Refined protein component and refined protein powder

Abstract

The present invention relates to a refined protein component obtained from a plant source comprising: between about 70% and about 90% by weight of a protein; and between about 2% and about 7% by weight of calcium; wherein the refined protein component is derived from an alkaline extracted plant source; and wherein the refined protein component is insoluble in an aqueous solution having a pH of less than about 6. The invention also relates to refined protein powder.

Description

[0001] This application is a divisional application of the Chinese invention patent application with the application date of January 9, 2017, application number 201780015627.6, and invention name: "Product analogs or components of such analogs and preparation processes thereof".

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of priority to U.S. Provisional Application No. 62 / 276,030, entitled Dairy Product Analogs and Processes for Making Same, filed January 7, 2016, and U.S. Provisional Application No. 62 / 326,403, entitled Dairy Product Analogs and Processes for Making Same, filed April 22, 2016. All of the above-related applications are incorporated herein by reference in their entirety. In addition, other references or publications cited in this disclosure are also incorporated herein by reference in their entirety. Field of the Invention

[0004] The present disclosure relates to food products that are substantially or entirely derived from non-animal sources, but have color, flavor, nutritional content, and / or other qualities similar to dairy products and / or other foods. The present disclosure also provides a process for producing such dairy-like foods and / or other similar foods. Background of the Invention

[0006] Vegetarian and vegan diets offer many benefits to individual consumers and to the world's food needs. Such benefits include healthy nutrition (e.g., lower saturated fat, no cholesterol), no ethical or religious dietary conflicts, fewer negative environmental impacts (e.g., fewer greenhouse gases produced in production), and more efficient use of resources (e.g., less water used in production).

[0007] Dairy-like food products and other types of similar foods derived from soy, coconut and almonds are readily available to consumers. The demand for these vegetarian / vegan alternatives to dairy products has been exacerbated by consumer intolerance to certain dairy ingredients. However, the acceptance of milk substitutes and other similar food substitutes has been relatively low. This may be due to a small extent to the increased intolerance experienced by consumers to the ingredients of these foods (e.g., allergy to soy protein). To a large extent, the appearance, flavor and / or mouthfeel of commercially available milk substitutes and other similar food substitutes are not attractive to consumers. In addition, many commercially available milk substitutes do not contain the same protein and fat content as dairy products and therefore have lower nutritional value.

[0008] Therefore, the production of milk substitutes and other similar food substitutes from other natural sources is needed. However, many natural sources contain compounds that limit their application in food. For example, peas and other leguminous plants contain compounds that produce significantly different colors and / or flavors, which are unpleasant and difficult to mask with colorants and / or flavorings. These compounds may also affect the structure or stability of proteins and therefore affect the appearance, sensory and nutritional quality and shelf life of foods derived from such other natural sources.

[0009] Therefore, there is an unmet need for non-animal derived foods that have the color, taste, nutritional content and other qualities of dairy products and other similar food substitutes, do not challenge common nutritional sensitivities, and provide the consumer experience that consumers are accustomed to from dairy products and other similar food substitutes. The present disclosure provides such and related foods and refined protein components (protein components) or refined protein isolates (protein isolates, PI), and their production processes. The present disclosure is directed to solving these and other problems disclosed herein. The present disclosure is also directed to overcoming and / or improving at least one of the shortcomings of the prior art, which will become apparent from the discussion herein. The present disclosure is also directed to pointing out one or more advantages of using the products, methods and refined protein isolates and / or refined protein components disclosed herein. SUMMARY OF THE INVENTION

[0011] In addition to the embodiments discussed in this summary, other embodiments are disclosed in the specification, drawings, and claims. This summary is not meant to cover every embodiment; various combinations or variations are also contemplated by the disclosure.

[0012] Certain embodiments relate to refined protein isolates or refined protein fractions that can be used as an ingredient in various food products.

[0013] Certain embodiments relate to processes for producing a refined protein isolate or a refined protein fraction that can be used as an ingredient in various food products.

[0014] Certain embodiments are directed to food products comprising protein obtained substantially or entirely from non-animal natural and / or modified non-animal natural sources, but having color, flavor, nutritional content, and / or other qualities similar to dairy products.

[0015] Certain embodiments are directed to analogous foods that contain protein obtained substantially or entirely from non-animal natural and / or modified non-animal natural sources, but have color, flavor, nutritional content and / or other qualities similar to those foods they are intended to replace.

[0016] In some such embodiments, the dairy product analogs are dairy milk analogs that contain at least about 1% by weight of refined protein obtained from one or more non-animal natural or modified non-animal natural sources and have neutral colors defined as an L* value of at least about 70, an a* value between about -1.5 and about +1.5, and a b* value between about -12 and about +12. Certain embodiments are directed to methods for producing such dairy milk analogs, comprising the steps of:

[0017] a) obtaining at least one lipid component;

[0018] b) obtaining at least one color-neutral refined protein component from a non-animal natural and / or modified non-animal natural source;

[0019] c) blending together the at least one lipid and the at least one neutral color refined protein component and a water component to produce a mixture; and

[0020] d) emulsifying at least a portion of the mixture to provide a dairy milk analog.

[0021] In some embodiments, the dairy analogs are dairy yoghurt analogs, and the method for producing such dairy yoghurt analogs may comprise the following steps:

[0022] a) obtaining at least one carbohydrate component;

[0023] b) obtaining at least one lipid component;

[0024] c) obtaining at least one neutral color refined protein component from a non-animal natural and / or modified non-animal natural source;

[0025] d) blending the at least one carbohydrate component, the at least one lipid component, and the at least one refined protein component with a water component to produce a mixture;

[0026] e) heating the mixture;

[0027] f) emulsifying the mixture to produce an emulsified mixture;

[0028] g) cooling the emulsified mixture;

[0029] h) adding a fermentation microorganism to the emulsified mixture to produce a fermentation mixture; and

[0030] i) incubating the fermentation mixture at an elevated temperature until the fermentation mixture solidifies and acidifies to provide a dairy yogurt analog.

[0031] Certain embodiments relate to a neutral color refined protein component obtained from a non-animal natural and / or modified non-animal natural source. Certain embodiments relate to a method for obtaining such a neutral color refined protein component, comprising the following steps:

[0032] a) obtaining a protein preparation from a non-animal natural and / or modified non-animal natural source;

[0033] b) washing the protein preparation at a wash pH;

[0034] c) extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0035] d) separating the aqueous protein solution from non-aqueous components;

[0036] e) adding salt;

[0037] f) precipitating protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0038] g) separating the protein precipitate from non-precipitated components; and

[0039] h) washing the protein precipitate to obtain a neutral-colored purified protein fraction.

[0040] Certain embodiments relate to a refined protein component obtained from a non-animal natural and / or modified non-animal natural source. Certain embodiments relate to a method for obtaining such a refined protein component, comprising the following steps:

[0041] a) obtaining a protein preparation from a non-animal natural and / or modified non-animal natural source;

[0042] b) washing the protein preparation at a wash pH;

[0043] c) extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0044] d) separating the aqueous protein solution from non-aqueous components;

[0045] e) adding salt;

[0046] f) precipitating protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0047] g) separating the protein precipitate from non-precipitated components; and

[0048] h) washing the protein precipitate to obtain a refined protein fraction.

[0049] The present invention includes the following embodiments:

[0050] 1. A method for obtaining a certain yield of a refined protein component from a non-animal natural and / or modified non-animal natural source, comprising the following steps:

[0051] a) obtaining a protein preparation from said non-animal natural or modified non-animal natural source;

[0052] b) washing the protein preparation at a wash pH;

[0053] c) extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0054] d) separating the aqueous protein solution from non-aqueous components;

[0055] e) adding salt;

[0056] f) precipitating protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0057] g) separating the protein precipitate from non-precipitated components; and

[0058] h) washing the protein precipitate to obtain a substantial yield of a refined protein component.

[0059] 2. The method of item 1, wherein the non-animal natural or modified non-animal natural source is a plant.

[0060] 3. The method according to item 1, wherein the plant is a leguminous plant.

[0061] 4. The method according to item 1, wherein the leguminous plant is pea.

[0062] 5. The method of item 1, wherein the wash pH is between about 3 and about 5.

[0063] 6. The method of item 1, wherein the extraction pH is between about 8 and about 9.

[0064] 7. The method of item 1, wherein the added salt is calcium dichloride at a concentration between about 50 mM and about 100 mM.

[0065] 8. The method of item 1, wherein the precipitation pH is between about 5 and about 6.

[0066] 9. The process of item 1, wherein the process further comprises the step of adding between about 0.05% and about 12% by weight of a polysaccharide to the aqueous protein solution.

[0067] 10. The process of item 1, wherein the yield is between about 50% and about 90%.

[0068] 11. A refined protein component, wherein the refined protein component is obtained from a non-animal natural and / or modified non-animal natural source by the method of one or more of claims 1 to 10.

