Examples provided herein (e.g., Example 7).

[0123] In some embodiments, the method for producing a non-dairy food product further comprises filtering the non-dairy nutrient base (e.g., the plant milk or microbial-based milk that comprises water and either mill seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof) to remove solid particles. In some embodiments, the filtering is performed before adding the hydrocolloid fiber.

[0124] In some embodiments, the non-dairy nutrient base is contacted with a protease, an amino peptidase, an amylase, a cellulase, a hemicellulose, or a combination thereof. In some embodiments, contact occurs prior to the adding the hydrocolloid fiber.

[0125] In some embodiments, the method for producing a non-dairy food product further comprises adding at least one plant-based oil or at least one microbial-based oil. In some embodiments, the plant-based oil can be selected from shea butter, shea olein, shea stearin,illipe fat, coco butter, sal butter, coconut oil, palm oil, palm kernel, pongamia oil, or any combination thereof. In some embodiments, the plant-based oil or the microbial based oil may comprise a solid fat content (SFC) curve that is + / - 20% (e.g., + / - 5%, 10%, 15%, or 20%) of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 20% (e.g., + / - 5%, 10%, 15%, or 20%) of a SFC curve of AMF or lard at 15°C, + / - 10% (e.g., + / - 5% or 10%) of a SFC curve of AMF or lard at 20°C, + / - 10% (e.g., + / - 5% or 10%) of a SFC curve of AMF or lard at 25°C, + / - 10% (e.g., + / - 5% or 10%) of a SFC curve of AMF or lard at 30°C, + / - 10% (e.g., + / - 5% or 10%) of a SFC curve of AMF or lard at 35°C, and / or + / - 10% (e.g., + / - 5% or 10%) of a SFC curve of AMF or lard at 40°C.

[0126] In some embodiments, the non-dairy nutrient base has a pH of 5.2 or less prior to adding the hydrocolloid fiber. In some embodiments, the pH of the non-dairy nutrient base prior to adding the hydrocolloid fiber is a pH of 5 or less, 4.5 or less, 4 or less, a pH of 3.5 or less, a pH of 3 or less, a pH of 2.5 or less, a pH of 2 or less, a pH of 1.5 or less or a pH of 1 or less.

[0127] In some embodiments, the method for producing a non-dairy food product comprises using a non-dairy nutrient base that is a plant milk, and the plant milk comprises water and either milled seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof.

[0128] In some embodiments, the non-dairy nutrient base comprises at least 60% w / w of water, at least 65% w / w of water, at least 70% w / w of water, at least 75% w / w of water, or at least 80% w / w of water. In some embodiments the non-dairy nutrient base comprises between about 60% w / w to about 65% w / w of water, between about 60% w / w to about 70% w / w of water, between about 60% w / w to about 75% w / w of water, between about 65% w / w to about 70% w / w of water, between about 65% w / w to about 75% w / w of water, between about 65% w / w to about 80% w / w of water, between about 70% w / w to about 75% w / w of water, between about 70% w / w to about 80% w / w of water, or between about 75% w / w to about 80% w / w of water.

[0129] In some embodiments, the non-dairy nutrient base is selected from soybean milk, lupini bean milk, almond milk, hemp seed milk, melon seed milk, pumpkin seed milk, oatmilk, pea milk, fava bean milk, chickpea milk, sunflower seed milk, edamame milk, lentil milk, pistachio milk, peanut milk, walnut milk, cashew milk, coconut milk, watermelon seed milk and macadamia milk.

[0130] In some embodiments, the hydrocolloid fiber is added to form a synergetic gel, wherein the hydrocolloid fiber, is a fiber extract. In some embodiments, the fiber extract is from seaweed, a fiber extracted from algae (e.g., macro algae or micro algae), acacia gum, locust bean gum, guar gum, pectin, cellulose or cellulose derivative, konjac, alginate (e.g., sodium alginate), carrageenan (e.g., K-carrageenan), gellan gum, agar, pullulan, dextran, curdlan, levan, or xanthan. In some embodiments the hydrocolloid fiber is a fiber from an algae and the algae may be selected from Rhodophyceae algae, Phaeophyceade algae, or Chlorophyceae algae. In some embodiments, the hydrocolloid fiber may be a fiber extract from a microorganism of a genus selected from Xanthomonas, Sphingomonas, Pseudomonas, Aureobasidium, Streptococcus, Leuconostoc, Acetobacter, Azotobacter, Pseudomonas, Alcaligenes, Bacillus, Zymomonas, Aerobacter, Acetobacter, Actinomyces, Erwinia, Rhanella, Lactobacillus, Microbacterium, and Serratia. In some embodiments, for producing a non-dairy food product does not comprise adding a protein isolate or protein concentrate to the non-dairy nutrient base (e.g., the plant-based milk or microbial-based milk), the curd, or both.

