Plant-based cheese compositions and methods of making

Abstract

Plant prolamin-based cheese compositions are described, as are methods of making them.

Description

PLANT-BASED CHEESE COMPOSITIONS AND METHODS OF MAKINGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This is a PCT International Application that claims benefit to U.S. Provisional Patent Application No. 63 / 584,723 filed on September 22, 2023, which is herein incorporated by reference in its entirety.BACKGROUND

[0002] The present invention relates to plant-based dairy analog products, including plant-based cheese analog products and methods of making them.SUMMARY OF THE INVENTION

[0003] The present disclosure is based, at least in part, on a plant-based cheese product made from a plant prolamin isolate. In one aspect, plant-based cheese product comprising less than 10% by weight of a plant prolamin isolate, combined with a fat and a structural component in water, to yield a cheese product comprising a prolamin non- covalent network. In one aspect, the plant-based cheese product comprises no more than 4%, nor more than 5%, no more than 6%, no more than 7%, no more than 8%, or no more than 9% by weight of the plant prolamin isolate. In another aspect, the plant prolamin isolate is obtained from corn. In yet another aspect, the plant prolamin isolate comprises a zein or a functional analog thereof. In yet another aspect, the plant prolamin isolate comprises a corn zein. In one aspect, the plant prolamin isolate is substantially free of carotenoids. In one aspect, the present disclosure provide a plant-based cheese product made of plant prolamin isolate, and such plant prolamin comprises about 75%, about 80%, about 81 %, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90% or about 91 % by weight prolamin protein, about 2% to about 20% by weight fat, and the balance by weight moisture. In one aspect, the prolamin protein is a zein. The cheese product has a pH in the ranges of 2.9 - 3.1 , 2.95 - 3.05, or 2.96 - 3.04. In another aspect, the plant-based cheese product also comprises about 0.1 - 25% by weight of the fat, and 1 -30% by weight the structural component. In another aspect, the fat is selected from the group consisting of sunflower oil, canola oil, safflower oil, soybean oil, avocado oil, olive oil, corn oil, flaxseed oil, almond oil, coconutoil, peanut oil, pecan oil, cottonseed oil, palm oil, palm stearin, palm olein, palm kernel oil, rice bran oil, sesame oil, cocoa butter, shea butter, shea stearin, shea olein, palm kernel stearin, palm kernel olein, grape seed oil, hazelnut oil, Brazil nut oil, linseed oil, acai palm oil, passion fruit oil and any combination thereof. In another aspect, the fat comprises an oleic acid content of greater than 20% by weight. In another aspect, the structural component comprises a thickening agent and / or a gelling agent. In another aspect, the structural component is selected from the group consisting of a starch, a microbial or vegetable gum, a protein, a sugar polymer, and any combination thereof. In yet another aspect, the structural component is selected from the group consisting of arrowroot, cornstarch, katakuri starch, potato starch, sago, wheat flour, almond flour, tapioca, tapioca flour, starch derivatives, modified or pre-gelatinized starches, alginin, guar gum, locust bean gum, gellan gum, tara gum, gum Arabic, Konjac gum, xanthan gum, collagen, agar, carboxymethyl cellulose, pectin, carrageenan, and any combination thereof.

[0004] In one aspect, the present disclosure provides a plant-based cheese product further comprises a plasticizer. In one aspect, the plasticizer is selected from water, a food grade acid, glycerol, a sugar, and any combination thereof. In another aspect, the food grade acid is selected from the group consisting of carboxylic acid, gluconic acid, citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid and any combination thereof. In yet another aspect, the plasticizer comprises or consists of a combination of a food grade acid and water. In yet another aspect, the food grade acid is lactic acid or gluconic acid. In one aspect, the plasticizer in an amount of up to 5% by weight.

[0005] Another aspect of the present disclosure provides a method of making a plantbased cheese product. The steps comprise (i). combining the structural component, such as starch, with fat, such as oil, and water; heating at a temperature of at least about 85°C, thereby forming a mixture having a viscosity that permits formation of a non-covalent prolamin network at a temperature greater than the glass transition temperature of the prolamin; (ii). combining the plant prolamin isolate with the plasticizer to produce a plasticized plant prolamin composition; (iii).at the temperature of (i), combining the mixture of (i) with the plasticized plant prolamin composition of (ii), to form a cheese mixture; and (iv). cooling the cheese mixture to a temperature from about 65°C to about75°C for at least 3 minutes, thereby providing a pasteurized plant-based cheese product. In one aspect, the heating in (i) is to a temperature from about 85°C to about 95°C. In another aspect, the plant prolamin has a glass transition temperature of greater than 40°C. In another aspect, the cooling in (iv) is to about 65 °C for at least 30 minutes. In one aspect, the plant prolamin is a zein. In another aspect, the zein is a corn zein. In yet another aspect, the plant prolamin isolate is substantially free of carotenoids. In yet another aspect, the plant prolamin isolate comprises about 75%, about 80% or about 90% by weight zein protein, about 2% to about 20% by weight fat, and the balance by weight moisture. In one aspect, the plant-based cheese product has a pH of 2.9 - 3.1 , 2.95 - 3.05, or 2.96 - 3.04. In another aspect, the plasticizer is selected from water, a food grade acid, glycerol, a sugar, and any combination thereof. In yet another aspect, the food grade acid is selected from carboxylic acid, gluconic acid, citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid, and any combination thereof. In yet another aspect, the plasticizer comprises or consists of a combination of a food grade acid and water. In one aspect, the food grade acid is lactic acid or gluconic acid.

[0006] Yet another aspect of the present disclosure provides a plant-based cheese product comprising water and a food grade acid in combination with less than 10% by weight corn zein, wherein the com zein is plasticized by the water and the food grade acid. In one aspect, the food grade acid is selected from carboxylic acid, gluconic acid, citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid, and any combination thereof. In another aspect, the food grade acid is lactic acid or gluconic acid. In yet another aspect, the plant-based cheese product exhibits 3.5 to 7.5 inches of stretch when subjected to an industry standard Fork Stretch test. In yet another aspect, the plantbased cheese a melt spread of at least 25% to about 50% when subjected to an industry standard oven melt spread test. In yet another aspect, the plant-based cheese exhibits a AE of less than 3.0 as compared to a positive control sample. In yet another aspect, the corn zein is substantially free of carotenoids.

[0007] The foregoing is intended to be illustrative and is not meant in a limiting sense. Many features and sub-combinations of the present disclosure may be made and will be readily evident upon a study of the following specification and accompanying drawings comprising a part thereof. These features and sub-combinations may be employedwithout reference to other features and sub-combinations.BRIEF DESCRIPTION OF THE DRAWINGS

[0008] In order to describe the manner in which the aspects and features of the present disclosure can be obtained, reference is made to the appended drawings. Understanding that these drawings depict only exemplary aspects of the disclosure and are not therefore to be considered to be limiting of its scope, the principles herein are described and explained with additional specificity and detail through the use of the accompanying drawings.

[0009] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0010] FIG. 1 is a series of color photographs of select samples of zein isolates as described herein.

[0011] FIG. 2 is a series of color photographs of different samples of melted plant-based cheese products prepared as described herein, each using a different zein isolate as described herein.

[0012] FIGs. 3A-3I is a series of bar graphs and tables summarizing properties of zein isolate samples tested. FIG. 3A is a bar graph of a color analysis of different zein isolate samples. FIG. 3B is a bar graph of bulk density of the different zein isolate samples. FIG. 3C is a bar graph of water activity of the different zein isolate samples. FIG. 3D is a bar graph of stretch and melting properties of different plant-based cheese products made using the different zein isolate samples. FIG. 3E is a table summarizing assessment of performance of the different plant-based cheese products according to which zein isolate was used. FIG. 3F is a table summarizing certain specifications of decolorized zein isolates used to prepare plant-based cheese product according to the present disclosure. FIG. 3G is a table summarizing results of water holding capacity testing of different zein isolate samples. FIG. 3H is a table summarizing results of oil holding capacity testing of of different zein isolate samples. FIG. 3I is a table summarizing results of surface hydrophobicity testing of different zein isolate samples.

[0013] FIGs. 4A-4C are a series of illustrations describing a process for making a plantbased cheese product using a corn zein isolate according to the present disclosure. FIG. 4A is a flow diagram of a process for making a plant-based cheese product using a corn zein isolate according to the present disclosure. FIG. 4B is a series of color photographs showing various aspects of a process as described herein and using a Thermomix. FIG. 4C is a series of color photographs showing various aspects of a process as described herein, performed in a Stephan Cooker.

[0014] FIGs. 5A-5B are two series of photographs of a plant-based cheese product made using a corn zein isolate as described herein. FIG. 5A is a series of color photographs showing melt performance of a plant-based cheese product made using a corn zein isolate according to the present disclosure. FIG. 5B is a series of color photographs of shreds prepared from different samples of plant-based cheese products made using different zein isolates as described herein.

[0015] FIGs. 6A-6B are bar graphs showing the impact of different zein incorporation methods on the performance of plant-based cheese products made according to the present disclosure. FIG. 6A is a bar graph of results of a Fork Stretch Test of different samples of plant-based cheese products made using different methods for incorporating a zein isolate, according to the present disclosure. FIG. 6B is a series of bar graphs showing results of Fork Stretch Testing of different samples of a plant-based cheese made using different zein isolates, after heating in an oven (left panel) or a microwave (right panel, performed on TOSHIBA Microwave Oven, MODEL EM925A5A-SS with Input of 120V~60Hz and1350 W 2450 MHz), compared to standard results for Fork Stretch of standard Commercial #1 brand cheese and a standard Commercial #2 cheese sample (middle panel).

[0016] FIG. 7 is a bar graph of the impact of zein content on the melt stretch of a plantbased cheese product made according to the present disclosure.

[0017] FIGs. 8A-8C are flow diagrams of processes tested according to the present disclosure. FIG. 8A is a flow diagram of a processing method for preparing a plant-based cheese product using a zein isolate, resulting in an unpasteurized product. FIG. 8B is a flow diagram of an improved process for preparing a plant-based cheese product using azein isolate as described herein. FIG. 8C is a flow diagram showing different pasteurization conditions tested by the present disclosure.

[0018] FIGs. 9A-9N is a series of photographs and drawings showing various test results from testing plant-based cheese products as described herein, using different pasteurization conditions. FIG. 9A shows the instruments used for cheese characterization. FIG. 9B is a series of photographs of shreds made of different samples of plant-based cheese made under different pasteurization processes as described herein. FIG. 9C is a bar graph of results of Fork Stretch testing of plant-based cheese product made under different pasteurization processes. FIG. 9D illustrates the melt features of the plant-based cheese. FIG. 9E illustrates the oven melt spread features of the plant-based cheese products. FIG. 9F illustrates the hardness of plant-based cheese products. FIGs. 9G-9I provides a color analysis of different samples of plant-based cheese products. FIG. 9J provides cohesiveness of the plant-based cheese made under different pasteurization conditions. FIG. 9K illustrates gumminess of samples of plantbased cheese products made under different pasteurization conditions. FIG. 9L provides sensory features of the plant-based cheese products made under different pasteurization conditions. FIGs. 9M-9N summarize the impact of different pasteurization conditions on the plant-based cheese products.