[0069] 12. The refined protein component of item 11, wherein the refined protein component is a paste comprising between about 4% and about 25% by weight protein, between about 0.1% and about 1.5% by weight calcium, and between about 50% and about 90% by weight water.

[0070] 13. The refined protein component of item 11, wherein said refined protein component is a dry powder comprising between about 70% and about 90% by weight protein, and between about 2% and about 7% by weight calcium.

[0071] 14. The refined protein component of item 11, wherein the relative emulsifying activity of the refined protein component is between about 0.1 and about 2 relative to the emulsifying activity of bovine serum albumin.

[0072] 15. The refined protein fraction of item 11, wherein the refined protein fraction comprises at least about 80% of protein bands having a molecular weight of less than 200 kDa on a denaturing protein gel.

[0073] 16. The refined protein component according to one or more of items 1 to 15, wherein said refined protein component is of neutral color.

[0074] 17. The refined protein component according to one or more of items 1 to 15, wherein the refined protein component is not neutral in color. BRIEF DESCRIPTION OF THE DRAWINGS

[0076] Embodiments of the present disclosure are described, by way of example only, with reference to the accompanying drawings.

[0077] Figure 1A -B provides a schematic diagram of the refined protein separation process, according to certain embodiments.

[0078] Figure 2 Shown is protein (%) extracted from eight flour / protein isolate (PI) sources at increasing pH values, according to certain embodiments.

[0079] Figure 3 Shown are the percent yields (wt / wt) at the extraction, precipitation and bulk steps for eight sources tested, according to certain embodiments.

[0080] Figure 4 It is shown that for processed feedstocks, the percent protein (weight of protein / dry weight of sample) generally increases from the starting commercial flour / protein isolate to the final refined protein isolate, according to certain embodiments.

[0081] Figure 5 Fat levels are generally slightly enriched during the procedure, according to certain embodiments.

[0082] Figure 6 The carbohydrate mass percentage is shown as a function of process step (mass / dry weight), according to certain embodiments.

[0083] Figure 7 The starch mass percentage is shown as a function of process step (mass / dry weight), according to certain embodiments.

[0084] Figure 8 The calcium mass % is shown to increase during the process due to complexation of Ca ions with the refined protein, according to certain embodiments.

[0085] Fig. 9 The phosphorus content is shown to be slightly enriched in the refined protein, according to certain embodiments.

[0086] Fig.10 It is shown that sodium is generally removed or significantly reduced during the process, according to certain embodiments.

[0087] Fig.11 Potassium is shown to be generally removed or significantly reduced during the process, according to certain embodiments.

[0088] Fig.12 Ash content is shown, according to certain embodiments.

[0089] Fig.13 The total peak area of ​​the HPLC trace of the extract from the original flour versus the refined protein is shown, according to certain embodiments.

[0090] Fig.14A -H displays the Fig.13 Relevant HPLC chromatograms.

[0091] Fig.15 Shown are the total peak areas measured in samples of protein isolate and refined protein isolate after a refining process, according to certain embodiments.

[0092] Fig.16A -K displays the Fig.15 Relevant GC chromatograms.

[0093] Fig.17A -F shows a GC chromatogram comparing a commercial protein isolate sample to the same plant source after a round of processing from flour to protein, according to certain embodiments.

[0094] Fig.18A -C shows the difference in taste attributes between emulsions produced with refined protein or rinsed commercial pea protein (Roquette S85F), according to certain embodiments.

[0095] Fig.19 Shown is a comparison of the aftertaste between emulsions produced with commercial proteins and emulsions containing refined proteins prepared according to certain embodiments.

[0096] Fig. 20 Shown is a texture comparison between emulsions produced with commercial proteins and refined proteins prepared according to certain embodiments.

[0097] Fig.21A and Fig. 21B Shown is a comparison of the appearance of emulsions produced with commercial proteins and refined proteins prepared according to certain embodiments. DETAILED DESCRIPTION OF THE INVENTION

[0099] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0100] definition

[0101] As used herein, the term "analogous dairy product" refers to a food product that is simulated by a dairy product analog and that the dairy product analog can serve as a substitute. Examples of analogous dairy products include, but are not limited to, milk (e.g., whole milk, milk containing 2% (by weight) fat [2% milk], milk containing 1% (by weight) fat [1% milk], milk containing no fat [skim milk], milk containing calcium supplements [calcium-fortified milk]), cream (crème fraiche), clotted dairy milk, single cream, double cream, whipping cream, cultured dairy milk, kefir, powdered dairy milk, condensed dairy milk, Khoa (a whole milk powder in India), evaporated dairy milk, ricotta, infant formula (infant formula), ricotta cheese, yogurt, and so on. formulas), baked milk, butter, buttermilk, ghee, smen, anhydrous milk fat, cheese, curds, paneer, whey, cottage cheese, quark, cream cheese, fromage frais, yoghurt, ayran, lassi, leben, clabber, gelato, ice cream, ice milk, frozen custard, frozen yoghurt, villi, kajmak, filmjoelk, piima, veal, dulce de leche, leche), skim yogurt (Skyr) and curd desserts (Junket).

[0102] As used herein, the term "analogous food product" refers to a food product that is simulated by a food product analog and that the food product analog can serve as a substitute. Examples of analogous foods include dairy products, supplements (powders, pastes for athletes, health), nutrition shakes / drinks, energy bars, meat substitutes, eggs substitutes (as ingredients in baking, formulations or as end consumer products like scrambles), spreads (spicy or sweet), snack foods and / or salad dressings / condiments.

[0103] As used herein, the term "color-neutral refined protein component" refers to a protein preparation whose color is defined as an L* (lightness) value of at least about 70, an a* (red / green) value between about -20 and about 20, and a b* (yellow / blue) value between about -30 and about 30, as measured according to the methods disclosed in Example 6 or Example 8. However, as a preliminary note, many of the embodiments disclosed herein may or may not be color-neutral, as that term is used herein.

[0104] As used herein, the term "dairy milk" refers to a white liquid secreted by the mammary glands of female mammals. Dairy milk is composed of an emulsion in which fat is dispersed in an aqueous solution containing proteins (e.g., casein, albumin), carbohydrates, inorganic salts and other ingredients. Suitable mammals from which dairy milk can be obtained include, but are not limited to, cows, sheep, goats, bison, donkeys, horses, camels, yaks, buffaloes, humans and other mammals. Dairy milk obtained from cows typically contains about 3.5% fat (whole milk). The fat level can be reduced to a standardized level to obtain different grades of milk containing 0% to about 75% (weight) of fat present in whole milk, including but not limited to 2% milk (milk containing 2% (weight) of fat), 1% milk (milk containing 1% (weight) of fat) and skim milk (milk that does not contain fat).

[0105] As used herein, the terms "dairy product analog," "dairy milk analog," and "dairy yoghurt analog" refer to foods that are produced to have one or more of the following characteristics: substantially similar (or similar) color, flavor, nutritional content, and other qualities to an analogous dairy product, dairy milk, or dairy yoghurt, respectively; and that can be used as a substitute for an analogous dairy product, dairy milk, or dairy yoghurt, respectively, but that are made from natural sources and / or modified natural sources. These terms encompass foods that may have the same or substantially similar superior qualities as an analogous dairy food, but may not have the same or substantially similar inferior qualities as an analogous dairy food. Some non-limiting examples of inferior qualities are the presence of saturated fat, the amount of fat, the presence of cholesterol, and the presence of lactose.

[0106] As used herein, the term "food product analog" refers to foods that have been produced to have one or more of the following characteristics: substantially similar (or similar) color, taste, nutritional content and other qualities to similar foods; and can be used as substitutes for similar foods, but they are made from natural sources and / or modified natural sources. These terms cover foods that may have the same or substantially similar advantageous qualities as similar foods, but may not have the same or substantially similar inferior qualities as similar foods. Some non-limiting examples of inferior qualities are the presence of saturated fat, the amount of fat, the presence of cholesterol, and the presence of lactose. Exemplary food applications include, but are not limited to, refined protein as a supplement (supplement powder, slurry for athletes, health supplements), nutritional shakes / beverages, energy bars,

[0107] Meat substitutes, egg substitutes (as an ingredient in baking, recipe ingredients or as end-consumer products such as scrambled eggs), spreads (spicy or sweet and mild), snack foods and / or salad dressings / condiments.

[0108] As used herein, the term "modified natural source" refers to a natural source that has been altered (eg, mutated, genetically engineered) from its natural state.

[0109] As used herein, the term "non-animal natural source" refers to naturally occurring plants, algae, fungi and / or microorganisms.

[0110] As used herein, the term "nutritional profile" refers to the type and amount of ingredients contained in a food and / or a component of a food. For example, the nutritional profile of a dairy product can generally be defined by the content of its ingredients, including but not limited to total protein content, total lipid content, total carbohydrate content, total dietary fiber content, calcium content, sodium content, the type and amount of vitamins present, and combinations thereof.

[0111] As used herein, the term "meal" refers to the raw grain or grits of a cereal plant.