[0131] Dairy cheese is a product of fermentation that is difficult to reproduce because a solid matrix needs to be subjected to fermentation, not a liquid. The system must be designed in a way that while matrix remains solid, a sufficient amount of chemically unbound water is present for the culture to grow, dominate the environment, convert present sugars into acid at a fast rate, thus preventing growth of contaminants and preserving the food as a consequence.

[0132] The present disclosure provides novel methods of solid-state fermentation, comprising: coagulating a non-dairy nutrient base by contacting the non-dairy nutrient base with at least one hydrocolloid fiber to form a solid or gel; inoculating the non-dairy nutrient base with at least one microbial culture; and incubating the solid or gel to allow fermentation of the non-dairy nutrient base by the with at least one microbial culture in a solid-state. The disclosed methods of solid-state fermentation are distinguishable from prior methods used toprepare non-dairy foods, and specifically non-dairy cheeses. Most non-dairy cheeses that are currently on the market are not fermented at all, though a few exceptions utilize a liquid base subjected to fermentation and then processed with starch under high heat. The solid-state fermentation methods disclosed herein provide for a novel approach for preparing a fully fermented product that does not require further processing or ingredients after fermentation has completed, and which generally allows for maintaining a live culture in the product. Optionally, a fermented product (e.g., a non-dairy cheese) created by the disclosed methods may be further processed or used as an ingredient in a further food product, but such further processing in not necessary to obtain an edible, fermented food.

[0133] Utilizing the disclosed processes and methods it is possible to prepare a non-dairy cheese product with comparable taste, texture, elasticity, and protein content to dairy cheeses. Table 1 details examples of steps that can be used to produce a non-dairy food product (e.g. non-dairy cheese) as described herein compared to steps generally used to produce dairy cheese and further described in FIG. 2. An example of how a plant-based or microbial based milk is produced is shown in FIG. 1.Table 1 - A comparison between plant based or microbial based cheese and dairy cheese production

[0134] The gel or curd in step 4 of plant based or microbial based cheese is produced by hydrocolloid fiber and the gel or curd of step 4 in dairy cheese is produced by casein. In steps 5-8 of both processes, there is a general increase in solid content as the cheese ripens and matures and the protein content of the dairy milk or non-dairy milk is approximately 3% and the resulting protein content is generally approximately 20%.

[0135] A food product can be produced by the methods as disclosed herein. Non-limiting examples include non-dairy food products such as cheese. The disclosed food product has higher protein levels than other alternative non-dairy food products (e.g., alternative nondairy cheeses). In some embodiments, the food product comprises only whole plant foods and a hydrocolloid fiber as ingredients. In some embodiments, the food product consists of 8 or less ingredients. In some embodiments, the food product has a protein content (e.g., an amount o f a non-dairy protein) of greater than or equal to 12% w / w, greater than or equal to 14% w / w, greater than or equal to 15% w / w, greater than or equal to 16% w / w, greater than or equal to 17% w / w, or greater than or equal to 18% w / w.

[0136] Generally, a food product as disclosed herein (e.g., non-dairy cheese) is sliceable, shred-able, and crumble-able as compared to alternative non-dairy cheeses. The disclosed food product is elastic, meltable, and has enhanced sensory evaluation (e.g., taste and smell) relative to other, non-dairy cheese products. In particular, the disclosed food products possess elasticity, chewiness, meltability, sliceability, shred-ability, and springiness that is comparable to enzymatically-produced dairy cheese. In contrast, few if any other currently available non-dairy cheeses are sliceable or shred-able, and other such products lack elasticity, meltability, and springiness that is similar to dairy cheese. Instead, other such products, such as starch-based non-dairy cheeses are brittle, which is both undesirable when being consumed and results in significant loss of the product during production (e.g., during slicing or shredding) and transportation. Thus, the disclosed food products possess physically distinct and desirable characteristics that are believed to be obtained as a result of the combination or ingredients and preparation process disclosed herein. For example, without being bound by theory, it is believed that the desirable texture, melting profile, and gradual flavor release of the disclosed non-dairy cheeses may be related, at least in part, to the solid fat content of the disclosed cheeses.