[0019] FIG. 10 is a flow chart of a process for making a plant-based cheese according to the present disclosure, including pasteurization.

[0020] FIGs. 11A-11 H is a series of photographs and graphs showing features and properties of plant-based cheese products made under different pasteurization conditions. FIGs. 11 A and 11 B are respectively a bar graph of a color analysis of different samples plant-based cheese product made by a process using different pasteurization as described herein, and a series of color photographs showing the color and melt performance of the different samples compared to a control sample. FIG. 11C is a bar graph of stretch and melt test results of different samples of plant-based cheese product made using different pasteurization conditions. FIG. 11 D is a series of photographs showing melting performance of different samples of plant-based cheese product made using different pasteurization conditions. FIG. 11E is a table summarizing results ofperformance testing of different samples of plant-based cheese product made using different pasteurization conditions. FIG. 11 F is a bar graph of results of Fork Stretch testing of different samples of plant-based cheese product made using different pasteurization conditions, following heating by microwave oven. FIG. 11G is a bar graph of results of Melt Spread testing of different samples of plant-based cheese product made using different pasteurization conditions, following heating in a regular oven. FIG. 11 H is a series of color photographs showing melt-spread performance of different samples of plant-based cheese product made using different pasteurization conditions

[0021] The drawings are non-limiting, and are not necessarily to scale, emphasis instead being placed on clearly illustrating principles of certain aspects of the present disclosure.DETAILED DESCRIPTION

[0022] The following detailed description references the accompanying drawings that illustrate various aspects and aspects of the present disclosure. The drawings and description are intended to provide exemplary aspects and aspects of the present disclosure rather than limiting the scope. Other equivalents, variants or changes can be made without departing from the essence or spirit of the present disclosure. The following description is, therefore, not to be taken in a limiting sense.

[0023] The present disclosure provides plant-based cheese products made from plant- derived ingredients, substantially free of real dairy ingredients such as any animal milks and / or animal proteins. The plant-based cheese products disclosed herein include for example less than 10% by weight of a plant prolamin isolate, a fat, a structural component and optionally water. Without wishing to be bound by theory, a plant prolamin isolate combined with the fat and the structural component according to the amounts and process as described herein, yields a non-dairy cheese product comprising a prolamin non-covalent network. The plant-based cheese products described herein exhibit at least one or more performance features or properties comparable to that of a real dairy cheese, for example, stretch, rheological melting profile, loss of shape / melting characteristics, and / or hardness.I. Terminology

[0024] The phraseology and terminology employed herein are for the purpose of description and should not be regarded as limiting. For example, the use of a singular term, such as, “a” is not intended as limiting of the number of items. Also, the use of relational terms such as, but not limited to, “top,” “bottom,” “left,” “right,” “upper,” “lower,” “down,” “up,” and “side,” are used in the description for clarity in specific reference to the figures and are not intended to limit the scope of the present inventive concept or the appended claims.

[0025] Further, any one of the features of the present inventive concept may be used separately or in combination with any other feature. References to the terms “aspect,” “aspects,” and / or the like in the description mean that the feature and / or features being referred to are included in, at least, one aspect of the description. Separate references to the terms “aspect,” “aspects,” and / or the like in the description do not necessarily refer to the same aspect and are also not mutually exclusive unless so stated and / or except as will be readily apparent to those skilled in the art from the description. For example, a feature, structure, process, step, action, or the like described in one aspect may also be included in other aspects but is not necessarily included. Thus, the present inventive concept may include a variety of combinations and / or integrations of the aspects described herein. Additionally, all aspects of the present disclosure, as described herein, are not essential for its practice. Likewise, other systems, methods, features, and advantages of the present inventive concept will be, or become, apparent to one with skill in the art upon examination of the figures and the description. It is intended that all such additional systems, methods, features, and advantages be included within this description, be within the scope of the present inventive concept, and be encompassed by the claims.

[0026] Any term of degree such as, but not limited to, “substantially” as used in the description and the appended claims, should be understood to include an exact, and / or a similar, but not exact configuration or amount. For example, “substantially planar surface” means having an exact planar surface and / or a similar, but not exact planar surface. Such similar planar surface, though not strictly planar, is not departure from aplane in a significant way, such that it still bears the basic feature of a planar surface. Similarly, “substantially free of A” or “substantially absence of A” includes 0% of A, and / or any reasonable insignificant amount of A. For example, such insignificant amount may be less than 10%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, less than 1.5%, less than 1.0%, less than 0.8%, less than 0.6%, less than 0.5%, less than 0.4%, less than 0.3%, less than 0.2%, less than 0.1 %, less than 0.05%, less than 0.02%, or about 0%, and such percentage can be any reasonable percentage measure, such as weight percentage, molar percentage, or weight to volume percentage.

[0027] The terms “about” or “approximately,” as used in the description and the appended claims, should be understood to include the recited values or a value that is three times greater or one third of the recited values. For example, about 3 mm includes all values from 1 mm to 9 mm, and approximately 50 degrees includes all values from 16.6 degrees to 150 degrees. For example, they can refer to less than or equal to ± 5%, such as less than or equal to ± 2%, such as less than or equal to ± 1 %, such as less than or equal to ± 0.5%, such as less than or equal to ± 0.2%, such as less than or equal to ± 0.1 %, such as less than or equal to ± 0.05%.

[0028] The terms "comprising," "including" and "having" are used interchangeably in this disclosure. The terms "comprising," "including" and "having" mean to include, but not necessarily be limited to the things so described.

[0029] Lastly, the terms “or” and “and / or,” as used herein, are to be interpreted as inclusive or meaning any one or any combination. Therefore, “A, B or C” or “A, B and / or C” mean any of the following: “A,” “B” or “C”; “A and B”; “A and C”; “B and C”; “A or B”; “A or C”; “B or C”, “A, B and C.” An exception to this definition will occur only when a combination of elements, functions, steps, or acts are in some way inherently mutually exclusive.II. Ingredients

[0030] Unless otherwise indicated, all ingredients used in the present disclosure are food grade ingredients. In some aspects, the ingredients are used in the amount that areclassified as GRAS by FDA. "GRAS" stands for the phrase Generally Recognized as Safe. Under sections 201 (s) and 409 of the Federal Food, Drug, and Cosmetic Act (the Act), any substance that is intentionally added to food is a food additive, that is subject to premarket review and approval by FDA, unless the substance is generally recognized, among qualified experts, as having been adequately shown to be safe under the conditions of its intended use, or unless the use of the substance is otherwise excepted from the definition of a food additive.(i) Plant prolamin isolates

[0031] Prolamins are a group of plant storage proteins characterized by a high glutamine and proline content. They are usually found in plants, mainly in the seeds of cereal grains, such as wheat, barley, rye, corn, sorghum, and oats. They have poor solubility in water. The plant-based prolamins for use in the present disclosure include, but are not limited to, prolamin proteins from a cereal grain plant such as wheat, barley, rye, corn, sorghum, oats, and their genetically modified species.

[0032] A protein isolate is a concentrated form of protein isolated from a food source, including but not limited to a plant, and which may include other components such as fat and / or water, but consists primarily of proteins. A prolamin isolate is a prolamin extracted and isolated from one or more plant sources and may be purified and / or modified for use as one of the ingredients in making the plant-based cheese products of the present disclosure. Plant sources for prolamin include but are not limited to wheat, barley, rye, corn, sorghum, oats, and their genetically modified species. Prolamins include, but are not limited to, gliadin, hordein, secalin, zein, kafirin, avenin, gluten, or any combination thereof. In certain aspects, a plant prolamin isolate as described herein comprises about 75%, about 80%, about 81 %, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90% or about 91 % by weight of prolamin protein, about 2% to about 20% by weight of fat, and the balance by weight moisture. In other aspects, a plant prolamin isolate described herein comprises 75%, 80%, 81 %, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90% or 91 % by weight of a zein protein. In various aspects, a plant prolamin isolate is a corn zein isolate.

[0033] A plant prolamin isolate also encompasses functional analogs or equivalentvariants that result naturally from alternative splicing during transcription or from genetic coding differences and may retain significant sequence homology with a wild type prolamin, e.g., at least about 70% sequence homology, preferably at least about 80% sequence homology, and more preferably at least about 90% or greater sequence homology. Alternatively, or additionally, such functional analogs or equivalent variants may result from non-naturally occurring synthetic alterations made to a natural prolamin to bestow on the variant more desirable characteristics for use in the present cheese product, for example, enhanced ability to form a non-covalent network. In one aspect, the functional analog may be a prolamin analogue that incorporates one or more amino acid substitutions, additions, or deletions. Amino acid additions or deletions include both terminal and internal additions or deletions. Examples of suitable amino acid additions or deletions may include those that occur at positions within the protein that are not closely linked to activity, or additions or deletions at the N- or C- terminus of the protein. Amino acid substitutions may include conservative amino acid substitutions such as the substitution of a non-polar (hydrophobic) residue such as alanine, isoleucine, valine, leucine or methionine with another non-polar (hydrophobic) residue; the substitution of a polar (hydrophilic) residue with another such as between arginine and lysine, between glutamine and asparagine, between glutamine and glutamic acid, between asparagine and aspartic acid, and between glycine and serine; the substitution of a basic residue such as lysine, arginine or histidine with another basic residue; or the substitution of an acidic residue, such as aspartic acid or glutamic acid with another acidic residue.

[0034] In one aspect, a plant prolamin isolate is a zein isolated from maize, i.e., corn plants. As used herein the term “zein” encompasses a prolamin isolated from corn having a molecular weight in the range of from 20 to 30 kDa, preferably, 22 to 25 kDa, composed of a high amount of hydrophobic amino acids, such as glutamine, leucine, proline and alanine, e.g., greater than 50% hydrophobic amino acid content, and functionally equivalent variants thereof as described above. Zein as used herein may comprise a mixture of zein proteins, e.g., a mixture of proteins with various molecular size, solubility, and charge such as a mixture of two or more of a-zein, p-zein, y-zein and 5-zein, or may comprise a single zein protein, provided that the mixture comprises minimal cysteine residues to minimize or prevent the formation of a covalent linked network, e.g. 1 or 2cysteine residues per zein subunit. In one aspect, the zein protein comprises primarily a- zein, e.g., greater than 85% a-zein. In another aspect, zein may comprise a functional analog of a-zein, p-zein, y-zein or 5-zein, or any mixture thereof. Such functional analog may incorporate conservative amino acid substitutions, substitution with derivatized amino acids, or amino acid additions or deletions, for example at the termini thereof. In another aspect, the zein protein comprises primarily a-zein and (3-zein, e.g., greater than 85% of the two, or a functionally equivalent variant thereof. In yet another aspect, the zein protein comprises less than 10% of y-zein, preferably less than 8%, 5% or 2%. In yet another aspect, the zein protein comprises less than 10% of 5-zein, preferably less than 8%, 5% or 2%.