[0112] As used herein, the term "microbe" is an abbreviation of microorganism and refers to a unicellular organism. As used herein, the term includes all bacteria, all archaea, unicellular protista, unicellular animals, unicellular plants, unicellular fungi, unicellular algae, all protozoa, and all chromista.

[0113] As used herein, the term "protein preparation" refers to a preparation obtained from a natural source and / or modified natural source containing protein. The term includes protein isolates, protein concentrates, protein flours, soybean meal (meal) and / or combinations thereof.

[0114] As used herein, the term "protein concentrate" refers to a protein material obtained from a natural source and / or modified natural source and having at least a portion (or a substantial portion) of one or more of the following components removed: carbohydrates, ash, and other minor components. It typically contains at least about 30%, 40%, 50%, 60%, 70% or 80% by weight protein.

[0115] As used herein, the term "protein isolate" refers to a protein material obtained from a natural source and / or modified natural source and having at least a portion (or a substantial portion) of one or more of the following components removed: insoluble polysaccharides, soluble carbohydrates, ash, and other minor components. It typically has at least about 40%, 50%, 60%, 70% or 80% (by weight) protein.

[0116] As used herein, the terms “a”, “an” (the indefinite articles “a” and “an”), “the”, “said” (the definite article “the”) and similar referents refer to both the singular and the plural, unless expressly indicated otherwise herein or clearly contradicted by context.

[0117] As used herein, the term "about" refers to a value or range of values ​​greater than or less than 1 / 10 of the stated value, but is not intended to limit the value or range of values ​​to only this broader definition. For example, a value of "about 30%" means a value between 27% and 33%. Each value or range of values ​​prefixed with the term "about" is also intended to cover embodiments of the stated absolute value or range of values.

[0118] As used herein, the term "comprising" means "including". Various variations of the word "comprising" have correspondingly varying meanings. Thus, for example, a method "comprising" steps 'A' and 'B' may consist of only steps 'A' and 'B' or may include one or more additional steps (e.g., steps 'A', 'B', and 'C').

[0119] The subheadings used in the detailed description of the invention are only for the convenience of the reader and should not be used to limit the subject matter found in the full text of the disclosure or the claims. These subheadings should not be used to interpret the scope of the claims or claim limitations.

[0120] Dairy Product Analogs

[0121] Certain embodiments are directed to food products comprising refined protein obtained substantially or entirely from non-animal natural and / or modified non-animal natural sources, but having color, flavor, nutritional content, and / or other qualities similar to dairy products.

[0122] In some embodiments, the dairy analogs provided herein are analogs of dairy milk. In other embodiments, the dairy analogs are analogs of foods that are usually derived from dairy milk, including but not limited to cooking milk, sweetened condensed milk, flavored milk, yogurt, cheese, smoothies, milk shakes, coffee cream, infant formula, diet drinks, nutritional supplement drinks, clinical nutritional drinks, beverage powders and frozen desserts (e.g., ice cream, soft ice cream, frozen yogurt, sundae ice cream, pudding, whipped toppings) or combinations thereof. In some embodiments, the dairy analogs are mainly, substantially or completely composed of ingredients derived from non-animal natural sources. In alternative embodiments, the dairy analogs are composed of ingredients that are partially derived from animal sources but supplemented with ingredients derived from non-animal natural sources. The dairy analogs provided herein can be prepared for human or animal consumption, including domesticated animals (e.g., dogs, cats), farm animals (e.g., cattle, pigs, horses), and wild animals (e.g., non-domesticated carnivores). The dairy analogs can be used for various purposes, including but not limited to the feeding and delivery of active ingredients (e.g., vitamins, minerals, nutrients, therapies).

[0123] The dairy analogs provided herein have one or more of the advantages discussed in this disclosure. For example,

[0124] 1. The dairy analogue is not derived from animals or is not primarily derived from animals and can therefore be produced in a more sustainable manner;

[0125] 2. The dairy analogs can be produced from a wide variety of non-animal natural sources (e.g., legumes, grains, flax), thereby expanding their nutritional content and opening up the possibility of local sourcing of ingredients in areas where crops currently used to make dairy analogs cannot be grown;

[0126] 3. The dairy analogs provide nutritional qualities similar to or better than similar dairy products;

[0127] 4. The dairy analog provides similar or superior levels of protein, carbohydrates, vitamins, minerals, and / or other health benefits compared to dairy milk [e.g., whole milk, 2% milk, 1% milk, skim milk, or other dairy milk] (e.g., the dairy analog may contain equal or higher levels of protein, about 75% or lower levels of saturated fat, about 25% or lower levels of sugar, and / or equal or higher levels of calcium; or

[0128] The yogurt product analogs may contain about 50% or less protein levels, about 75% or less saturated fat levels, and / or greater than 25% or less sugars, and equal or greater levels of calcium compared to dairy yogurts [e.g., whole yogurt, 2% yogurt, 1% yogurt, nonfat yogurt, or other dairy yogurts]; and / or

[0129] 5. The dairy analogs are produced to have substantially lower levels and / or no certain ingredients that are less favorable to the general health of consumers (e.g., total sugars, saturated fat, total fat and / or cholesterol) or certain ingredients that are less favorable to the health of a small number of consumers (e.g.,

[0130] lactose).

[0131] One feature of the dairy product analogs provided herein is that they can have a color that is similar or substantially similar to the analogous dairy product. The color of a food product can be determined, for example, using a colorimeter or spectrophotometer capable of measuring light reflectance and L*a*b color space (see, for example, Jovanka et al., (2008) Color Changes of UHT Milk During Storage, Sensors 8(9):5961-5974; Zare et al. (2013) Probiotic Milk Supplementation with Pea Flour: Microbial and Physical Properties, Food and Bioprocess Technology 6(5):1321-1331; Sanz et al., (2008) Yogurt enrichment with functional asparagus fibre, Effect of fibre extraction method on rheological properties, colour, and sensory acceptance. European Food Research and Technology. Vol. 227(5):1515-1521; Brewer and Rankin (1998) Color of Nonfat Fluid Milk as Affected by Fermentation. Journal of Food Science. 63(1):178-180). The dairy analogs provided herein have a color defined as having an L* value of at least about 70, an a* value between about -1.5 and about +1.5, and a b* value between about -12 and about +12, as measured according to the method disclosed in Example 6. Alternatively, the color of the food product can be evaluated by a human sensory panel.

[0132] The dairy analogs provided herein may also have flavors similar to similar dairy products, and / or flavors similar to or superior to existing alternative dairy products (e.g., soy milk, almond milk). The flavor of a food product may be evaluated, for example, by a variety of different methods, including but not limited to blind tasting performed by a human inspector or a human sensory expert.

[0133] Compared with similar dairy products, the dairy analogs provided herein can include less, similar or greater amounts of protein. In some embodiments, the dairy analogs include by weight between about 0.01% and about 40%, between about 0.5% and about 30%, between about 1% and about 20%, between about 1% and about 10%, between about 1.5% and about 10%, between about 2% and about 8%, between about 1% and about 3%, between about 2% and about 5%, or between about 3% and about 7% of the protein obtained from non-animal natural and / or modified non-animal natural sources. In some embodiments, the dairy analogs include at least 0.01%, 0.5%, 1% 1.5%, 2%, 3%, 5%, 7%, 10%, 15%, 20%, 30% or 40% (weight) of protein obtained from non-animal natural and / or modified non-animal natural sources. In some embodiments, in the dairy analog, the ratio of protein to lipid is about 1:5, 1:4, 1:3, 1:

[0134] 2, 1:1, 2:1, 3:1, 4:1, 5:1 or 10:1. The protein content of a food can be determined by a variety of methods, including but not limited to AOAC International reference methods AOAC 990.03 and AOAC 992.15 and combustion analysis (ISO 14891:2008). In some embodiments, the dairy analog has an amino acid profile that is similar or substantially similar to the analog dairy product. In some embodiments, the amino acid profile of the dairy analog is at least 50%, 60%, 70%, 80%, 90% or 95% identical (by weight) to the amino acid profile of the analog dairy product in terms of the type of amino acids and the total amount of each amino acid. In some embodiments, the amino acid profile of the dairy analog is at least 50%, 60%, 70%, 80%, 90% or 95% identical by weight to the amino acid profile of a protein preparation obtained from a non-animal source used to prepare the dairy analog, in terms of the type of amino acids and the total amount of each amino acid. In some embodiments, the dairy analog has an amino acid profile similar to, for example, a legume or a mixture of legumes, grains, and oilseeds. In some embodiments, the amino acid profile of the dairy analog has at least 50%, 60%, 70%, 80%, 90% or 95% identical by weight to the amino acid profile of a legume or a mixture of legumes used to prepare the dairy analog, in terms of the type of amino acids and the total amount of each amino acid.