[0137] The present technology is not to be limited in terms of the particular embodiments described in this application, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and products within the scope of the present technology,in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the present technology. It is to be understood that this present technology is not limited to particular methods, reagents, compounds compositions or biological systems, which can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0138] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0139] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member. Thus, for example, a group having 1% w / w-3% w / w refers to groups having 1% w / w, 2% w / w, or 3% w / w. Similarly, a group having 1% w / w-5% w / w refers to groups having 10% w / w, 20% w / w, 3% w / w, 4% w / w, or 5% w / w, and so forth.

[0140] Other aspects are set forth within the following claims.EXAMPLES

[0141] The present technology is further illustrated by the following Examples, which should not be construed as limiting in any way. The examples herein are provided to illustrate advantages of the present technology and to further assist a person of ordinary skill in the art with preparing or using the compositions and systems of the present technology. Theexamples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims. The examples can include or incorporate any of the variations, aspects, or embodiments of the present technology described above. The variations, aspects, or embodiments described above may also further each include or incorporate the variations of any or all other variations, aspects or embodiments of the present technology. The following Examples build on the data provided in PCT / US2023 / 080490, and provide methods for eye formation in the fermented non-dairy product (e.g., non-dairy cheese) and methods for modulating the melting point of the product.EXAMPLE 1: Eye formation and open-structure cheese

[0142] The purpose of this experiment was to determine ripening conditions that would result in the formation of round eyes in a non-dairy cheese product. The experimental design included preparation of a matrix with 2 different levels of the texture hardness and activation of gas forming flora at different time points during cheese ripening.

[0143] Hardness variation was achieved by replacement of native seed oil with two different fat blends shown in the Table below.Table 2 - Fat blend fractions

[0144] Fractions of shea olein (Bassao E) and shea stearin (Illexao) were obtained from oils and fats manufacturer, AAK. Both fractions were fully melted with a heat blanket, combined in the above ratios, and subjected to fast crystallization in the freezer.

[0145] As gas forming culture, a “Propionib acterium 50” culture was used from Biena Inc. The activation of the culture to produce gas was done by increasing the ripening temperature of the cheese to 20°C at different stages.Table 3 - Experimental Design

[0146] The following milk was prepared for all cheeses:Table 4 - Milk base composition

[0147] Further milk processing is described in PCT / US2023 / 080490. Briefly, After hydration the whole mix was transferred to a blender (Vitamix, Explorian 310) and milled for a total time of 8 min at speed level 6. The milled base was filtered manually using an NMO filter bag of 100 pm pore size.

[0148] For further incorporation of fat, the native seed oil was removed by centrifugation for 10 min at 4700rpms in a Sorvall ST40 centrifuge manufactured by Thermo Scientific.

[0149] Skimmed milk was then transferred to a Thermomix TM6, where agar-agar powder was added during agitation in the amount of 0.52%. The milk was then pasteurized to 85°C for 5 min and cooled to the inoculation temperature of 43°C. Once the temperature was reached, a vegan culture was added (Vege 022, DuPont) in the amount of 0.02%. At this stage, the milk had a protein content ranging between 6.56% and 7.32% of protein. After culture addition, milk was transferred to a microperforated mold (Laude, 0150, SI 10040C) and the mold was kept at 20°C for the consequent 16 hours to allow coagulation and acidification process to take place. Monitoring of the acidification was accomplished with a pH data logger (Hanna, Edge) by inserting the probe in the center of the cheese wheel. The coagulated wheels of cheese were removed from the molds, weighed, and brined for 2 hours in 18% NaCl brine at 14°C. The brined cheese was then coated with PVA coating and placed in an incubator with RH<80% and a temperature of 13°C. Over a period of 6 weeks the cheese was flipped periodically, and a new layer of coating was applied once a week. Cheese wheels were dehydrated this way to desired texture, after which samples of the products were collected for compositional analyses.

[0150] For the “Hard texture cheese” a Blend 7 was used & for the “Semi-hard texture cheese” a Blend 4 was used. Fat blends were incorporated into milk with the use of high shear mixer IKA Magic Lab at speed of 21000rpms, temperature of 55°C during 10 min.