[0035] In certain aspects, a prolamin isolate, such as a zein isolate used in the present disclosure is substantially free of carotenoids. Carotenoids are a class of more than 750 naturally occurring pigments synthesized by plants, algae, and photosynthetic bacteria. These richly colored molecules account for the yellow, orange, and red colors of many plants. Maize contains considerable amounts of carotenoid pigments, such as zeaxanthin, lutein and provitamin A carotenoids, [3-carotene, a-carotene and [3- cryptoxanthin. In the present disclosure, it is desirable to remove carotenoids, such as removal of at least 80% of carotenoids. Preferably at least 85%, 88%, 85%, 90%, or 95% of carotenoids are removed. In one aspect, the zein isolates have significantly reduced amount of lutein or zeaxanthin, such as reduced to about 20%, about 15%, about 10%, about 8%, about 5% or about 4% of lutein or zeaxanthin found in the natural zein. In another aspect, the zein isolates are substantially free of carotenoids. The term “substantially free of carotenoids” encompasses a composition containing no carotenoids, or containing total carotenoids in an amount less than 30 pg / g, preferably less than about 28 pg / g, less than about 27 pg / g, less than about 25 pg / g, less than about 24 pg / g, less than about 23 pg / g, less than about 22 pg / g, about 21 pg / g, less than about 20 pg / g, less than about 18 pg / g, about 15 pg / g, less than about 12 pg / g, less than about 10 pg / g, about 9 pg / g, less than about 8 pg / g, less than about 7 pg / g.

[0036] A plant prolamin isolate, such as a zein isolate obtained from corn may be obtained through extraction. For example, a prolamin isolate may be obtained by ethanol extraction of gluten corn meal, optionally followed by additional carotenoid removalprocesses. Zein isolates are also commercially available for example from Flo Chemical Corp, which supplies “FloZein Products” made by ethanol extraction of corn gluten meal followed by carbon filtration. Such zein isolate may contain total carotenoids up to about 130 pg / g, including over 50 pg / g of lutein and over 50 pg / g of zeaxanthin. These commercial zein products may be subjected to further carotenoid removal treatments, such as ethanol wash, to significantly reduce their carotenoid contents. Such carotenoid removed zein isolates are termed as “decolorized zein” or “decarotenoided zein”. A desired zein isolate may comprise at least 75% by weight of protein, about 2 to about 20% by weight of fat, and at least 2% moisture. Preferably, the zein isolate comprises about 80% to about 92% by weight of protein, 2% to about 14% by weight of fat, and up to about 8% moisture. Specifically, a zein isolate as used herein may possess about 90% of protein, about 2% of fat, and about 7% of moisture, or 90% of protein, 2% of fat, and the balance moisture. Additionally, or alternatively, when such zein isolate is used in the plant-based cheese product, such obtained cheese product may be acidic, such as with a pH less than 4, such as about 3.00, about 2.95, about 2.96, about 2.98, or about 2.99. Additionally, or alternatively, the zein isolate may have a bulk density (4.5 g in cm3) above 30, preferably above 40. Specifically, such bulk density is around 33, around 39, around 40, or around 47, or when measured by g / ml, the bulk density is in the range of about 0.09 g / ml to about 0.15 g / ml. Additionally, or alternatively, such zein isolate may be in particle or powder form. A preferred particle size may be around 1 mm. On the other hand, particles of zein isolate may have a size distribution. For example, the zein isolate particles may have 22% passing through Mesh #8, about 66% passing through Mesh #16 (1 mm sieve), and about 14% passing through Mesh #18. Additionally, or alternatively, the zein isolate possess preferred color, such as measured by Hunter Color L, a and b The zein isolate may have a Hunter Color L more than 69, such as in the range of about 70 to about 85, such as about 70, about 71 , about 72, about 73, about 74, about 75, about 76, about 77, about 78, about 79, about 80, about 81 , about 82, about 83, about 84, or about 85. In one aspect, the Hunter Color L is 70, 71 , 72, 73, 74, 75, 76, 77, 77.8, 78, 79, 80, 81 , 82, 83, 84, or 85. The zein isolate may have a Hunter Color a being less than 5.6, or in the range of about 4 to about 12, such as about 4.5, about 5.4, about 7.2, about 10.2, about 11 .4, or about 12.8. In one aspect, the zein isolate may have Hunter Color avalue as 4.5, 5.4, 7.2, 10.2, 11.4, or 12.8. The zein isolate may have a Hunter Color b being less than 40.5, or in the range of about 33.0 to about 62.0, such as about 33.0, about 34.0, about 42.0, about 45.0, about 46.0, about 57.0 or about 62.0. In one aspect, the zein isolate may have Hunter Color b value as 33.0, 33.9, 42, 45, 46, 57 or 62. Additionally, or alternatively, the zein isolate may have a water holding capacity close to about 100%. Additionally, or alternatively, the zein isolate may have an oil holding capacity over about 700%. Additionally, or alternatively, the zein isolate may have a surface hydrophobicity. Such surface hydrophobicity when measured by Index of Surface Hydrophobicity (So) may be over 4,000 So, such as from 4,000 So to 5,000 So. Specifically, the zein isolate may contain about 90% by weight of protein, about 2% by weight of fat, about 7% of moisture, and with a pH of about 2.96, a bulk density about 47, a Hunter Color L at about 77.8, Hunter Color a at 5.4, Hunter Color b at 33.9, oven melt / stretch at 131.5 mm, and microwave melt / spread at 125.5.

[0037] The prolamin isolate used in the plant-based cheese products is present in an amount that provides stretchability comparable to that of dairy cheese, when measured by industry standard stretch tests as the Fork Stretch test as described in the examples herein. Stretchability is based on the extensional rheology of the product, and the extensional flow of the product in response to the application of a linear pulling force (i.e., stretching) in a particular direction. In one aspect, a prolamin cheese product as described herein is stretchable in an amount of about 100% in a linear direction from a resting or baseline position, i.e., a position in which the product is not stretched or under any force or tension, without breaking at a temperature within the range of about 40°C to about 80°C. Preferably, the plant-based cheese products of the present disclosure are stretchable in a linear direction in an amount of greater than 100% from a resting position, such as an amount of at least 200%, 300%, 400%, 500%, or more at a temperature of about 50°C to about 55°C. Stretchability may also be expressed as a length. In another aspect, the plant-based cheese product is stretchable to at least double its length in a resting position, and preferably to at least about 2 or 3 times its length at rest, without breaking, when tested at a temperature of about 40°C to about 80 °C, e.g., at about 50°C. In this regard, the amount of plant prolamin isolate in the product will vary with the amounts of the other ingredients in the product. As disclosed herein, the amount of theprolamin isolate is no more than 10% by weight, (i.e., wt%) of the cheese product, and in various aspects is no more than 4%, no more than 5%, no more than 6%, no more than 7%, no more than 8%, or no more than 9% by weight of the cheese product.

[0038] Without being bound by theory, the amount of prolamin isolate used in the disclosed plant-based cheese products is believed to be sufficient to form a non-covalent linked protein network, similar to that formed by casein in natural cheese. The non- covalent linked network is formed by hydrophobic interactions and hydrogen bonding as opposed to covalent disulfide linkages. The formation of this non-covalent network is believed to provide the unexpected stretchability of the presently disclosed products, comparable to that of dairy cheese. The plant prolamin isolates used in the present cheese product are able to form networks that exhibit storage moduli (G1) greater than loss moduli (G"), i.e., G' > G", such that, for example, the tan 5 (G" / G') is greater than 0.5 but less than 1 .0.(ii) Plant-based Fats or Oils

[0039] The plant-based cheese products of the present disclosure also comprise a food grade fat or oil, preferably a plant-based edible fat or oil. Such fat or oil component may provide the plant-based cheese product with a desired melting behavior, in an amount that may be adjusted or modified based on the desired melting behavior of the cheese product. For example, to modify melting behavior of a product, a plant-based cheese product may include a fat or combination of fats which are solid at fridge temperatures (about 4°C-6°C), but which begin to melt when heated above normal ambient temperatures (e.g., about 20°C-24°C). Further, the fat content of the cheese may be altered in order to provide the plant-based cheese product mimicking a particular dairybased cheese type, such as imitating the texture, hardness and / or chewiness of the particular cheese. For example, a cheese product will comprise a greater amount of a solid fat to mimic a harder cheese such as Parmesan or Old Cheddar, while a cheese product that mimics a softer cheese, such as Brie, Camembert or Gorgonzola, will comprise a greater amount of a liquid (oil) fat. Altering the fat content may also be useful to produce a “low calorie” or “heart healthy” cheese product by providing a cheese productwith a lower fat content, or at least a lower saturated fat content. The plant-based cheese products made by the present disclosure may comprise about 0.1-25% by weight of fat and / or oil, preferably about 10% to about 25%, about 10% to about 20%, and more preferably from about 10% to about 18% by weight. In various aspects, plant-based cheese as described herein may comprise about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, or about 20% by weight of the fat or oil; or 10%, 11 %, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% by weight of the fat or oil.

[0040] The fat or oil may be selected from a saturated fat, an unsaturated fat, either monounsaturated or polyunsaturated, or any mixture thereof. The fat or oil may be a vegetable fat or oil. Such fat or oil may comprise but is not limited to sunflower oil, canola oil, safflower oil, soybean oil, avocado oil, olive oil, corn oil, flaxseed oil, almond oil, coconut oil, peanut oil, pecan oil, cottonseed oil, algal oil, palm oil, palm stearin, palm olein, palm kernel oil, rice bran oil, sesame oil, butteroil, cocoa butter, shea butter, shea stearin, shea olein, palm kernel stearin, palm kernel olein, grape seed oil, hazelnut oil, Brazil nut oil, linseed oil, acai palm oil, passion fruit oil, or any combination thereof. In some aspects, the fat comprises a combination of two or more fats, such as any combination of coconut oil, peanut oil, pecan oil, cottonseed oil, or palm oil. By way of non-limiting example, the fat may be provided by a commercially available product, such as Coconut oil Neutresca (Neutresca 51-25).(iii) Structural components

[0041] The plant-based cheese products of the present disclosure also comprise a structural component, in an amount of about 1 % to about 30% by weight. The structural component provides viscoelasticity while supporting the formation of the prolamin non- covalent networks within the matrix of the plant-based cheese product. Such matrix is preferably an aqueous matrix, and such structural component provides support, rather than inhibiting or preventing, the formation of non-covalent prolamin networks within such aqueous matrix. The structural component contributes to the structure of the product, and functions to retain moisture and emulsify the fat component(s) within the system. To effectuate the formation of non-covalent prolamin networks, the structural component willgenerally gel at a temperature at or below the glass transition temperature of the prolamin, such that the viscosity of the structural component on addition of prolamin at a temperature above its glass transition temperature allows the formation of the non- covalent prolamin network.