[0135] Dairy product analog provided herein can also include lipid.In some embodiments, described dairy product analog comprises by weight between 0% and about 10%, between about 0.5% and about 8%, between about 1% and about 7%, between about 21.5% and about 6%, between about 2% and about 5%, between about 2.5% and about 4%, between 0% and about 4% or between about 2% and about 4% from non-animal natural source, improve non-animal natural source or the lipid obtained from the mixture of such source.The lipid content of food can be determined by several methods, including but not limited to AOAC International reference method AOAC 954.02 (AOAC International reference method AOAC 954.02). Examples of suitable lipids include, but are not limited to, docosahexaenoic acid, eicosapentaenoic acid, conjugated fatty acids, eicosanoids, palmitic acid, glycolipids (e.g., cerebrosides, galactolipids, glyconitrolipids, lipopolysaccharides, gangliosides), membrane lipids (e.g., ceramides, sphingomyelins, bacterial terpenoids), glycerides, second messenger signaling lipids (e.g., diglycerides), triglycerides, prenol lipids, prostaglandins, glycolipids, oils (e.g., non-essential oils, essential oils, almond oil, aloe oil, bitter almond oil, avocado oil, kapok oil, calendula oil, canola oil, corn oil, cottonseed oil, evening primrose oil, grape oil, grapeseed oil, hazelnut oil, jojoba oil, flaxseed oil, macadamia oil Oil), natural oil, neem oil, non-hydrogenated oil, olive oil, palm oil, partially hydrogenated oil, peanut oil, rapeseed oil, sesame oil, soybean oil, sunflower oil, synthetic oil, vegetable oil), ω-fatty acids (e.g., arachidonic acid, ω-3-fatty acids, ω-6-fatty acids, ω-7-fatty acids, ω-9-fatty acids) and phospholipids (e.g., cardiolipin, ceramide phosphorylcholine, ceramide phosphorylethanolamine, glycerophospholipids, phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phospho-sphingolipids, phosphatidylserine) or a combination thereof. In certain embodiments, examples of suitable lipids may be selected from one or more of the following groups: sunflower oil, canola oil, soybean oil, palm oil, cocoa butter.

[0136] In some embodiments, the dairy analog provided herein comprises carbohydrates similar to, substantially similar to, or reduced in quantity to similar dairy products. The carbohydrate content of food can be determined by a variety of methods, including but not limited to high performance liquid chromatography. Examples of suitable carbohydrates include but are not limited to sucrose, glucose, fructose, mannose, steviosides, artificial sweeteners, or combinations thereof. In certain embodiments, examples of suitable carbohydrates may be selected from one or more of the following groups: sucrose, glucose, fructose. In some embodiments, the dairy analog comprises carbohydrates between about 0.5% and about 15%, between about 1% and about 10%, or between about 3% and about 8% by weight. In some embodiments, the dairy analog comprises at least 0.5%, 1%, 3%, 5%, 8%, 10%, or 15% (weight) carbohydrates. In some embodiments, the dairy analog contains about 30%, 40%, 50%, 60% or 70% less total carbohydrates by weight than in an equivalent size serving of plain yogurt or dairy milk, regardless of fat content. In some embodiments, the dairy analog does not contain lactose. In some embodiments, the dairy analog contains less than 5%, 3%, 1% or 0.5% lactose by weight. In some embodiments, the dairy analog contains sucrose.

[0137] The proteins, lipids and carbohydrates contained in the dairy analogs provided herein may be derived from non-animal natural sources, modified non-animal natural sources, multiple non-animal natural sources, multiple modified non-animal natural sources or combinations thereof. In some embodiments, at least some of the proteins, lipids and / or carbohydrates are not derived from non-animal natural sources or modified non-animal natural sources but are completely identical, substantially completely identical or similar to the proteins, lipids and / or carbohydrates present in the non-animal natural sources or modified non-animal natural sources. For example, the protein may be produced synthetically or biosynthetically but contains polypeptide molecules having amino acid sequences that are completely identical, substantially completely identical or similar to the polypeptide molecules present in non-animal natural sources. In some embodiments, at least some of the proteins, lipids and / or carbohydrates are not derived from non-animal natural sources or modified non-animal natural sources. The proteins, lipids or carbohydrates may be composed of molecules having completely identical structures or mixtures of molecules having at least 2 different structures. In some embodiments, at least 25%, 50%, 75%, 90%, 95%, 99% or 100% (by weight) of the proteins, lipids and / or carbohydrates are derived from plants. For example, the protein of the plant may be derived from leguminous plants. Examples of leguminous plants include, but are not limited to, alfalfa, lentils, beans, clover, peas, fava coceira, frijole bola roja, frijole negro, lespedeza, licorice, lupine, mesquite, carob, soybean, peanut, tamarind, wisteria, cinnamon, chickpea, garbanzo, fenugreek, green peas, yellow peas, snow peas, lima beans, fava beans, black beans, baby beans, or combinations thereof. As a further example, the carbohydrates may be derived from sugar cane, sugar beets, agave, corn, or stevia, or combinations thereof. As a further example, the lipids may be derived from plants containing oilseeds, such as soybeans, sunflowers, oil palms, rapeseed, linseed, or other oil crops such as corn.

[0138] In some embodiments, the dairy analog provided herein comprises calcium in a similar amount to similar dairy products. In some embodiments, the dairy analog comprises calcium in a similar amount to similar calcium-fortified dairy products. In some embodiments, the dairy analog comprises calcium in a similar amount to similar calcium-fortified dairy products. In some embodiments, the dairy analog comprises calcium between about 5 mg and about 3,000 mg, between about 50 mg and about 1,500 mg, between about 100 mg and about 1,300 mg, between about 200 mg and about 1,000 mg, or between about 300 mg and about 600 mg per cup (8 ounces, 224 g). In some embodiments, the dairy analog comprises at least 5 mg, 50 mg, 100 mg, 200 mg, 500 mg, 1000 mg, 1500 mg, or 2500 mg of calcium per 8 ounces or 224 g. The calcium content of food can be determined by a variety of methods, including but not limited to inductively coupled plasma (ICP) analysis.

[0139] Dairy products are sold in a variety of forms: homogenized, but also, for example, with reduced fat, different types of fat (e.g., unsaturated lipids), lack of lactose, lack of permeate, or with added vitamins and minerals. Dairy analogs provided herein can be formulated to match or substantially match one or more of the nutritional profiles of these different types of dairy products. In some embodiments, the dairy analogs can have a nutritional profile that is healthier than similar dairy products. In some embodiments, compared to an equal amount of whole milk, whole fat yogurt, or other products made from whole milk, the dairy analogs can have reduced levels of total fat, reduced levels of saturated fat, reduced levels of cholesterol, increased levels of vitamins or minerals, increased levels of protein, reduced levels of sugar, or are composed of other sugars or sweeteners instead of lactose, or a combination thereof.

[0140] In some embodiments, the ingredients of the dairy analogs provided herein are natural or substantially natural, so the dairy analogs may be attractive to consumers who prefer natural ingredients (i.e., ingredients that have not been genetically engineered or have a risk of contamination at any stage of the food chain, such as by genetically modified materials (e.g., by soy products or soy-based components); ingredients that utilize organic farming techniques). In some embodiments, the dairy analogs do not contain gluten, so they are attractive to consumers suffering from celiac disease or consumers who choose not to consume gluten. In some embodiments, the dairy analogs contain less than 1%, 0.5%, 0.1% or 0% (by weight) of gluten. In some embodiments, the dairy analogs do not contain nuts (e.g., peanuts) that can cause strong allergic reactions. In some embodiments, the dairy analogs contain less than 1%, 0.5%, 0.1% or 0% nuts. In some embodiments, the dairy analogs do not contain ingredients that consumers may consider unhealthy (e.g., cholesterol, trans fats, lecithin, phosphates, food coloring agents, artificial sweeteners, or excess sodium). In some embodiments, the dairy analog is substantially free of or completely free of ingredients containing cholesterol, trans fats, phosphates, food coloring agents, artificial sweeteners, excess sodium, or combinations thereof. In some embodiments, the dairy analog has a shelf life of 2, 3, 4, or 6 months (when processed to extend the shelf life and stored under refrigeration), or 3, 6, 9, or 12 months (when processed under sterile conditions and stored at room temperature).

[0141] Method for producing dairy analogs

[0142] Certain embodiments relate to methods for producing the dairy analogs.

[0143] In some embodiments, the method is for producing a dairy milk analog, and may comprise one or more of the following steps, in sequence or not:

[0144] a) obtaining at least one lipid component;

[0145] b) obtaining at least one neutral color refined protein component from a non-animal natural and / or modified non-animal natural source;

[0146] c) blending the at least one lipid component and the at least one neutral color refined protein component with a water component to produce a mixture; and

[0147] d) emulsifying at least a portion of the mixture to provide a dairy milk analog;

[0148] The amounts and ratios of the at least one lipid and at least one neutral color refined protein component are thereby selected to provide a desired nutritional profile.