[0151] Cheese was aged for a total period of 6 weeks, cooled down & cut. Eyes were measured & counted on the cross section. FIGs. 3-6 illustrate the results.

[0152] While both semi-hard cheeses had some eyes, the semi-hard cheese texture B (FIG. 6) was most pronounced. The cheese developed multiple round eyes with an average diameter of 6mm & no cracks or slits. Semi-hard texture cheese A developed less eyes and the eyes that were formed started cracking (FIG. 5). Experiments with Hard texture cheese A & Bwere successful in terms of over saturation & texture opening, but they were not successful in terms of formation of round eyes (FIGs. 3 & 4).EXAMPLE 2: Melting Properties

[0153] Without being bound by theory, the inventors believed that melting properties can be enabled in developed cheese matrix when the structure of the “storage plant protein” will be opened & properly incorporated in the oil-in-water emulsion. Moreover, inventors believed that the fat blend sfc curve will play a significant role in the melting behavior of the cheese.

[0154] The protein structure was opened by 2 means: (a) Mechanically - by running milk through high sheer homogenizer (IKA Colloidal Mill) & high pressure homogenizer (GEA Niro Soavi Ariete); or (b) Biologically - by inoculating milk with highly proteolytic culture (L100, DSM) & subjecting the cheese to 6 week aging period.

[0155] The preparation of the cheese was otherwise carried out essentially is described in Example 1, with fat blends were incorporated into milk with the use of high shear mixer IKA Magic Lab at speed of 21000rpms, temperature of 55°C during 10 min.

[0156] The experimental design followed is described in the table below:Table 5 - Experimental Setup

[0157] High sheer homogenization was carried out with an IKA Colloidal mill at 21000rpms, 55°C during 10 min. High pressure homogenization was carried out with a GEA homogenizer Niro Soavi Ariete at 55°C during 2 step-homogenization at 210 & 50Ba.

[0158] Results validation was carried out according to the protocol in FIG. 7, with baking temperature increased to 180°C for 15 min and 220°C for 10 min.

[0159] The results summary is presented in the table below:Table 6 - Experimental Results

[0160] Results: Test 2 -B achieved a restricted melt of 2 on Schreiber scale, but Test 2 - C achieved a melt comparable with dairy young Gouda cheese with no difference at 180°C / 15 min and 3 -point difference (Schreiber 5 vs 8) at 220°C / 10 min. It was concluded that achievement of melting properties comparable with dairy cheese is possible by using an appropriate fat blend and high pressure homogenization or a similar mechanism for opening the protein structure of the non-dairy base that is used.

Claims

WHAT IS CLAIMED IS:

1. A food product, comprising a hydrocolloid fiber and a coagulated and fermented nondairy nutrient base that is coagulated with the hydrocolloid fiber, wherein the food product comprises one or more eyes.

2. The food product of claim 1, wherein the one or more eyes are formed by gasproducing bacteria during fermentation of the non-dairy nutrient base.

3. The food product of claim 1 or 2, wherein the food product comprises a non-dairy protein in an amount greater than or equal to 15% by weight based on total weight of the food product (% w / w).

4. The food product of any one of claims 1-3, wherein the non-dairy nutrient base further comprises at least one plant- or microbial-based oil.

5. The food product of claim 4, wherein the at least one plant- or microbial -based oil comprises a solid fat content (SFC) curve that is + / - 20% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 20% of a SFC curve of AMF or lard at 15°C, + / - 10% of a SFC curve of AMF or lard at 20°C, + / - 10% of a SFC curve of AMF or lard at 25°C, + / - 10% of a SFC curve of AMF or lard at 30°C, + / - 10% of a SFC curve of AMF or lard at 35°C, and / or + / - 10% of a SFC curve of AMF or lard at 40°C.

6. The food product of claim 4, wherein the at least one plant- or microbial-based oil comprises a solid fat content (SFC) curve that is + / - 60% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 60% of a SFC curve of AMF or lard at 15°C, + / - 60% of a SFC curve of AMF or lard at 20°C, + / - 50% of a SFC curve of AMF or lard at 25°C, + / - 50% of a SFC curve of AMF or lard at 30°C, + / - 20% of a SFC curve of AMF or lard at 35°C, + / - 10% of a SFC curve of AMF or lard at 40°C, or any combination thereof; or, optionally, wherein the SFC curve is + / - 10% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 10% of a SFC curve of AMF or lard at 15°C, + / - 10% of a SFC curve of AMF or lard at 20°C, + / - 10% of a SFC curve of AMF or lard at 25°C, + / - 10% of a SFC curve of AMF or lard at 30°C, + / - 10% of aSFC curve of AMF or lard at 35°C, + / - 10% of a SFC curve of AMF or lard at 40°C, or any combination thereof.