[0042] In one aspect, the structural component may comprise a thickening agent and / or a gelling agent. The thickening or gelling agent may comprise acacia (gum Arabic), agar, aluminum magnesium silicate, sodium alginate, sodium stearate, bladderwrack, bentonite, carbomer, carrageenan, Carbopol, xanthan gum, cellulose, microcrystalline cellulose (MCC), ceratonia, chitin, carboxym ethylated chitosan, chondrus, dextrose, furcellaran, gelatin, Ghatti gum, guar gum, hectorite, lactose, sucrose, maltodextrin, mannitol, sorbitol, honey, maize starch, wheat starch, rice starch, potato starch, gelatin, sterculia gum, xanthan gum, gum tragacanth, ethyl cellulose, ethyl hydroxyethyl cellulose, ethyl methyl cellulose, methyl cellulose, hydroxyethyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl cellulose, poly(hydroxyethyl methacrylate), oxypolygelatin, pectin, polygeline, povidone, propylene carbonate, methyl vinyl ether / maleic anhydride copolymer (PVM / MA), poly(methoxyethyl methacrylate), poly(methoxyethoxyethyl methacrylate), hydroxypropyl cellulose, hydroxypropyl methyl cellulose (HPMC), sodium carboxymethyl-cellulose (CMC), silicon dioxide, polyvinylpyrrolidone (PVP: povidone), or any combination thereof.

[0043] In another aspect, the structural component comprises a starch, a microbial or a vegetable gum, a protein, a sugar polymer, and any combination thereof. For example, the structural component may comprise arrowroot, cornstarch, katakuri starch, potato starch, sago, wheat flour, almond flour, tapioca, tapioca flour, starch derivatives, modified or pre-gelatinized starches, alginin, guar gum, locust bean gum, gellan gum, tara gum, gum Arabic, Konjac gum, xanthan gum, collagen, egg white, gelatin, agar, carboxymethyl cellulose, pectin, carrageenan, or any combination thereof.

[0044] In yet another aspect, the structural component comprises a combination of two or more structural components, such as a combination between or among tapioca, potato starch, corn starch, agar, pectin, Konjac gum, locust or xanthan gum. Such combination provides a desired viscoelasticity and permits the prolamin isolates, such as zein, to formnetworks via non-covalent linkages. The amount of each component may be varied depending on the properties of each component. For example, a combination of tapioca and corn starch at a weight ratio of about 2:1 tapioca to com starch may be used. This ratio may be altered in order to arrive at the desired properties of the final plant-based cheese product. Other combinations of thickening and / or gelling agents may also be utilized, for example, combinations of starch, such as tapioca or com starch, with a gum such as locust bean gum, agar with a gum such as Konjac gum, a combination of starches, or a combination of gums. The structure component(s) may use commercially available products, such as Tapioca Flour Homecraft Create 390 food starch PRECISA® 604, modified food starch N-Creamer 2000, carrageenan Ticaloid® 710 H Powder (COLL 710H).

[0045] In one aspect, the amount of structural component(s) in the plant-based cheese product may generally be an amount that provides a ratio of structural component to fat in the range of about 1 :2 to about 2:1 structural component to fat, such as from about 1 :2 to about 1 :1 , from about 2:3 to about 1 :1 , from about 1 :1 to about 3:2, from about 3:2 to about 2:1. Preferably, the weight ratio of structural component to fat is about 2:1 , about 5:3, or about 4:3. The structural component(s) may account to about 1 % to about 30% by weight of the plant-based cheese product, preferably in the range of about 10% to 30% by weight, about 15% to 30% by weight, about 20% to 30% by weight, or about 24% to 28% by weight.(iv) Plasticizers

[0046] In certain aspects, the plant-based cheese products may optionally include a plasticizer to increase the melt and stretchability of the products. Suitable plasticizers comprise food grade plasticizers including, but not limited to, water, a food grade acid, a glycerol, a sugar or any combination thereof. The food grade acid may comprise levulinic acid, palmitic acid, stearic acid, and oleic acid, or food grade carboxylic acids, such as lactic acid, gluconic acid, citric acid, malic acid, tartaric acid, acetic acid, oxalic acid, or any combination thereof. Glycerol may comprise triethylene glycol, ethylene glycol, a polyethylene glycol (PEG), a PEG conjugate, or a PEG derivative. The sugar may comprise fructose, galactose, glucose, or any mixtures thereof. The plasticizer may be sorbitol. The plasticizer may comprise a mixture of a food grade acid or acids with water,such as solutions of lactic acid, gluconic acid, citric acid, malic acid, tartaric acid, acetic acid, oxalic acid, or any combination thereof in water. The concentration of acid in such a solution may be above 70% by weight, such as at 88%. In various aspects, the plasticizer is lactic acid in water, or gluconic acid in water. The plasticizer or mixture of plasticizers may be present in the final cheese product in an amount of about 0% to about 5% by weight, preferably in the range of 0.2% to about 4% by weight, or preferably about 0.2%, about 0.75%, about 1.5%, or about 4% by weight.

[0047] Alternatively, increased melt and stretch in the plant-based cheese product may be achieved by partial hydrolysis of the prolamin isolate using methods well-known in the art, such as partially hydrolyzing with an acid or a base, followed by precipitation using a base or acid, respectively.

[0048] When a prolamin isolate is treated with a plasticizer, a plasticized prolamin isolated is formed. Such plasticized isolate may exhibit a melting profile in which G' and G" are reduced at elevated temperatures, e.g., temperatures of between about 40°C to about 80°C. At room temperature, G' and G" values are generally greater than 104 Pa, in the range of 104 to 106 Pa, and are reduced to less than 104 Pa as temperature is increased. Properties of this plasticized isolate may be altered by inclusion of one or more additional ingredients, such as fats, structuring components, flavorants, nutrients, fillers and others, as described herein.

[0049] When the plant prolamin isolate is a corn zein, the use of plasticizers with zein may decrease the glass transition temperature (Tg) of zein. Zein non-covalent networks are formed at temperatures above zein’s glass transition temperature (~40°C) and revert to brittle behavior when temperatures drop below this point. This is undesired for a plantbased cheese product that will be stored at cooler temperatures such as are typical of refrigerators. The addition of a plasticizer may enhance the melt / stretch property of zein. The zein isolate can be plasticized by a food grade carboxylic acid, such as citric acid, malic acid, tartaric acid. The functionality achieved by adding a plasticizer to zein can also be achieved by other methods, including by partial hydrolysis of zein. In summary, a plasticizer may be optionally used to decrease the rigidity of the final plant-based cheese products.(v). Other Ingredients: Flavorants, Colorants, Fillers, and Others

[0050] In certain aspects, cheese product may include additional ingredients including, but not limited to, flavorants, colorants, nutrients, fillers, preservatives, or antioxidants.

[0051] The plant-based cheese product of the present disclosure may optionally contain a flavorant. A flavorant also known as flavor (or flavour) or flavorant, is a food ingredient used to improve the taste or smell of food. It changes the perceptual impression of food as determined primarily by the chemoreceptors of the gustatory and olfactory systems. Such flavorant may include salt, sugar, spices, herbs and the like isolated from natural sources. Such flavorant may also be synthetic and may mimic flavors or tastes of naturally occurring counterparts. The flavorant may be a peptide, an amino acid, a fatty acid that provide a salty, sour, bitter, or sweet taste. The fatty acid may comprise butyric acid, lactic acid, or capric acid which provides characteristic cheese flavor. The flavorant used may include dairy fat flavor, natural milk flavor, cheese taste, natural Monterey Jack Ku, natural Monterey Jack emulsion, natural Queso Crema in paste, natural masking flavor for corn protein, protein masking flavor YP, masking flavor TS-840-145-7, or any combination thereof.

[0052] The present plant-based cheese product may also comprise a food preservative. The food preservative is added to the cheese product to prevent spoiling, maintain the cheese product's nutritional quality, and / or improve appearance. The food preservative used may include, but is not limited to, sodium benzoate, sodium and calcium propionate, sorbic acid, ethyl formate, and sulfur dioxide.

[0053] The present plant-based cheese product may also comprise an antioxidant. Examples of the antioxidant include, but are not limited to, ascorbic acid, tocopherols, butylated hydroxy anisole and propyl gallate.

[0054] The present plant-based cheese product may also comprise added nutrients. Added nutrients suitable may include but not limited to, vitamins, minerals, and / or protein isolates. Vitamins include vitamin A, C, E, K, D, thiamin (vitamin Bi), riboflavin (vitamin B2), niacin (vitamin B3), vitamin Be, folic acid (vitamin B9), vitamin B12, and any mixture thereof. Minerals may include calcium, phosphorus, magnesium, sodium, potassium, chlorine, iron, zinc, iodine, selenium, copper, and any mixture thereof. The protein isolatenutrient is preferably a protein isolate from plant source, such as pea protein, soy protein, corn protein, wheat protein, rice protein, canola protein, peanut protein, bean protein, lentil protein, pumpkin seed, rice, brown rice, peanut, almond, chia seed, flax seed, or any combinations thereof. The protein isolate nutrient may be non-hydrolyzed, partially hydrolyzed or hydrolyzed and may be in the form of an intact protein, amino acid or peptide.

[0055] Such additional ingredients may be optionally included in the disclosed plantbased cheese products in an amount from about 0.01 % by weight to about 5% by weight, preferably in the range of about 0.1 %-2% by weight of the product, or any of 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1 , 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 18. 1.9 or 2% by weight of the product.