[0149] In some embodiments, the method is for producing a dairy milk analog, and may comprise one or more of the following steps, in sequence or not:

[0150] a) obtaining at least one lipid component;

[0151] b) obtaining at least one refined protein component from a non-animal natural and / or modified non-animal natural source;

[0152] c) blending the at least one lipid component and the at least one refined protein component with a water component to produce a mixture; and

[0153] d) emulsifying at least a portion of the mixture to provide a dairy milk analog;

[0154] The amounts and ratios of the at least one lipid and at least one refined protein component are thereby selected to provide a desired nutritional profile.

[0155] Methods for obtaining the at least one lipid component are known in the art. In some embodiments, the at least one lipid component is obtained from non-animal natural and / or modified non-animal natural sources. Methods for obtaining the at least one neutral-colored refined protein component from non-animal natural and / or modified non-animal natural sources are provided herein. In some embodiments, the lipid component and / or protein component are obtained as a concentrated slurry. In some embodiments, the lipid component and / or protein component are obtained in solid form. In some embodiments, the neutral-colored refined protein component is mixed with one or more other proteins before mixing with the at least one lipid component.

[0156] The water phase can be an aqueous liquid, including but not limited to purified water, tap water, bottled water, deionized water, mineral water or a mixture thereof.

[0157] The lipid, protein and water components can be mixed in various orders. In some embodiments, these three components are mixed simultaneously. In other embodiments, the lipid component is mixed with the protein component before the water component is introduced into the mixture. In other embodiments, the protein component is mixed with the water component before the lipid component is introduced into the mixture. In other embodiments, the lipid component is mixed with the water component before the protein component is introduced into the mixture.

[0158] Mixing of the lipid, protein and water components can be accomplished using a variety of mixing devices, such as mechanical stirrers and / or high pressure jet machines. These components can also be stirred or mixed manually. Mixing can continue until each component is evenly distributed in the mixture in fact.

[0159] In some embodiments, a carbohydrate component is further added. A wide variety of ingredients can be used as a carbohydrate component, including but not limited to starch, white sugar, flour, edible fiber and combinations thereof. Examples of suitable starch include but are not limited to maltodextrin, inulin, fructooligosaccharides, pectin, carboxymethyl cellulose, guar gum, corn starch, oat starch, potato starch, rice starch, wheat starch or a combination thereof. Examples of suitable flour include but are not limited to amaranth flour, oat flour, quinoa flour, rice flour, rye flour, sorghum flour, soy flour, wheat flour, corn flour or a combination thereof. Examples of suitable edible fiber include but are not limited to barley gluten, carrot fiber, citrus fiber, corn bran, soluble dietary fiber, insoluble dietary fiber, oat gluten, pea fiber, rice bran, head husks, soy fiber, soy polysaccharide, wheat gluten, wood pulp cellulose or a combination thereof. In some embodiments, the carbohydrate component does not include lactose or does not substantially include lactose. The carbohydrate component may be present in the water component before mixing. Alternatively, the carbohydrate component is added to the lipid and / or protein component or to a lipid, protein and water mixture.

[0160] In some embodiments, one or more other ingredients are further added. In some such embodiments, the one or more other ingredients are added to the water component before mixing. In other embodiments, the one or more other ingredients are added to the lipid and / or protein component or to the lipid, protein and water mixture. In some embodiments, the one or more other ingredients include calcium.

[0161] Emulsification can occur without additional mechanical energy, or mechanical energy is required (e.g., vortexing, homogenization, stirring, ultrasonic treatment or other suitable mechanical activities). When emulsification is assisted by a lower amount of mechanical energy (e.g., stirring in a conventional mixer under moderate shear between about 100 rpm and about 1,000 rpm), the average droplet size of the resulting emulsion is typically larger (e.g., at least about 75% of the droplets have a diameter greater than about 25 um). When emulsification is assisted by a higher amount of mechanical energy (e.g., homogenization in a high pressure [e.g., between about 35 bar and about 650 bar] 1- or 2-stage homogenizer [e.g., between about 1,000 rpm and about 10,000 rpm], or microfluidization homogenization [between about 500 bar and about 2,000 bar]), the average droplet size of the resulting emulsion is typically smaller (e.g., at least about 75% of the droplets have a diameter less than about 10 um). Nanoemulsion can be obtained by homogenizing in a microfluidizer or other suitable equipment. In some applications, in order to obtain a higher lipid emulsion, the lipid component can be gradually added during mixing. In some applications, heating can help emulsification. In some embodiments, emulsification is carried out at a temperature higher than room temperature, higher than 30°C, 40°C, 50°C, 60°C, 70°C or 80°C, between about 90°C and about 120°C, between about 30°C and about 60°C, or between about 40°C and about 50°C. Cooling is usually performed after heating. Emulsification can be monitored by taking out a sample of the mixture and analyzing it by methods such as microscopy, light scattering and / or refractometry.

[0162] The emulsion may have droplets of various sizes. In some embodiments, the emulsion is a polydisperse emulsion (i.e., an emulsion containing droplets with a wide distribution of droplet sizes). In other embodiments, the emulsion is monodisperse (i.e., an emulsion containing droplets with a narrow distribution of droplet sizes). In some embodiments, the emulsion is a microemulsion (i.e., a thermodynamically stable system in which droplets are dispersed in a continuous phase). In other embodiments, the emulsion is a nanoemulsion (i.e., a metastable [or kinetically stable] dispersion of a liquid in different immiscible liquids with a droplet size range of 1 nm to 100 nm). In some embodiments, the average droplet size of the emulsion is less than about 1,000 nm, less than about 750 nm, less than about 500 nm, less than about 250 nm, less than about 100 nm, or less than about 50 nm, between about 100 nm and about 800 nm, or between about 100 nm and about 300 nm. In some embodiments, droplet size is reduced to the lipid content of reducing emulsion and dairy milk analog provided herein.During emulsification, the degree of emulsification can be realized by changing some parameters and therefore the final texture of the emulsion can be controlled to a certain degree.The example of such parameter includes but is not limited to the type and / or quantity of lipid component, the type and / or quantity of protein component, the type and / or quantity of optional emulsifier, the quantity of mechanical energy used during emulsification, centrifugal or filtering technique, the pH of water component, the temperature during mixing, the quantity or its combination of optional salt in water component.

[0163] In some embodiments, the dairy milk analog is sterilized or pasteurized. Sterilization can occur by UV irradiation, heating (e.g., steam sterilization, combustion, or dry heating) or chemical sterilization (e.g., exposure to ozone). In some embodiments, sterilization kills more than 95% of microorganisms. For pasteurization, the dairy milk analog is heated to a certain temperature (e.g., between about 280°F and about 306°F) and kept at such a temperature for a period of time (e.g., between about 1 second and about 10 seconds). Suitable pasteurization steps are known in the field of food manufacturing and can be carried out and / or continued for multiple time periods at various temperatures. Pasteurization can be high temperature short time (HTST), "extended shelf life" (ESL) processing, high pressure pasteurization (HPP), ultra pasteurization (UP), ultra high temperature (UHT) or a combination thereof. A controlled cooling system can be used to cool the dairy milk analog quickly. In some embodiments, the dairy milk analog undergoes vacuum cooling to remove volatiles and water vapor after pasteurization.

[0164] In certain embodiments, the source and / or quantity of the component are selected to provide the composition of the appropriate level without the need for additional supplementation. For example, the use of the low-fat natural source from which a lipid component or a protein component is isolated can facilitate the production of the dairy milk analog that reduces fat. Such a low-fat natural source includes but is not limited to a partially defatted natural source. Alternatively, components can be extracted from natural sources and conventional or unconventional methods such as centrifugation or membrane separation methods can be used to remove lipids from the extract. Similarly, the careful selection of natural sources and the seasonal variation of the composition of the extracted dairy milk analog can provide other components of the appropriate level, including but not limited to vitamins, minerals, dietary fiber, antioxidants or a combination thereof. The level of the composition can be further adjusted by supplementing.

[0165] The dairy milk analog may optionally be dried to obtain a powder. Drying may be performed in any suitable manner, including but not limited to spray drying, dry mixing, coagulation, freeze drying, microwave drying, drying with ethanol, evaporation, refractory window dehydration or a combination thereof.

[0166] The dairy milk analogs provided herein can be used as a base for producing other dairy analogs having certain nutritional profiles. In some embodiments, such other dairy analogs are yogurt analogs. The method provided herein for producing yogurt analogs can include one or more of the following steps, in order or not:

[0167] a) obtaining at least one carbohydrate component;

[0168] b) obtaining at least one lipid component;

[0169] c) obtaining at least one neutral color refined protein component from a non-animal natural and / or modified non-animal natural source;

[0170] d) blending the at least one carbohydrate, the at least one lipid, and the at least one refined protein component with a water component to produce a mixture;

[0171] e) heating the mixture;

[0172] f) emulsifying at least a portion of the mixture to produce an emulsified mixture;

[0173] g) cooling the emulsified mixture;

[0174] h) adding a fermentation microorganism to the emulsified mixture to produce a fermentation mixture; and

[0175] i) incubating the fermentation mixture at an elevated temperature until the fermentation mixture solidifies and acidifies to provide a dairy yogurt analog;

[0176] The amounts and proportions of the at least one carbohydrate component, the at least one lipid component and the at least one refined protein component are thereby selected to provide a desired nutritional profile.