7. The food product of any one of claims 4-6, wherein the at least one plant- or microbial-based oil is selected from illipe fat, coco butter, sal butter, shea butter, shea olein, shea stearin, coconut oil, coconut stearin, palm oil, palm stearin, palm olein, palm kernel, pongamia oil, and any combination thereof.

8. The food product of any one of claims 4-7, wherein the at least one plant- or microbial-based oil comprises about 5% to about 70% of a stearin shea fraction and about 30% to about 95% of an olein shea fraction.

9. The food product of claim 4-8, wherein the plant- or microbial-based oil is present in an amount between about 10% to about 30 % w / w of the food product.

10. The food product of any one of claims 1-9, wherein the food product is a cheese.

11. The food product of any one of claims 1-10, wherein the fermented non-dairy nutrient base is a fermented plant milk or a fermented microbial-based milk.

12. The food product of claim 11, wherein the fermented plant milk comprises (i) water, and (ii) milled seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof.

13. The food product of claim 11 or 12, wherein the fermented plant milk is selected from soybean milk, lupini bean milk, almond milk, hemp seed milk, melon seed milk, pumpkin seed milk, oat milk, pea milk, fava bean milk, chickpea milk, sunflower seed milk, edamame milk, lentil milk, pistachio milk, peanut milk, walnut milk, cashew milk, coconut milk, watermelon seed milk, and macadamia milk.

14. The food product of any one of claims 1-13, wherein the hydrocolloid fiber is selected from a fiber extract from seaweed, a fiber extract from an algae, acacia gum, locust bean gum, guar gum, pectin, cellulose or a cellulose derivative, konjac, alginate, carrageenan, gellan gum, agar, pullulan, dextran, curdlan, levan, and xanthan.

15. The food product of claim 14, wherein the algae is selected from Rhodophyceae algae, Phaeophyceade algae, or Chlorophyceae algae.

16. The food product of claim 14, wherein the hydrocolloid fiber is a fiber extract from a microorganism of a genus selected from Xanthomonas, Sphingomonas, Pseudomonas, Aureobasidium, Streptococcus, Leuconostoc, Acetobacter, Azotobacter, Pseudomonas, Alcaligenes, Bacillus, Zymomonas, Aerobacter, Acetobacter, Actinomyces, Erwinia, Rhanella, Lactobacillus, Microbacterium, and Serratia.

17. The food product of any one of claims 1-16, wherein the hydrocolloid fiber forms a synergetic gel.

18. The food product of any one of claims 1-17, wherein the hydrocolloid fiber is 1.0% w / w or less of the coagulated food product.

19. The food product of any one of claims 1-18, wherein the non-dairy protein is present in an amount greater than or equal to 20% w / w.

20. The food product of any one of claims 1-19, wherein a protein isolate or a protein concentrate was not added to the non-dairy nutrient base or the food product.

21. The food product of any one of claims 1-20, wherein the coagulated food comprises only whole plant foods as ingredients.

22. The food product of any one of claims 1-21, wherein the coagulated food product consists of 8 or fewer ingredients.

23. The food product of any one of claims 1-20, wherein the food product has a complex modulus (G*) between about 30000 PA and about 50000 PA at a frequency of 2-20 HZ.

24. The food product of any one of claims 1-22, wherein the food product has a phase angle (A) between about 8 and about 16 at a frequency of 2-20 HZ.

25. The food product of any one of claims 1-23, wherein the food product has a cut force slope of less than 1.5 N / s.

26. The food product of any one of claims 1-24, wherein the food product is fully fermented.

27. The food product of any one of claims 1-25, wherein the fermented non-dairy nutrient base is fermented with mesophilic or thermophilic bacteria.

28. The food product of any one of claims 1-25, wherein the fermented non-dairy nutrient base is fermented with at least one bacteria selected from Lactococcus lactis biovar diacelylaclis. Leuconostoc mesenleroides. and Propionibacteria or a combination thereof.