[0056] The plant-based cheese product of the present disclosure may also contain a filler to provide volume / bulk to the cheese product while not impacting desired properties, such as rheological melting properties, hardness, stretch, shape, shredability and slice ability. Examples of suitable fillers for this purpose include, but are not limited to, consumable inert components such as microcrystalline cellulose, maltodextrin, dextrin, pea protein, soy protein, inulin, sugars, and mixtures thereof. The filler may be included in the cheese product in an amount of about 0.01 %-15% by weight, preferably about 0.05% -10% by weight.(vi) Water

[0057] Plant-based cheese products as described herein also include water in an amount of at least about 30% by weight, such as at least about 35%, at least about 40%, at least about 50%, 60%, 70% or more. Preferably the amount of water in the cheese product is in the range of about 40-60% by weight. In various aspects, water content in the plant-based cheese product is about 40%, about 41 %, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, or about 50% by weight; or 40%, 41 %, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, or 50% by weight. As one of skill in the art will appreciate, water content may vary with the desired characteristics of the final plant-based cheese product. For example, a softer cheese product will generally include a relatively greater amount of water (e.g., 50% by weight or more), while a hardercheese product will generally include relatively less water (e g., less than 50% by weight, such as 40-45% by weight).III. Characterization of Plant-Based Cheese Products

[0058] The present plant-based cheese products may be characterized in terms of various properties such as stretch, melt, hardness, color, flavor and the like, using any of many known, conventional techniques, or other techniques applied to functional characterization of cheese. As described in the examples herein below, certain conventional, industry-standard tests were performed on plant-based cheese products disclosed herein to evaluate suitability as a dairy cheese replacement. The Cheese Fork Stretch test is conventionally used to measure the stretchability of cheese and cheese analog products. Cheese samples are cut into a 5 mm thick / 39.5 mm diameter (block cheese) or weighed into 15 g portions (shredded cheese) and placed into a PYREX® dish or PYREX® Petri dish. The samples are covered with either aluminum foil or a glass Petri dish cover. The samples are heated in the oven at 450°F for 8 minutes or in the microwave for 15-45 seconds. In one aspect, the microwave oven used is TOSHIBA Microwave Oven, MODEL EM925A5A-SS, with input 120V~60Hz and 1350 W 2450 MHz. Samples are then cooled for 2 minutes. Using a ring stand to hold a 2-foot ruler upright, a fork is slid horizontally under the cheese sample (0.5”) into the center of the cheese and slowly pulls upwards at an angle of 45 degrees. The stretch recording is made at the height of the cheese right before strand breakage.

[0059] The Cheese Schreiber Melt Test is used to measure the meltability of cheese and cheese analog products. Cheese samples are cut into 5 mm thick / 39.5 mm diameter circles (block), or 15 g of shredded cheese was placed in the center of a PYREX® Petri dish or PYREX® dish. Using a caliper, the cross sections of the cheese samples are measured. The samples are covered with tin foil or a glass Petri dish cover and placed in an oven at 450°F for 5 minutes or at 400°F for 10 minutes or in a microwave heating for 15-45 seconds. The samples are then removed from heating and allowed to cool for 15 minutes. The diameter of the melted samples is measured at three cross sections (the three measurements are then averaged to obtain a singular value). Meltability iscalculated using the following formula: melted cheese diameter — raw cheese diameter Meltability = - - - - - 100 raw cheese diameter

[0060] The Prolamin Microwave stretch is performed by shredding the plant-based cheese product (15g) using a cheese box grater (largest hole size). The cheese shreds are arranged with a ring mold (95 mm) in an even layer on a 6 in porcelain plate. The ring mold is then removed, and the plate is placed into a microwave for 20 seconds. In one aspect, the microwave oven used is TOSHIBA Microwave Oven, MODEL EM925A5A-SS, with input 120V~60Hz and 1350 W 2450 MHz. The samples are removed and allowed to cool to about 85°C, measured using an Infrared thermometer. Once cooled, the samples are placed at the base of a ring stand paired with a 2-foot ruler and the fork is slid into the cheese sample and pulled from the center. Measurements are made at the point right before the final cheese strand breaks. Measurements are repeated in five replicates. For example, such measured oven melt / spread may be in the range of about 110 mm to about 143 mm, such as about 110 mm, about 111 mm, about 113 cm, about 128 mm, about 131 mm, about 132 mm, or about 142 mm. The microwave melt / spread may be in the range of about 123 mm to about 139 mm, such as about 123 mm, about 124 mm, about 125 mm, about 126 mm, about 132 cm, about 133 mm, about 131 mm, about 132 mm, about 138 mm, or about 139 mm.

[0061] The Cheese Stretch Texture Analyzer is used to measure the force required to stretch the cheese or cheese analog. The plant-based cheese products (15-20 g) are loaded into the well of the texture analyzer and the metal ring is placed over the cheese or cheese analog sample (ensure the probe is aligned with the Cheese Texture Analysis too, “TPA”). The samples are placed into the oven (400°F for 10 minutes). The samples are removed from the oven and placed into the texture analyzer. Start the test when the cheese sample begins to stretch. Once the strands begin to break, the test is stopped, and data collected. The test is repeated in triplicate.

[0062] The Cheese TPA (Texture Profile Analysis) is used to measure the firmness of the cheese product in block form. Cheese samples are cut into 1” x 1” x 1” cubes and refrigerated for 3 hours prior to testing. The cheese samples are loaded into the TPA, andthe test program starts to run. Data is collected and the test run in triplicates.

[0063] The Pizza Oven Stretch Test is performed by preheating a commercial conveyer belt pizza oven to 500°F. An 8” round pre-made pizza crust is placed on a pizza peel, an amount (45.2 g) of tomato sauce added over top, and then 85 g of shredded samples of the present plant-based cheese products added on top, taking care to leave about %” around the edge of the crust and distribute the shreds relatively evenly across the surface of the pizza. The pizza is transferred to the conveyer belt and cooked in the oven for 4-5 minutes. The pizza is removed from the belt using the pizza peel and allowed to cool for 2 minutes. Using a ring stand to hold a 2-foot ruler upright, a fork is slid horizontally under the cheese sample (0.5”) into the center of the cheese and slowly pulls upwards at a 45° angle. The stretch recording is made at the height of the cheese right before strand breakage. The process is repeated in triplicate. The cheese samples should be stretchable in an amount of at least about 100% in a linear direction from a resting or baseline position without breaking at a temperature between about 40°C to about 80° C, preferably at a temperature between about 50°C to about 60°C.

[0064] Additional characterization measures may include gumminess, chewiness, and / or springiness, which are obtained through the compression tests as shown in the equations below.Area under 2nd compression peak1) Gumminess = Hardness XArea under 1st compression peak. Time to reach peak during 2nd compression2) Chewiness = Gumminess X -Time to reach peak during 1st compression> . . Time to reach peak during 2nd compression j ) Springiness = -Time to reach peak during 1st compression

[0065] Using the functional tests described herein, the plant-based cheese products are evaluated against a dairy-based cheese. For example, at room temperature, the tan 5 (G" / G') of a cheese product should be in the range of 0.2 to 0.5, and increases on heating such that below 70°C, G' > G" ( tan 5 is > 0.2 and < 1 ), at about 70° C, G' = G" (tan 5 is 1 ), and above 70°C, G' < G" (tan 5 ranges from greater than 1 to about 2 at temperatures of about 70°C to about 100°C). Thus, the melting profile of the cheese product may besuch that tan 5 increases from about 0.2-0.5 at room temperature to about 2.0 as temperature is increased up to about 100°C. Alternatively, the melting profile of the cheese product is such that storage moduli (G') is greater than loss moduli (G"), and tan 5 (G" / G') is greater than 0.5 but less than 1 .0. In another aspect, a cheese product exhibits a decrease in hardness and loss of shape at increased temperature, e.g., a decreased hardness at 50°C as compared to hardness at 5°C to mimic the softening of cheese at increased temperatures. The extent of decreased hardness will vary with the type of cheese product (i.e. , a harder or softer cheese). Hardness is measured as the maximum force recorded during the first compression of a double compression cycle, taken at different temperatures to quantify softening that occurs. Preferably, the cheese product exhibits a decrease in hardness at increased temperatures, from a hardness of 2000 to 20,000 g or greater at room temperature (e.g., 5 kg, 10 kg, 15 kg or greater), while maintaining a hardness of at least about 100 g, e.g., 500-1000 g at increased temperatures.

[0066] A cheese product may also be characterized by its melting property measured by an increase in a dimension of its shape when exposed to an increase in temperature. The increase may be an increase in a dimension such as, but not limited to, diameter, cross-section, or length. For example, the cheese product, when in the shape of a cylinder exhibits an increase in diameter at a temperature in the range of about 40°C to about 80°C, e.g., an increase in diameter of at least 50% or greater, e.g., 100%, 200% or greater.IV. Methods of Making Plant-based Cheese Products

[0067] The present disclosure provides methods of making the plant-based cheese products, which may be carried out, for example, using a Stephan Cooker, a Thermomix, or a Blendtec cooker. Such methods may include the steps of blending, cooking, cooling, zein dough formation and addition, pasteurization, and shredding / packaging for storage or shipping.(i) Formation of a Prolamin DoughA dough is a thick, malleable, sometimes elastic paste made by mixing flour with a smallamount of water or other liquid. A prolamin dough refers to a dough formed by the prolamin isolate with water, optionally combined with a further plasticizer. An acid plasticizer can be selected from citric acid, malic acid, tartaric acid, lactic acid, gluconic acid and / or acetic acid. The acid may be optionally combined with water to form an acid solution. The concentration of the acid in the solution may be at least 70% by weight, such as 80%, 85% or 88% by weight. Using a zein isolate as an example of a prolamin, a plasticized zein dough may be prepared by heating an acid in water solution to about 42°C to about 45°C. Then, the zein isolate is added to the solution and the mixture blended until a dough is formed. Finally, the zein dough is removed from the container, and any excess water discarded. One exemplary zein dough composition comprises 5% zein, 47.8% water, 0.4% Lactic acid. Zein may not absorb all the water or acid. Such formed zein dough is then combined with other components to form the plant-based cheese product. One exemplary plant-based cheese product comprises about 11 % zein dough.(ii) Combining, Blending and Mixing

[0068] After the prolamin dough is formed, the structural component(s) and fat(s) are combined with the prolamin dough and blended. In a representative but not limiting process, starch or a combination of starches and melted coconut oil are blended for 1 minute via any conventional means, then the prolamin dough is added to the mixture and blended in.(iii) Cooking, Heating and Thermal process

[0069] Cooking as described herein can be achieved by any means for heating mixtures as described herein, using any direct or indirect heating methods, such using any of various types of heating elements or ovens. The plant-based cheese product made by the present disclosure involves at least one cooking step, which occurs after blending the starch and oil mixture. The starch and oil are heated to at least 85°C, or in the range of about 85°C to about 90°C for a period of time ranging from about 3 - about 60 minutes, such as 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 12 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes, 45 minutes or 60 minutes, preferably while stirring. In one aspect, the starch and oil are heated to atemperature of about 85°C, about 86°C, about 87°C, about 88°C, about 89°C, or about 90°C. In one aspect, the starch and oil are heated to a temperature of 85°C, 86°C, 87°C, 88°C, 89°C, or 90°C.(iv) Cooling

[0070] Cooling is a process to reduce the temperature of food ingredients or mixtures. Cooling can be done by refrigeration, releasing hot steam, or bathing in a cold-water bath. The plant-based cheese product made by the present disclosure involves at least one cooling step. Such exemplary cooling occurs after the starch-oil blended mixture is cooked at 90°C for about 30 minutes. The cooling is done by releasing the hot steam in the mixture, and allowing the mixture temperature to drop to about 55°C. Another cooling step occurs after zein is incorporated into the starch mixture, which comprises refrigerating the post-zein addition mixture for about 48 hours, to allow the final formation of the plant-based cheese product.(v) Pasteurization