[0177] The dairy milk analogs provided herein can be used as a base for producing other dairy analogs having certain nutritional profiles. In some embodiments, such other dairy analogs are yogurt analogs. The method provided herein for producing yogurt analogs can include one or more of the following steps, in order or not:

[0178] a) obtaining at least one carbohydrate component;

[0179] b) obtaining at least one lipid component;

[0180] c) obtaining at least one refined protein component from a non-animal natural and / or modified non-animal natural source;

[0181] d) blending the at least one carbohydrate, the at least one lipid, and the at least one refined protein component with a water component to produce a mixture;

[0182] e) heating the mixture;

[0183] f) emulsifying at least a portion of the mixture to produce an emulsified mixture;

[0184] g) cooling the emulsified mixture;

[0185] h) adding a fermentation microorganism to the emulsified mixture to produce a fermentation mixture; and

[0186] i) incubating the fermentation mixture at an elevated temperature until the fermentation mixture solidifies and acidifies to provide a dairy yogurt analog;

[0187] The amounts and proportions of the at least one carbohydrate component, the at least one lipid component and the at least one refined protein component are thereby selected to provide a desired nutritional profile.

[0188] Examples of suitable fermenting microorganisms include, but are not limited to, Lactobacillus bulgaricus and Streptococcus thermophilus.The elevated temperature may be a temperature or temperature range suitable for the metabolic activity of the fermenting microorganism, for example, about 45°C.

[0189] In some embodiments, the dairy analogs provided herein can be combined with dairy products and / or other dairy analogs. For example, the dairy milk analogs provided herein can be combined with dairy milk, seed milk (e.g., flax seeds), nut milk (e.g., almond milk) or milk derived from cereals (e.g., rice milk).

[0190] Neutral color refined protein fractions, refined protein fractions and / or refined protein isolates

[0191] Certain embodiments relate to neutral colored refined protein fractions obtained from non-animal natural and / or modified non-animal natural sources. Certain embodiments relate to refined protein fractions or isolates obtained from non-animal natural and / or modified non-animal natural sources.

[0192] The exemplary embodiments disclosed herein can be used in a variety of food applications including, but not limited to, dairy products, refined protein as a supplement (supplement powder, slurry for athletes, health supplements), nutritional shakes / drinks, energy bars, meat substitutes, egg substitutes (as an ingredient in baking, a recipe raw material, or as an end-consumer product such as scrambled eggs), spreads (spicy or sweet and mild), snack foods, and / or salad dressings / condiments.

[0193] In some embodiments, the total protein content of the neutral color refined protein component is at least about 10%, between about 10% and about 70%, between about 20% and about 60%, between about 30% and about 50%, between about 10% and about 30%, between about 10% and about 20%, or between about 12% and about 16% by weight.

[0194] In some embodiments, the total protein content of the refined protein component and / or refined protein isolate is at least about 10%, between about 10% and about 70%, between about 20% and about 60%, between about 30% and about 50%, between about 10% and about 30%, between about 10% and about 20%, or between about 12% and about 16% by weight.

[0195] In some embodiments, the neutral colored refined protein component has a total bound calcium content of between about 0.1% and about 2%, between about 0.3% and about 1.7%, between about 0.5% and about 1.5%, or between about 0.5% and about 1% by weight.

[0196] In some embodiments, the refined protein component and / or refined protein isolate has a total bound calcium content of between about 0.1% and about 2%, between about 0.3% and about 1.7%, between about 0.5% and about 1.5%, or between about 0.5% and about 1% by weight.

[0197] In some embodiments, the neutral color refined protein component is a paste comprising between about 4% and about 25% (by weight) protein and between about 0.1 and about 1.5% (by weight) calcium and between about 50% and about 92% (by weight) water. In some embodiments, the neutral color refined protein component is a dry powder comprising between about 70% and about 90% (by weight) protein and between about 2% and about 7% (by weight) calcium.

[0198] In some embodiments, the refined protein component and / or refined protein isolate is a paste comprising between about 4% and about 25% (by weight) protein and between about 0.1 and about 1.5% (by weight) calcium and between about 50% and about 92% (by weight) water. In some embodiments, the neutral-colored refined protein component is a dry powder comprising between about 70% and about 90% (by weight) protein and between about 2% and about 7% (by weight) calcium.

[0199] An exemplary neutral-colored refined protein component has the following composition: at least about 80% of visible protein bands on a denaturing protein gel having a molecular weight of less than 200 kDa, at least about 80% of visible protein bands on a denaturing protein gel having a molecular weight of less than 150 kDa, and at least about 80% of visible protein bands on a denaturing protein gel having a molecular weight between about 50 kDa and about 100 kDa.

[0200] In some embodiments, the relative emulsifying activity of the neutral colored refined protein component is between about 0.1 and about 2 relative to the emulsifying activity of a bovine serum albumin (BSA), whey or rice protein control, as determined by the method disclosed in Example 7.

[0201] In some embodiments, the relative emulsifying activity of the refined protein component and / or refined protein isolate is between about 0.1 and about 2 relative to the emulsifying activity of bovine serum albumin (BSA), whey or rice protein controls, as determined by the method disclosed in Example 7.

[0202] In some embodiments, the neutral color refined protein component contains lower levels or fewer types of fatty acid oxidation decomposition products. Examples of fatty acid decomposition products include, but are not limited to, hexanal, nonanal, octanal, decanal, (E)-2-nonenal, (E,Z)-2,6-nonadienal, (E,E)-2,4-decadienal and / or γ-nonalactone.

[0203] In some embodiments, the refined protein fraction and / or refined protein isolate contains lower levels or types of fatty acid oxidation decomposition products. Examples of fatty acid decomposition products include, but are not limited to, hexanal, nonanal, octanal, decanal, (E)-2-nonenal, (E,Z)-2,6-nonadienal, (E,E)-2,4-decadienal, and / or γ-nonalactone.

[0204] In some embodiments, the neutral colored refined protein component has an emulsifying activity index of between about 25 m2 / g and about 100 m2 / g at pH 7, as determined by the method of Kinsella et al. (J. Agric. Food Chem., Vol. 26, No. 3, 1978).

[0205] In some embodiments, the emulsifying activity index of the refined protein component and / or refined protein isolate is between about 25 m2 / g and about 100 m2 / g at pH 7, as determined by the method of Kinsella et al. (J. Agric. Food Chem., Vol. 26, No. 3, 1978).

[0206] The neutral color refined protein component can be used in convenient amounts to enable the production of food products or to provide adequate protein fortification in food products. In liquid form, the neutral color refined protein component can be used directly to produce liquid food products. In powdered form, the neutral color refined protein component can be blended with dry food products and then optionally reconstituted by dissolving the food product in water; or added to other ingredients to produce baked products for human consumption; or used as a protein supplement.

[0207] The refined protein fraction and / or refined protein isolate can be used in convenient amounts to enable the production of food products or to provide adequate protein fortification in food products. In liquid form, the refined protein fraction and / or refined protein isolate can be used directly to produce liquid food products. In powdered form, the refined protein fraction and / or refined protein isolate can be blended with dry food products and then optionally reconstituted by dissolving the food product in water; or added to other ingredients to produce baked products for human consumption; or used as a protein supplement.

[0208] Certain embodiments relate to refined proteins (isolates and / or fractions) that may have one or more of the following characteristics:

[0209] Refined proteins containing between 5% to 97%, 20% to 90%, 30% to 85% or 40% to 80% by weight of protein obtained from one or more non-animal natural or modified non-animal natural sources. Refined proteins containing at least 5%, 10%, 20%, 30%, 40%, 50% or 60% by weight of protein obtained from one or more non-animal natural or modified non-animal natural sources.

[0210] wherein the color is defined as having an L* value between 60 and 90, an a* value between -6 and +6, and a b* value between -20 and +20, an L* value between 65 and 85, an a* value between about -4 and about +4, and a b* value between -18 and +18, an L* value of at least 65, an a* value of at least -5 to +5, and a b* value of at least -16 to +16; or a refined protein having an L* value of at least 80, an a* value of at least -3 and +3, and a b* value of at least -14 to +14.

[0211] In certain embodiments, the refined protein may be in the form of a paste, a wet suspension, or a dry powder.

[0212] In certain embodiments, the refined protein may have a dry solids percentage of at least 5%, 10%, 15%, 20%, 25%, or 30% by weight.

[0213] In certain embodiments, the ratio of calcium to protein in the refined protein may be between 0.5% w / w to 5% w / w, 1% w / w to 6% w / w, 3% w / w to 8% w / w, or 5% w / w to 10% w / w.