29. A method of producing a food product, comprising:(a) inoculating a non-dairy nutrient base with at least one microbial culture;(b) adding at least one hydrocolloid fiber to the non-dairy nutrient base inoculated with the at least on microbial culture, thereby inducing coagulation and formation of a curd;(c) removing water from the curd such that protein content of the curd is at least two times higher than protein content of the non-dairy nutrient base of a w / w basis; and(d) aging the curd to allow fermentation of the non-dairy nutrient base by the with at least one microbial culture comprising at least one gas-forming bacteria; wherein fermentation with the at least one gas-forming bacteria results in eye formation in the food product.

30. The method of claims 29, wherein the at least one gas-forming bacteria is selected from Lactococcus lactis biovar diacelylaclis. Leuconostoc me senter aides, and Propionibacteria or a combination thereof.

31. A method of producing a food product, comprising:(a) homogenizing a non-dairy nutrient base;(b) inoculating the non-dairy nutrient base with at least one microbial culture;(c) adding at least one hydrocolloid fiber to the non-dairy nutrient base inoculatedwith the at least on microbial culture, thereby inducing coagulation and formation of a curd;(d) removing water from the curd such that protein content of the curd is at least two times higher than protein content of the non-dairy nutrient base of a w / w basis; and(e) aging the curd to allow fermentation of the non-dairy nutrient base by the with at least one microbial culture.

32. The method of claim 31, wherein homogenizing the non-dairy nutrient base comprises high sheer homogenization, high pressure homogenization, or a combination thereof.

33. The method of any one of claims 29-32 further comprising shaping the curd prior to aging the curd, wherein the shape is optionally a wheel, loaf, or a block.

34. The method of any one of claims 29-33 further comprising acidification or brining of the curd after removing water and prior to aging the curd.

35. The method of any one of claims 29-34 further comprising coating the curd with a water-permeable material immediately prior to aging the curd.

36. The method of any one of claims 29-35, wherein the curd shaped in a microperforated cheese mold.

37. The method of any one of claims 29-36, wherein aging the curd is performed at a relative humidity of between about 20% to about 85% and, optionally, a temperature between about 10°C to about 22°C.

38. The method of any one of claims 29-37 further comprising adding fermentation promotors or natural flavoring agents to the non-dairy nutrient base.

39. The method of claim 38, wherein the fermentation promotors are selected from sauerkraut, miso, and nutritional yeast.

40. The method of any one of claims 29-39 further comprising filtering the non-dairy nutrient base to remove solid particles prior to adding the at least one hydrocolloid fiber.

41. The method of any one of claims 29-40 further comprising contacting the non-dairy nutrient base with a protease, an amino peptidase, an amylase, a cellulase, a hemicellulose, or a combination thereof prior to adding the at least one hydrocolloid fiber.

42. The method of any one of claims 29-41, wherein the non-dairy nutrient base further comprises at least one plant- or microbial-based oil to the non-dairy nutrient base.

43. The method of claim 42, wherein the at least one plant- or microbial -based oil comprises a solid fat content (SFC) curve that is + / - 60% of a SFC curve of anhydrous milk fat (AMF) or lard at 10°C, + / - 60% of a SFC curve of AMF or lard at 15°C, + / - 60% of a SFC curve of AMF or lard at 20°C, + / - 50% of a SFC curve of AMF or lard at 25°C, + / - 50% of a SFC curve of AMF or lard at 30°C, + / - 20% of a SFC curve of AMF or lard at 35°C, + / - 10% of a SFC curve of AMF or lard at 40°C, or any combination thereof.

44. The method of claim 40 or 43, wherein the at least one plant- or microbial-based oil is selected from illipe fat, coco butter, sal butter, shea butter, shea olein, shea stearin, coconut oil, palm oil, palm kernel, pongamia oil, and any combination thereof.

45. The method of any one of claims 29-44, wherein the at least one microbial culture comprises a thermophilic bacterial starter culture or a mesophilic bacterial starter culture.

46. The method of anyone of claims 29-45, wherein the at least one microbial culture comprises a lactic acid bacteria, a propionic acid bacteria, an acetic acid bacteria, or a yeast.

47. The method of any one of claims 29-46, wherein the curd after removing water has a protein content of greater than or equal to 12% w / w, greater than or equal to 15% w / w, or greater than or equal to 20% w / w.

48. The method of any one of claims 29-47, wherein the non-dairy nutrient base has a pH of 5.2 or less prior to adding the at least one hydrocolloid fiber.