[0071] Pasteurization is to apply heat to reduce microbial presence and destroy pathogens in food ingredients or products. Most packaged food products require pasteurization to fulfill regulatory and safety requirements. For a plant-based packaged cheese product, it is required that all ingredients are sufficiently pasteurized, i.e., with adequate microbial destruction. But an extended high heat exposure may decrease prolamin’s functional characteristics. Balancing the two, a primal pasteurization condition is needed. The present disclosure encompasses various pasteurization conditions. These tested conditions include high temperature short duration (HTST), wherein the mixture is subject to 75°C for 3 minutes; low temperature long duration (LTLT), wherein the mixture is subject to 65°C for 30 minutes; and ultra-high temperature (UHT), wherein the mixture is subject to 90°C for 3 minutes, or for 30 minutes. Such pasteurization process is coupled with zein addition to the cooled starch-oil mixture, so the plant-based cheese products are formed. It is found HTST may be an optimal pasteurization condition. In one aspect, the pasteurization is followed by a cooking step, and is to reduce the temperature tobetween about 75°C to about 65°C. And the pasteurization may last for at least 3 minutes. In one aspect, the pasteurization is at temperature of about 75°C, about 74°C, about 73°C, about 72°C, about 71 °C, about 70°C, about 69°C, about 68°C, about 67°C, about 66°C, about 65°C, In one aspect, the pasteurization is at temperature of 75°C, 74°C, 73°C, 72°C, 71 °C, 70°C, 69°C, 68°C, 67°C, 66°C, 65°C, In one aspect, the pasteurization lasts for about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 12 minutes, about 15 minutes, about 20 minutes, about 25 minutes, or about 30 minutes. In another aspect, the pasteurization lasts for 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 12 minutes, 15 minutes, 20 minutes, 25 minutes, or 30 minutes. In another aspect, the pasteurization is at about 65 °C for about 30 minutes. In another aspect, the pasteurization is at about 75 °C for about 3 minutes.IV. Shredding, Packaging, Storage and Shipping

[0072] The present disclosure can provide a plant-based cheese product in any physical form as desired. If a shredded cheese is desired, the post-refrigerated product can be shredded using conventional shredding techniques and tools. If a block or chunk shaped cheese is needed, the post-refrigerated product can be shaped and / or packaged in the desired shape or amount. It is recommended that the products formed are shipped under low temperature, such as below about 4°C.

[0073] Having described several aspects, it will be recognized by those skilled in the art that various modifications, alternative constructions, and equivalents may be used without departing from the spirit of the present inventive concept. Additionally, a number of well- known processes and elements have not been described in order to avoid unnecessarily obscuring the present inventive concept. Accordingly, this description should not be taken as limiting the scope of the present inventive concept.

[0074] Those skilled in the art will appreciate that the presently disclosed aspects teach by way of example and not by limitation. Therefore, the matter contained in this description or shown in the accompanying drawings should be interpreted as illustrative and not in a limiting sense. The following claims are intended to cover all generic andspecific features described herein, as well as all statements of the scope of the method and assemblies, which, as a matter of language, might be said to fall there between.EXAMPLES

[0075] The following examples are included to demonstrate preferred aspects of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventor to function well in the practice of the present disclosure, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific aspects which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the present disclosure.Example 1. Selection of Plant Prolamin Isolates

[0076] One aspect of the present disclosure is identifying a plant prolamin isolate that helps provide a plant-based cheese product with properties more comparable to those of dairy cheese than known, commercially available non-dairy cheese analogs. For this purpose, various prolamin isolates obtained from selected commercial sources were characterized and tested. All tested samples were isolated from maize, and thus zein is the predominant protein. Specifically, zein samples from Flo Chemical Corp under the name “FloZein Products” were obtained and labelled as “Commercial Zein” or “Commercial”. They were further subjected to carotenoid removal treatments, such as ethanol wash. Such treated zein samples were labeled as C1 , C2, C3, C5, C6, C7, C13, A, or B, and were tested for their carotenoid profiles. Following testing as described herein, it was determined that carotenoids in a prolamin isolate may produce undesirable color and flavor characteristics in the plant-based cheese product, such that a “decarotenoided” or “decolorized” zein is preferred. The carotenoid profiles of tested zein samples were summarized in Table 1A-1C. Further, the different zein samples were characterized by their physiochemical features, such as protein, fat and moisture content, as were the density, color and melt / stretchability of resulting plant-based, prolamin cheese made with each sample (Table 2). FIGs. 1-2 show the color differences amongthe zein samples and the plant-based cheese products made from them. FIG. 3A shows the color spectra of these zein samples. Bulk density and water activity varied greatly among the samples (FIGs. 3B-3C), but all were considered to fall within the acceptable limits. Color was identified as an important quality parameter. Stretch tests showed good reproducibility in all batches, and good melting properties were observed for all samples (FIG. 3D). The acceptability of samples was summarized in FIG. 3E. FIG. 3F provides the selection criteria for decolorized zein. Through these tests, a specific carotenoid-removed zein sample, referred to herein as “Target Zein C1” (interchangeably also called “Target”, “Target Zein” or “C1” in the present disclosure), was identified as the first choice for making plant-based cheese products of the present disclosure. The water holding capacity of C1 was found to be comparable with another zein samples (FIG. 3H), but C1 had better oil retention and surface hydrophobicity than the others (FIGs. 3I-3J).Table 1A: Carotenoid Profiles of Different Zein Samples without Carotenoid RemovalTable 1B: Carotenoid Profiles of Different Zein Samples Obtained by Carotenoid RemovalTable 1C: Carotenoid Profile Comparison Between Target Zein and Zein withoutCarotenoid RemovalTable 2: Zein Samples and PropertiesNOTE: pH provided is the pH of the plant-based cheese product made from such zein sample.Example 2. Zein Dough Formation with or without Plasticizer

[0077] The present disclosure relates to a plant-based cheese product made with nonanimal derived ingredients. Such non-animal ingredients may include but not limited to, gums (including carrageenan, or gum Arabic), starches (such as tapioca flour and modified food starches), coconut oil, water, lactic acid, salt, aldopentose sugars (D- xylose), plant prolamin isolate such as corn zein [preferably carotenoid removed),gluconic acid, water, and natural / artificial flavors. Such cheese product can be made in a Stephan Cooker, a Thermomix, or a Blendtec cooker. An indispensable step of making such plant-based cheese product of the present disclosure is to prepare a zein dough. Such zein dough may be optionally plasticized by using a food grade acid and water mixture as a plasticizer. To make a plasticized zein dough, the first step is to heat acid (water) solution to about 42-45°C. Then, add a plant prolamin isolate, such as Target Zein C1 , to the solution and mix on speed 2 of a Kitchen Aid until a dough is formed. Finally, remove the zein dough from the Kitchen Aid® mixer with a dough hook, and discard the excess water. If no acid is added in the heating step, the zein dough formed is not plasticized. Table 3 provides an exemplary composition and process of preparing a plasticized zein dough using lactic acid as the plasticizer.Table 3. Zein Dough PreparationExample 3. General Procedure of Making Plant-based Cheese Product

[0078] As shown in FIG. 4A, a plant-based cheese product may be made by the general steps of blending, thermal processing, cooling, mixing with zein dough, refrigerating and packaging / shredding. One or more pasteurization steps may be added in the process as discussed in Example 6. In the blending step, water, starch, oil, and hydrocolloids were blended together in a Thermomix (or a Stephan cooker, or a Blendtec cooker) to form a starch mixture. Thermal process was to cook such starch mixture to about 85°C to about95°C. It The cooling step was to cool such heated starch mixture to a lower temperature, such as about 40°C to about 60°C, preferably about 45°C to about 55°C. Then adding zein dough to the cooled starch mixture and mixing the two. At the end of the mixing step, zein was incorporated into the starch mixture. The thus obtained product was refrigerated at about 1 °C to 4°C for conditioning for at least 48 hours. At the end of refrigeration, a plant-based cheese product was formed, and ready to be shredded or packaged. FIG. 4B-4C further provided exemplary processes in a Thermomix and in a Stephan cooker, respectively. Tables 4A-4B provided the ingredients names and amounts used in these two exemplary processes. Specifically, gums, such as carrageenan or gum Arabic, were mixed with starches, such as tapioca flour and modified food starches. The starch mixture was placed into a Thermomix (or a Stephan cooker or a Blendtec cooker). Next, coconut oil was melted and added to the Thermomix and blended for one minute or until evenly distributed into the starch mixture. Then, water, lactic acid, and salt were combined and added to the Thermomix. The ingredients were blended for about one minute on low speed and then heated and held at temperature of about 90°C for about 30 minutes, mixing at medium speed. After about 30 minutes, the mixture was continuously stirred and cooled until the mixture fell to a temperature between 45-55°C. The zein dough, plasticized or unplasticized as prepared in Example 2, and food flavors were then added to the Thermomix for blending with the cooled starch mixture. When the zein dough was fully incorporated into the starch mixture, the product was removed from the Thermomix and placed into a mold (such as a plastic, a glass, or a silicon mold), covered, and conditioned for at least 48 hours in a refrigerator at about 1 °C to about 4°C. After refrigeration, the plant-based cheese product was set and may be shredded, cut, sliced, blocked, and packaged as a plant-based cheese product. FIG. 5A-5B provided color photographs of different plant-based cheese products made by the present disclosure. Panelists were asked to evaluate the flavor and aroma of raw shreds. Panelists reported the samples were a little more acid and with slightly higher corn notes than standard cheese but considered them as acceptable. The flavor intensities were not significantly changed when samples were melted. It is therefore concluded that the plant-based cheese products made from various maize zein (prolamin) isolates were acceptable to customers as cheese analogs.Table 4A: Formula and Production of a Plant-Based Cheese Product Using a ThermomixTable 4B: Formula and Production of a Plant-Based Cheese Product Using a 12LStephan CookerExample 4. Analysis Methods for Functional Performance of the Plant-Based Cheese Products

[0079] The following methods were used to characterize and analyze the functional performance of the plant-based cheese products made by the present disclosure. The first test is the Cheese Fork Stretch test, which is used to measure the stretchability of cheese and cheese analog products. Cheese was cut into a 5 mm thick / 39.5 mm diameter (block cheese) or weighed into 15 g portions (shredded cheese) and placed into a Pyrex dish or Pyrex Petri dish. The sample was covered with either aluminum foil or a glass Petri dish cover. The sample were heated in the oven at 450°F for 5 minutes or in the microwave for 15-45 seconds. The microwave oven used is TOSHIBA Microwave Oven, MODEL EM925A5A-SS, with input 120V~60Hz and 1350 W 2450 MHz. Samples were cooled for 2 minutes. Using a ring stand to hold a 2-foot ruler upright, a fork was slid horizontally under the cheese sample (0.5”) into the center of the cheese and slowly pulled upwards at an angle of 45 degrees. The stretch recording was made at the height of the cheese right before strand breakage. The process was repeated in triplicates.