[0214] In certain embodiments, the refined protein may be neutral in color or non-neutral in color.

[0215] In certain embodiments, the pH of the refined protein may be between 4.5 and 11, 6.5 and 10, 5.5 and 8, or 5.7 to 6.7. In certain embodiments, the pH of the refined protein may be at least 5. In certain embodiments, the pH of the refined protein may be less than 9.

[0216] In certain embodiments, the moisture content of the refined protein may be between 3% and 90% by weight. In certain embodiments, the moisture content of the refined protein may be at least 4% by weight. In certain embodiments, the moisture content of the refined protein is less than 80% by weight.

[0217] In certain embodiments, the fat content of the refined protein may be between 1% and 30% by weight. In certain embodiments, the fat content of the refined protein may be at least 2% by weight. In certain embodiments, the fat content of the refined protein may be less than 25% by weight.

[0218] In certain embodiments, the carbohydrate content of the refined protein may be between 0% and 50% by weight. In certain embodiments, the carbohydrate content of the refined protein may be at least 0% by weight. In certain embodiments, the carbohydrate content of the refined protein may be less than 25% by weight.

[0219] In certain embodiments, the starch content of the refined protein is between 0% and 10% by weight. In certain embodiments, the starch content of the refined protein is at least 3% by weight. In certain embodiments, the starch content of the refined protein is less than 9% by weight.

[0220] In certain embodiments, the phosphorus content of the refined protein is between 0% and 6% by weight. In certain embodiments, the phosphorus content of the refined protein is at least 0.1% by weight. In certain embodiments, the phosphorus content of the refined protein is less than 4% by weight.

[0221] In certain embodiments, the refined protein has a sodium and / or potassium content of less than 0.5% by weight.

[0222] In certain embodiments, the ash content of the refined protein is between 0% and 20% by weight. In certain embodiments, the ash content of the refined protein is at least 1% by weight. In certain embodiments, the ash content of the refined protein is less than 10% by weight.

[0223] In certain embodiments, the reducing power of the refined protein is between 5% and 50%. In certain embodiments, the reducing power of the refined protein is at least 6%. In certain embodiments, the reducing power of the refined protein is less than 46%.

[0224] In certain embodiments, the total HPLC peak area of ​​the refined protein is between 20,000 and 250,000 for total extractable soluble sugars and organic acids. In certain embodiments, the refined protein has a total extractable soluble sugars and organic acids of at least 22,000. In certain embodiments, the refined protein has a total extractable soluble sugars and organic acids of less than 240,000.

[0225] In certain embodiments, the refined protein has a total peak area of ​​between 50,000 and 3,000,000 as measured by GC analysis of volatile compound components. In certain embodiments, the refined protein has less than 2,500,000 volatile compound components.

[0226] In certain embodiments, the refined protein has an isoflavone component of between 0% and 0.1% by dry mass. In certain embodiments, the refined protein has an isoflavone component of less than 0.075% by dry mass.

[0227] In certain embodiments, the refined protein has a tannin component of between 0% and 0.5% by dry mass. In certain embodiments, the refined protein has a tannin component of less than 0.3% by dry mass.

[0228] In certain embodiments, the instability index of the refined protein is between 0.2 and 0.6. In certain embodiments, the instability index of the refined protein is at least 0.22. In certain embodiments, the instability index of the refined protein is less than 0.57.

[0229] In certain embodiments, the refined protein is consistently produced in amounts of at least between 500 kg and 3000 kg, between 500 kg and 1000 kg, between 1000 kg and -2500 kg, and between 1000 kg and 3500 kg.

[0230] Method for obtaining a refined protein fraction and / or a refined protein isolate

[0231] Certain embodiments relate to methods for obtaining a neutral color refined protein component from a non-animal natural and / or modified non-animal natural source. Certain embodiments relate to methods for obtaining a refined protein component and / or a refined protein isolate from a non-animal natural and / or modified non-animal natural source.

[0232] Some advantages of the methods provided herein are that the methods can remove or substantially remove flavoring agents, fragrances, coloring agents, other agents or combinations thereof in the refined protein preparations, and thus make the refined protein preparations more suitable for use in food. The removal of such agents can also increase the shelf life of foods containing such neutral-colored refined protein components, refined protein components and / or refined protein isolates.

[0233] The methods provided herein for obtaining a neutral colored refined protein component from a non-animal natural and / or modified non-animal natural source may comprise one or more of the following steps, in sequence or not:

[0234] a. obtaining a protein preparation from a non-animal natural and / or modified non-animal natural source;

[0235] b. washing the protein preparation at a wash pH;

[0236] c. extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0237] d. separating the aqueous protein solution from the non-aqueous components;

[0238] e. Add salt;

[0239] f. precipitating the protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0240] g. separating the protein precipitate from non-precipitated components; and

[0241] h. Wash the protein precipitate to obtain a neutral-colored purified protein fraction.

[0242] The methods provided herein for obtaining the refined protein components and / or refined protein isolates from non-animal natural and / or modified non-animal natural sources may comprise one or more of the following steps, in sequence or not:

[0243] a. obtaining a protein preparation from a non-animal natural and / or modified non-animal natural source;

[0244] b. washing the protein preparation at a wash pH;

[0245] c. extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0246] d. separating the aqueous protein solution from the non-aqueous components;

[0247] e. Add salt;

[0248] f. precipitating the protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0249] g. separating the protein precipitate from non-precipitated components; and

[0250] h. Washing the protein precipitate to obtain a refined protein fraction.

[0251] The refined protein preparation obtained from a natural source may have various forms, including but not limited to protein concentrates, protein isolates, protein powders, protein meal; natural, denatured or reconstituted proteins; dried, spray-dried or undried proteins; enzyme-treated or untreated proteins; and mixtures thereof. It may consist of particles of one or more sizes, and may be pure or mixed with other components (e.g., other plant-derived components). The refined protein preparation may be derived from non-animal natural and / or modified non-animal natural sources, or from a variety of natural and / or modified natural sources. In some embodiments, the refined protein preparation is obtained from a plant. In some such embodiments, the plant is a leguminous plant. In some such embodiments, the leguminous plant is pea. The pea may be one of the components of a whole pea or pea, a standard pea (i.e., a non-transgenic pea), a commercial pea, a transgenic pea, or a combination thereof. In some embodiments, the pea is a vegetable pea (Pisum sativum). In some embodiments, the leguminous plant is a soybean. The soybean can be whole soybean or one of the components of soybean, standard soybean (i.e., non-transgenic soybean), commercial soybean, transgenic soybean, or a combination thereof. In some embodiments, the legume is chick pea. The chick pea can be whole chick pea or one of the components of chick pea, standard chick pea (i.e., non-transgenic chick pea), commercial chick pea, transgenic chick pea, or a combination thereof. In some embodiments, the refined protein preparation can be pretreated for various purposes, such as, for example, extracting the protein preparation in a solvent to remove lipids, and heating the protein preparation to remove volatiles.

[0252] Washing the purified protein preparation may be performed using a variety of methods including single washes, multiple washes, and / or countercurrent washes.

[0253] The washing pH and extraction pH can be suitable for washing and making the pH of protein solubilization in protein preparation. Suitable washing pH and extraction pH can be determined by the following method: test various pH conditions, and identify the pH conditions under which the best yield and quality of the refined protein component are obtained (by, for example, judging by one or more of the following characteristics of the protein component obtained: flavor, smell, color, nitrogen content, Ca content, heavy metal content, emulsifying activity, MW distribution and thermal properties). In some embodiments, the washing pH and extraction pH are alkaline pH. In some such embodiments, the alkaline pH is at least about 7.1, at least about 8, at least about 9, at least about 10, at least about 11, at least about 12, between about 7.1 and about 10, between about 8 and about 10, between about 9 and about 10 or between about 8 and about 9. In some such embodiments, the alkaline pH is about 8.5. In some embodiments, the washing pH and extraction pH are acidic pH. In some such embodiments, the acidic pH is less than 7, less than 6.95, less than 6.5, less than about 5, less than about 4, less than about 3, between about 2 and 6.95, between about 3 and about 6, or between about 3 and about 5. The extraction pH may be adjusted using a pH adjuster. In some embodiments, the pH adjuster is a food grade alkaline pH adjuster. In other embodiments, the pH adjuster is a food grade acidic pH adjuster. Examples of suitable acidic pH adjusters include, but are not limited to, acetic acid, hydrochloric acid, citric acid, succinic acid, and combinations thereof. Examples of suitable alkaline pH adjusters include, but are not limited to, potassium bicarbonate, sodium bicarbonate, sodium hydroxide, potassium hydroxide, calcium hydroxide, ethanolamine, calcium bicarbonate, calcium hydroxide, ferrous hydroxide, lime, calcium carbonate, trisodium phosphate, and combinations thereof. It can be used to obtain considerable extracted protein where feasible in order to provide an overall high product yield. The yield of protein in the protein aqueous solution may vary greatly, with typical yields ranging from about 1% to about 90%. The protein concentration of the aqueous protein solution is typically between about 1 g / L and about 300 g / L. The molecular weight distribution of the protein contained in the aqueous protein solution may vary widely.