49. The method of any one of claims 29-48, wherein the non-dairy nutrient base is a plant-based milk.

50. The method of claim 49, wherein the plant milk comprises (i) water, and (ii) milled seeds, milled nuts, milled beans, milled grains, milled vegetables, or a combination thereof.

51. The method of any one of claims 29-50, wherein the non-dairy nutrient base comprises at least 60%, at least 65%, at least 70% at least 75%, or at least 80% w / w of water when the non-dairy nutrient base is inoculated with the at least one microbial culture.

52. The method of any one of claims 29-51, wherein the non-dairy nutrient base is selected from soybean milk, lupini bean milk, almond milk, hemp seed milk, melon seed milk, pumpkin seed milk, oat milk, pea milk, fava bean milk, chickpea milk, sunflower seed milk, edamame milk, lentil milk, pistachio milk, peanut milk, walnut milk, cashew milk, coconut milk, watermelon seed milk, and macadamia milk.

53. The method of any one of claims 29-52, wherein the hydrocolloid fiber forms a synergetic gel.

54. The method of any one of claims 29-53, wherein the hydrocolloid fiber is selected from a fiber extract from seaweed, an algae, acacia gum, locust bean gum, guar gum, pectin, cellulose or a cellulose derivative, konjac, alginate, carrageenan, gellan gum, agar, pullulan, dextran, curdlan, levan, and xanthan.

55. The method of claim 54, wherein the algae is selected from Rhodophyceae algae, Phaeophyceade algae, or Chlorophyceae algae.

56. The method of claim 54, wherein the hydrocolloid fiber is a fiber extract from a microorganism of a genus selected from Xanthomonas, Sphingomonas, Pseudomonas, Aureobasidium, Streptococcus, Leuconostoc, Acetobacter, Azotobacter, Pseudomonas, Alcaligenes, Bacillus, Zymomonas, Aerobacter, Acetobacter, Actinomyces, Erwinia, Rhanella, Lactobacillus, Microbacterium, and Serratia.

57. The method of any one of claims 29-56, wherein the method does not comprise adding a protein isolate or protein concentrate to the non-dairy nutrient base, the curd, or both.

58. A method of solid-state fermentation, comprising: coagulating a non-dairy nutrient base by contacting the non-dairy nutrient base with at least one hydrocolloid fiber to form a solid or gel and; inoculating the non-dairy nutrient base with at least one microbial culture comprising at least one gas-forming bacteria; and incubating the solid or gel to allow fermentation of the non-dairy nutrient base by the at least one microbial culture in a solid-state, thereby producing a non-dairy cheese comprising at least one eye.

59. The method of claim 58, wherein the at least one gas-forming bacteria is selected from Lactococcus lactis biovar diacelylaclis. Leuconostoc me senter aides, and Propionibacteria or a combination thereof.

60. A method of solid-state fermentation, comprising: homogenizing a non-dairy nutrient base; coagulating the non-dairy nutrient base by contacting the non-dairy nutrient base with at least one hydrocolloid fiber to form a solid or gel and; inoculating the non-dairy nutrient base with at least one microbial culture; and incubating the solid or gel to allow fermentation of the non-dairy nutrient base by the at least one microbial culture in a solid-state, thereby producing a non-dairy cheese that is capable of melting.

61. The method of claim 60, wherein homogenizing the non-dairy nutrient base comprises high sheer homogenization, high pressure homogenization, or a combination thereof.

62. The method of any one of claims 58-61, wherein during incubation, the non-dairy nutrient base is dehydrated at a dehydration rate consistent with a dehydration rate of dairy milk when incubating dairy cheese.

63. The method of any one of claims 58-62, wherein during incubation, the non-dairy nutrient base is acidified at an acidification rate consistent with an acidification rate of dairy milk when incubating dairy cheese.

64. The method of any one of claims 58-63, wherein the at least one microbial culture comprises a mesophilic bacteria or a thermophilic bacteria.

65. A food product obtained by the methods of any one of claims 29-64.

66. The food product of claim 65, wherein the food product is a cheese.

67. The food product of claim 65 or 66, wherein the food product comprises only whole plant foods and the hydrocolloid fiber as ingredients.

68. The food product of any one of claims 65-67, wherein the food product consists of 8 or fewer ingredients.

69. The food product of any one of claims 65-68, wherein the food product has a protein content of greater than or equal to 15% w / w, or optionally greater than or equal to 20% w / w.

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