[0080] The second test is the Cheese Schreiber Melt Test. It is used to measure the meltability of cheese and cheese analog products. Cheese was cut into 5 mm thick / 39.5 mm diameter circles (block) or 15g of shredded cheese was placed in the center of a PRYEX® Petri dish or PRYEX® dish. Using a caliper, 3 cross sections were measured of the cheese sample (3 measurements were then averaged to obtain a singular value). The sample was covered with tin foil or a glass Petri dish cover and placed in an oven at 450°F for 5 minutes or at 400°F for 10 minutes or in a microwave heating for 15-45 seconds. The sample was removed and allowed to cool for 15 minutes. The diameter of the melted sample was measured at three cross sections (3 measurements were then averaged to obtain a singular value). Meltability was calculated using the following formula: melted cheese diameter — raw cheese diameter Meltability = - - - - - 100 raw cheese diameter

[0081] The third test is the Prolamin Microwave stretch. The plant-based cheese product (15 g) was shredded using a cheese box grater (largest hole size). The cheese shreds are arranged with a ring mold (95 mm) in an even layer on a 6 in porcelain plate. The rind mold is then removed, and the plate is placed into a microwave for 20 seconds. The sample is removed and allowed to cool to 85°C, measured using an infrared thermometer. Once cooled, the sample is placed at the base of a ring stand paired with a 2-foot ruler and the fork is slid into the cheese sample and pulled from the center. Measurements are made at the point right before the final cheese strand breaks. Measurements are repeated in 5 replicates.

[0082] The fourth test is the Cheese Stretch Texture Analyzer. It is used to measure the force required to stretch the cheese or cheese analog. A sample of the plant-based cheese product (15-20 g) was loaded into the well of the texture analyzer and the metal ring is placed over the cheese or cheese analog sample (ensure the probe is aligned with the TPA). The sample was placed into the oven (400°F for 10 minutes). The sample was removed from the oven and placed into the texture analyzer. The program for this test was run and the cheese or plant-based cheese analog begins to stretch. Once the strands begin to break, the test was stopped, and data collected. The test is repeated in triplicates.

[0083] The fifth test is Cheese TPA (Texture Profile Analysis). Cheese TPA is used tomeasure the firmness of the cheese product in block form. Cheese or cheese analogs are cut into 1” x 1” x 1” cubes and refrigerated for 3 hours prior to testing. The cheese is loaded into the TPA and the program run. Data is collected and the test run in triplicates. The sixth method of analysis is the Pizza Oven Stretch Test. Preheat a commercial conveyer belt pizza oven to 500°F. Place a 8” round pre-made pizza crust on a pizza peel and add 45.2 g of tomato sauce and 85 g of shredded cheese or cheese analog (leave %” around the edge of the crust and distribute cheese evenly across the surface of the pizza). Transfer the pizza to the conveyer belt and allow pizza to cook for 4-5 minutes. Remove pizza from the belt using the pizza peel and allow to cool for 2 minutes. Using a ring stand to hold a 2-foot ruler upright, a fork is slid horizontally under the cheese sample (0.5”) into the center of the cheese and slowly pulled upwards at a 45° angle. The stretch recording is made at the height of the cheese right before strand breakage. The process is repeated in triplicate.Example 5. Use of Gluconic Acid Instead of Lactic Acid

[0084] To reduce the tartness of the prolamin-based cheese product, to mimic the tartness level of real dairy-based Monterey Jack cheese, gluconic acid was evaluated as a replacement acid for lactic acid in making the zein dough. Results demonstrated that replacing lactic acid with gluconic acid reduced the tartness of the plant-based cheese product without affecting the pH values of the product and did not impact its stretch performance.

[0085] Gluconic acid is a mild organic acid with a brown and clear appearance. It is mainly present naturally in plants, fruits, wine, and honey. Gluconic acid has a pKa value of 3.86, similar to that of lactic acid (pKa=3.8). Therefore, when used at similar concentration, the plant-based cheese product containing gluconic acid should have a similar pH value to that containing lactic acid. The gluconic acid was evaluated at 50:50 lactic acid to gluconic acid replacement and at 100% lactic acid to gluconic acid replacement, with products containing 100% lactic acid as controls. Evaluation of sensory and textural attributes was conducted in both fully flavored and unflavored plant-based cheese products. Replacing lactic acid with gluconic acid reduced the sensory perceptionof tartness in both flavored and unflavored products. A reduction of bitterness also was observed in the flavored sample. There was no significant change in textural attributes such as firmness, tackiness, and chewiness. The melt stretch values of the plant-based cheese products reduced from 17.3 inches to 13.6 inches when all lactic acid was replaced with gluconic acid. At 50:50 lactic acid to gluconic acid replacement, there was no significant impact on melt stretch. The change in melt stretch was likely caused by the different plasticizing effect of the different organic acids, which are known to affect the thermomechanical properties of the cheese product. The pH values of the plant-based cheese product and the acid solutions were evaluated. Replacing lactic acid with gluconic acid did not seem to have a significant impact on pH values of the product and the acid solution, as predicted. Overall, the results showed that lactic acid can be potentially replaced with gluconic acid to modulate the tartness of the plant-based cheese products without impacting the pH and textural attributes.Example 6. Impact of Zein Content & Processing on Sensory and Functional Performance of the Plant-Based Cheese Products

[0086] Melt and / or stretch properties of the plant-based cheese product may be impacted by the zein incorporation process and / or by the amount used. Before the present disclosure, a minimum of 10% zein by weight was believed necessary to provide the stretch and melt properties desired for a cheese analog product. The present disclosure has demonstrated for the first time that using a zein isolate with carotenoids substantially removed, and using process modifications as shown in FIG. 4A, as little as 5% by weight of a zein isolate is sufficient to produce a plant-based cheese product with improved flavor, yet without significantly sacrificing melt and stretch properties.

[0087] Two alternative zein incorporation processes were tested. The first process involved plasticizing a zein isolate first before incorporating it into the starch mixture, as described in Example 2, to produce “pre-plasticized zein.” The second process incorporated a zein isolate in powder form directly into the starch mixture. FIG. 6A shows that the plant-based cheese made from pre-plasticized zein exhibited much better stretch than product made using zein powder. Direct incorporation of zein powder into the starchmixture produced plant-based cheese products having a low melt stretch of 3.4 inches. In comparison, use of the pre-plasticized zein dough produced a plant-based cheese product with a melt stretch of 7.1 inches, more than double the melt stretch of those made using zein powder. These results show that incorporation of pre-plasticized zein dough into the cooked starch mixture improves the melt stretch performance of the cheese product. Without wishing to be bound by theory, it is believed that pre-plasticizing the zein, for example with both acid and salt, results in the formation of a strong zein network, which improves the melt stretch performance of the final plant-based cheese product. In contrast, incorporation of zein powder directly into the product may hinder the aggregation of zein protein molecules to develop a strong network, thus resulting in lower melt stretch values. Additionally, different zein grades were tested by pre-plasticizing with acid then incorporating into the cooked starch mixture. Plant-based cheeses prepared by the different methods were compared with standard, dairy cheese products, such as a Commercial #1 cheese. FIG. 6B showed the plant-based cheese product made from Zein C1 showed the highest microwave stretch among all zein samples and stretch similar to Commercial #1 cheese.

[0088] Next tested was the impact of amount of zein on the stretch property of the plantbased cheese product. Plant-based cheese products were made by incorporating 5%, 6%, 7% and 8% of pre-plasticized C1 zein into cooked starch mixture. As shown in FIG. 7, the stretch distance of the cheese product increased as the zein content increased. At 5% zein content, the cheese product had a melt stretch of 7.8 inches, and at 8%, it increased to 15 inches. It was worth noting that increasing zein content would also increase the zein off-flavor notes in the cheese product made. In order to minimize zein off-notes, zein content was to be below 7%.Example 7. Thermal Processing and Pasteurization

[0089] Different pasteurization and thermal processing conditions were evaluated to fulfill regulatory and safety requirements on the plant-based cheese products. On one hand, it is required that all ingredients are sufficiently pasteurized, i.e., with adequate microbial destruction. On the other hand, extended high heat exposure may decrease zein's functional characteristics. Balancing the two, five different thermal processingconditions were tested in the Thermomix to determine the most optimal process to retain the zein functionality and fulfill the pasteurization requirements. FIG. 8A-8B showed the existing process with pasteurization at the cooking step, and the final plant-based cheese product was not pasteurized. FIG. 8C showed the five conditions tested with thermal processing at cooling and zein addition stages. The tested conditions were labeled as LTLT, HTST and UHT as defined in FIG. 8C.

[0090] Various functional tests were performed on the obtained plant-based cheese products. FIG. 9A showed the device / instruments used in these tests, such as Fork Stretch test, Schreiber Melt test, Texture Analyzer for Cheese Texture Profile Analysis (TPA), and Colorimeter for color analysis. Fork Stretch test measures stretchability, while observingstrand integrity, thickness and tenting. Schreiber Melt test measures meltability, while observing consistency and bubbling. Its rings help establishing the melt area. Sample preparation includes Texture Analyzer measures of force and resistance, it can calculate hardness, springiness, cohesiveness, and gumminess. Colorimeter measures color, and uses L* a* and b* values to show lightness / darkness, red / green, and yellow / blue. FIG. 9B provides the photographs of the five (5) plant-based cheese products obtained through the five (5) different thermal conditions. FIG. 9C provides the folk stretch results, showing that cheese products obtained through LTLT & HTST had comparable stretch height and variability to control, but UHT or heating at 90°C for 30 minboth noticeably decreased the stretch length of the cheese product made. FIG. 9D provides the results of Schreiber Melt tests. FIG. 9E provides the melt test results, showing LTLT, HTST, and UHT all resulted in cheese products with meltability comparable to control. FIG. 9F graphed the hardness of the cheese products, showing products’ hardness was not significantly impacted by the processing conditions. FIG. 9G-9I and Table 5 graphed color measurements, showing UHT processing created a noticeably darker yellow / red cheese color, while HTST, LTLT had insignificant color difference to the control, and thus were acceptable. FIG. 9J-9L graphed products’ cohesiveness, gumminess, and cheese sensory, showing cheese product made by HTST was the most similar to the control cheese in chewiness, flavor, and color. The results also showed that extreme pasteurization decreased creaminess and shred integrity while darkening the color of theplant-based cheese products.Table 5. Color Measurements and AcceptabilityAE represents perceived difference in distance between two samples’ L*, a*, and b* values.

[0091] In summary, for the stretch property, products made by LTLT (5.7) & HTST (5.4) had comparable average height and variability to control (6.3), while products made by UHT (4.0) and 90°C for 30 min (4.0) had noticeably less stretchiness. For meltability, products made by LTLT (38.8), HTST (35.5), and UHT (39.1 ) showed comparable melt to control (43). The negative control had almost 50% decreased melt (25.3). For texture, products’ hardness was not significantly impacted by the processing conditions and UHT had the greatest variability in measurements (SD = 1133). For color, UHT processing created a noticeably darker yellow / red cheese color (AE = 3.34), while products made by HTST (1 .65), LTLT (1 .74) and 90°C for 30 min (1 .38) had insignificant color difference to control. For sensory, extreme pasteurization decreased creaminess & shred integrity but darkened the color of the plant-based cheese products. Overall, plant-based cheese products made by HTST condition was the most similar to the control cheese in terms of chewiness, flavor, and color. FIG. 9M-9N further illustrated these results. In conclusion, HTST thermal condition was the most favorable as evidenced by acceptable functionality and time efficient. Further tests on scale up to pilot scale are to be performed, in order to verify product consistency between HTST condition and control against batch size, shear rate, and heating methods / distribution.Example 8. Pasteurization Coordinated with Zein Addition

[0092] In furtherance of the above tests on pasteurization conditions, zein addition wascoordinated with different pasteurization conditions as shown in FIGs. 10. The obtained plant-based cheese products were evaluated for color, sensory notes, melt and stretching properties. The study showed that thermal treatment darkened the color of the cheese products (FIG. 11A-11 B). For shred structure, the difference between LTLT, HTST and the control were minor. However, zein with starches were brittle, and resulted in particulate formation (FIG. 11 B). For flavor and aroma on the produced cheese shreds, panelists reported the shreds made by HTST and zein with starches tasted a little more acidic and with slightly higher corn notes than the other two. However, the flavor intensities were considered as acceptable in HTST made cheese products. For melted cheese shreds, panelists reported that corn flavor and aromatics were stronger in the cheese made from zein with starches, but not noticeable in the other pasteurizing conditions, when comparing to the control. In conclusion, LTLT or HTST was recommended in order to prevent excessive degradation of zein, and with a good balance between color change and sensory perception.

[0093] For melting capacities, it was noted that cheese made from zein cooked with starches had the biggest drop in melting capacity (20%) when comparing with that of the control (34%). The lowest drop occurred for cheese made with HTST pasteurization (25%, FIGs. 11C-11 D). For stretch, all products were similar to the control, except for zein mixed with starches whose stretch was about 3 inches less than the others (FIG. 11C). The above results were summarized in FIG. 11 E, showing zein with starches was not an acceptable option due to the unfavored color, flavor and functionality. Between LTST and HTST, HTST was considered better because it better reserved technical and sensorial properties of the plant-based cheese products. Therefore, HTST was the recommended pasteurization method to be further tested in scale-up trials.

[0094] Additional tests were performed on stretch and melting property (FIGs. 11 F- 11 H), and the results confirmed that LTLT and HTST had comparable stretch to the control and adding the zein at 90°C showed a decline in overall stretch. Melting capacity did not change substantially when pasteurizations were applied, with the exception for cooking at 90°C for 30 minutes which drastically decreased the melt spread of the produced plant-based cheese.Example 9. Discussion

[0095] The present disclosure showed when using plant prolamin isolate to make a plant-based cheese product, the prolamin content was proportional to the level of off- notes in the cheese product. It also influenced the stretch property. Contrary to the general perception that at least 10% by weight of zein is needed to make an acceptable plant-based cheese product, the present disclosure successfully demonstrates for the first time, a plant-based cheese product using as little as about 5% of a com prolamin isolate. The plant-based cheeses disclosed herein exhibit improved flavor, and comparable stretch and melt attributes relative to prolamin cheeses using 10% or more of plant prolamin. The present disclosure also demonstrates that not all prolamin isolates provide comparable plant-based cheese products in terms of properties such as melt, stretch, color and taste. As described herein, a com zein isolate with carotenoids removed is superior to those without carotenoids substantially removed. The present disclosure also provides improved processes for making a plant-based cheese product using a plant prolamin such as a zein. Water and / or a food grade organic acid can be used as the plasticizer for zein, and the acid may be chosen from citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid, and gluconic acid. The present disclosure demonstrates that gluconic acid, the least tart among these acids, performs similarly to lactic acid as a plasticizer in making the plant-based cheese products disclosed herein, except in spread features which are essential to some type of cheese products.

Claims

CLAIMSWe claim:

1. A plant-based cheese product comprising less than 10% by weight of a plant prolamin isolate, combined with a fat and a structural component in water, to yield a cheese product comprising a prolamin non-covalent network.

2. The plant-based cheese product of claim 1 , comprising no more than 4%, nor more than 5%, no more than 6%, no more than 7%, no more than 8%, or no more than 9% by weight of the plant prolamin isolate.

3. The plant-based cheese product of claim 1 , wherein the plant prolamin isolate is obtained from a corn.

4. The plant-based cheese product of any one of claims 1 -3, wherein the plant prolamin isolate comprises a zein or a functional analog thereof.

5. The plant-based cheese of claim 4, wherein the plant prolamin isolate comprises a corn zein.

6. The plant-based cheese of any one of claims 1-3, wherein the plant prolamin isolate is substantially free of carotenoids.

7. The plant-based cheese of any one of claims 1-3, wherein the plant prolamin isolate comprises about 75%, about 80%, about 81 %, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90% or about 91 % by weight prolamin protein, about 2% to about 20% by weight fat, and the balance by weight moisture.

8. The plant-based cheese of claim 7, wherein the prolamin protein is a zein,9. The plant-based cheese of claim 7, wherein the plant-based cheese product has a pH of 2.9 - 3.1 , 2.95 - 3.05, or 2.96 - 3.04.

10. The plant-based cheese product of claim 1 , comprising about 0.1 - 25% by weight of the fat, and 1 -30% by weight the structural component.11 . The plant-based cheese product of claim 1 , wherein the fat is selected from the group consisting of sunflower oil, canola oil, safflower oil, soybean oil, avocado oil,olive oil, corn oil, flaxseed oil, almond oil, coconut oil, peanut oil, pecan oil, cottonseed oil, palm oil, palm stearin, palm olein, palm kernel oil, rice bran oil, sesame oil, cocoa butter, shea butter, shea stearin, shea olein, palm kernel stearin, palm kernel olein, grape seed oil, hazelnut oil, Brazil nut oil, linseed oil, acai palm oil, passion fruit oil and any combination thereof12. The plant-based cheese product of claim 1 , wherein the fat comprises an oleic acid content of greater than 20% by weight.

13. The plant-based cheese product of claim 1 , wherein the structural component comprises a thickening agent and / or a gelling agent.

14. The plant-based cheese product of claim 1 , wherein the structural component is selected from the group consisting of a starch, a microbial or vegetable gum, a protein, a sugar polymer, and any combination thereof.

15. The plant-based cheese product of claim 1 , wherein the structural component is selected from the group consisting of arrowroot, cornstarch, katakuri starch, potato starch, sago, wheat flour, almond flour, tapioca, tapioca flour, starch derivatives, modified or pre-gelatinized starches, alginin, guar gum, locust bean gum, gellan gum, tara gum, gum Arabic, Konjac gum, xanthan gum, collagen, agar, carboxymethyl cellulose, pectin, carrageenan, and any combination thereof.

16. The plant-based cheese product of claim 1 , additionally comprising a plasticizer.

17. The plant-based cheese product of claim 16, wherein the plasticizer is selected from water, a food grade acid, glycerol, a sugar, and any combination thereof.

18. The plant-based cheese product of claim 17, wherein the food grade acid is selected from the group consisting of carboxylic acid, gluconic acid, citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid and any combination thereof.

19. The plant-based cheese product of claim 16, wherein the plasticizer comprises or consists of a combination of a food grade acid and water.

20. The plant-based cheese product of claim 19, wherein the food grade acid is lactic acid or gluconic acid.21 . The plant-based cheese product of claim 1 , comprising a plasticizer in an amount of up to 5% by weight.

22. A method of making a plant-based cheese product as defined in any one of claims 16-21 , comprising the steps of: i) combining the structural component with fat and water, and heating to a temperature of at least about 85 °C, thereby forming a mixture having a viscosity that permits formation of a non-covalent prolamin network at a temperature greater than the glass transition temperature of the plant prolamin isolate; ii) combining the plant prolamin isolate with the plasticizer to produce a plasticized plant prolamin composition; iii) at the temperature of (i), combining the mixture of (i) with the plasticized plant prolamin composition of (ii), to form a cheese mixture; iv) cooling the cheese mixture to a temperature between about 65°C to about 75°C for at least 3 minutes, thereby providing a pasteurized plant-based cheese product.

23. The method of claim 22, wherein heating in (i) is to a temperature of about 85 °C to about 95 °C.

24. The method of claims 22-23, wherein the plant prolamin isolate has a glass transition temperature of greater than 40°C.

25. The method of any one of claims 22 -24, wherein the cooling in (iv) is to about 65 °C for at least 30 minutes.

26. The method of any one of claims 22-25, wherein the plant prolamin isolate is a zein.

27. The method of any one of claims 22-26, wherein the zein is a corn zein.

28. The method of any one of claims 22-27, wherein the plant prolamin isolate is substantially free of carotenoids.

29. The method of any one of claims 22-28, wherein the plant prolamin isolate comprises about 75%, about 80% or about 90% by weight zein protein, about 2% to about 20% by weight fat, and the balance by weight moisture.

30. The method of any one of claims 22-29, wherein the plant-based cheese product has a pH of 2.9 - 3.1 , 2.95 - 3.05, or 2.96 - 3.04,31. The method of any one of claims 22-30, wherein the plasticizer is selected from water, a food grade acid, glycerol, a sugar, and any combination thereof.

32. The method of any one of claims 22-31 , wherein the food grade acid is selected from carboxylic acid, gluconic acid, citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid, and any combination thereof.

33. The method of any one of claims 22-32, wherein the plasticizer comprises or consists of a combination of a food grade acid and water.

34. The method of claim 33, wherein the food grade acid is lactic acid or gluconic acid.

35. A plant-based cheese product comprising water and a food grade acid in combination with less than 10% by weight of a com zein, wherein the corn zein is plasticized by the water and the food grade acid.

36. The plant-based cheese product of claim 35, wherein the food grade acid is selected from carboxylic acid, gluconic acid, citric acid, malic acid, tartaric acid, lactic acid, acetic acid, oxalic acid, and any combination thereof.

37. The plant-based cheese product of claim 35, wherein the food grade acid is lactic acid or gluconic acid.

38. The plant-based cheese product of any one of claims 35-37, which exhibits 3.5 to 7.5 inches of stretch when subjected to an industry standard Fork Stretch test.

39. The plant-based cheese product of any one of claims 35-38, which exhibits a melt spread of at least 25% to about 50% when subjected to an industry standard oven melt spread test.

40. The plant-based cheese product of any one of claims 35-39, which exhibits a AE of less than 3.0 as compared to a positive control sample.41 . The plant-based cheese product of any one of claims 35-40, wherein the corn zein is substantially free of carotenoids.

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