[0254] The separation of the protein aqueous solution and the non-aqueous component can be accomplished by various methods, including but not limited to decanting the supernatant above the precipitate after centrifugation, or centrifuging in a sedimentation centrifuge. Centrifugation can then be followed by disc centrifugation and / or filtration (e.g., using activated carbon) to remove residual protein source materials and / or other impurities. The separation step can be performed at various temperatures in the range of about 1°C to about 100°C. For example, the separation step can be performed at a temperature between about 10°C and about 80°C, between about 15°C and about 70°C, between about 20°C and about 60°C, or between about 25°C and about 45°C. The non-aqueous component can be re-extracted with fresh solute at the extraction pH, and the protein obtained after clarification is mixed with the initial protein solution for further processing as described herein. Prior to further processing, the separated protein aqueous solution can be diluted or concentrated. Dilution is usually achieved with water, although other diluents can be used. Concentration can be achieved by a membrane-based method. In some embodiments, the diluted or concentrated aqueous protein solution comprises between about 1 g / L and about 300 g / L, between about 5 g / L and about 250 g / L, between about 10 g / L and about 200 g / L, between about 15 g / L and about 150 g / L, between about 20 g / L and about 100 g / L, or between about 30 g / L and about 70 g / L protein by weight.

[0255] The protein in the aqueous protein solution may optionally be concentrated and / or separated from soluble small molecules. Suitable methods for concentration include, but are not limited to, diafiltration or hydrocyclones. Suitable methods for separation of soluble small molecules include, but are not limited to, diafiltration.

[0256] Salt precipitation can be accomplished using various suitable salts and precipitation pH. Suitable salts, salt concentrations, polysaccharides, polysaccharide concentrations, and precipitation pH can be determined by testing various conditions and identifying the salts and pH and polysaccharide conditions (judged, for example, by one or more of the following characteristics of the obtained protein component) that obtain the most colorless and / or tasteless protein precipitates with the best yield and quality under these conditions. Flavor, smell, color, nitrogen content, Ca content, heavy metal content, emulsifying activity, MW distribution, and thermal properties. In some embodiments, salt precipitation occurs with calcium dichloride at a concentration between about 5mM and about 1,000mM. Other examples of suitable salts include, but are not limited to, other alkaline earth metal salts or divalent salts (e.g., magnesium chloride, sodium chloride, calcium permanganate, and calcium nitrate). Typically, the precipitation pH is opposite to the extraction pH (i.e., when the extraction pH is in the alkaline range, the precipitation pH is most suitable in the acidic range, and vice versa). In some embodiments, the precipitation pH is an acidic pH. In some such embodiments, the acidic pH is less than 7.1, less than about 6, less than about 5, less than about 4, less than about 3, less than about 2, between 6.9 and about 2, between about 6 and about 3, between about 6 and about 5, or between about 5 and about 4. In some such embodiments, the acidic pH is about 5.25. The precipitation pH can be adjusted using a pH adjusting agent. In some embodiments, the pH adjusting agent is a food grade acidic pH adjusting agent. In other embodiments, the pH adjusting agent is a food grade alkaline pH adjusting agent.

[0257] Separating the protein precipitate from non-precipitated components can be performed by one or more of the methods disclosed herein.

[0258] The washing of the protein precipitate can be performed by a variety of different methods. In some embodiments, the washing is performed at the precipitation pH.

[0259] The protein precipitate may optionally be suspended. In some embodiments, the suspension is at an extraction pH, for example, in the presence of a chelating agent (to remove calcium ions). If the suspended protein preparation is not transparent, it may be clarified by various convenient procedures (such as filtration or centrifugation).

[0260] The pH of the suspended neutral-colored refined protein component can be adjusted to a pH between about 1 and about 14, between about 2 and about 12, between about 4 and about 10, or between about 5 and about 7 by adding a food-grade alkaline pH adjuster (including, for example, sodium hydroxide) or a food-grade acidic pH adjuster (including, for example, hydrochloric acid or phosphoric acid).

[0261] The pH of the refined protein component and / or refined protein isolate may be adjusted to a pH between about 1 and about 14, between about 2 and about 12, between about 4 and about 10, or between about 5 and about 7 by adding a food grade alkaline pH adjuster (including, for example, sodium hydroxide) or a food grade acidic pH adjuster (including, for example, hydrochloric acid or phosphoric acid).

[0262] The neutral color refined protein fraction may be dried. Drying may be performed in any suitable manner, including but not limited to spray drying, dry mixing, coagulation, freeze drying, microwave drying, drying with ethanol, evaporation, refractory window dehydration, or a combination thereof.

[0263] The refined protein component and / or refined protein isolate may be dried. Drying may be performed in a suitable manner, including but not limited to spray drying, dry mixing, coagulation, freeze drying, microwave drying, drying with ethanol, evaporation, refractory window dehydration or a combination thereof.

[0264] Other optional steps in the methods provided herein are heating steps for the purpose of removing heat-labile contaminants and / or microbial contaminants; and additional filtering (e.g., carbon filtering) steps for the purpose of removing additional odors, fragrances and / or color compounds. In some embodiments, such additional filtering is performed immediately after the protein preparation is extracted or after the protein aqueous solution and the non-aqueous component are separated.

[0265] In some embodiments, the methods provided herein provide a yield (by weight) of protein of at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, between about 30% and about 90%, between about 40% and about 90%, between about 50% and about 90%, or between about 60% and about 90%.

[0266] Non-Animal Natural and Modified Non-Animal Natural Sources

[0267] The proteins, lipids, carbohydrates or other components of the dairy analogs provided herein can be derived from one or more non-animal natural and modified non-animal natural sources.

[0268] The proteins, lipids, carbohydrates or other ingredients of the food analogs provided herein can be derived from one or more non-animal natural and modified non-animal natural sources.

[0269] Suitable natural sources are naturally occurring plants, algae, fungi or microorganisms.

[0270] Examples of suitable plants include, but are not limited to, caraway, cilantro, cumin, fennel, parsley, dill, dandelion, helichrysum, marigold, wormwood, safflower, chamomile, lettuce, wormwood, marigold, lemongrass, sage, thyme, chia seeds, mustard, olives, coffee, peppers, eggplant, red peppers, cranberries, kiwis, vegetable plants (e.g., carrots, celery), marigolds, tansy, tarragon, sunflowers, , holly, basil, hyssop, lavender, lemon verbena, marjoram, lemon balm, patchouli, pennyroyal, peppermint, rosemary, sesame, spearmint, primrose, sambar, pepper, pimento, potato, sweet potato, tomato, blueberry, nightshade, petunia, morning glory glory), lilac, jasmine, honeysuckle, snapdragon, psyllium, elecampane, buckwheat, red amaranth, chard, quinoa, spinach, rhubarb, jojoba, cypselea, chlorella, marula, hazelnut, rapeseed, kale, bok choy, rutabaga, frankincense, myrrh, elemi, pumpkin, squash, curcurbit, manioc, dalbergia, legumes (e.g., alfalfa, lentils, beans, clover, peas, fava coceira, frijole bola roja, frijole negro), lespedeza, licorice, lupine, mesquite, carob, soybean, peanut, tamarind, wisteria, cinnamon, chickpea, chickpea, fenugreek, green pea, yellow pea, snow pea, yellow pea, lima bean, broad bean), geranium, flax, pomegranate, cotton, okra, neem, fig, mulberry, lilac, eucalyptus, tea tree, niaouli, fruit plants (e.g., apple, almond, peach, plum, pear, nectarine), strawberry, blackberry, raspberry, cherry, prune, rose, tangerine, lemon (e.g.,grapefruit, lemon, lime, orange, bitter orange, mandarin), mango, citrus bergamot, buchu, grapes, broccoli, brussels sprouts, camelina, cauliflower, rapeseed, rapeseed (canola), turnip, cabbage, cucumber, watermelon, honeydew melon, squash, birch, walnut, cassava, kapok, allspice, almond, breadfruit, sandalwood, macadamia, taro, tuberose, aloe, garlic, onion, shallot, vanilla, yucca, vetiver, galangal, barley, corn, curcuma aromatica), galangal, ginger, citronella, oats, palm, pineapple, rice, rye, sorghum, triticale, turmeric, yam, bamboo, barley, cajuput, canna, cardamom, maize, oats, wheat, cinnamon, sassafras, lindera benzoin, bay laurel, avocado, ylang-ylang, mace, nutmeg, moringa, horsetail, oregano, cilantro, chervil, chives, aggregate fruits, grain plants, herbal plants, leafy vegetables, non-grain legume plants, nut plants, succulents plants, land plants, water plants, delbergia, millet, drupes, schizocarps, flowering plants, non-flowering plants, cultivated plants, wild plants, trees, shrubs, flowers, grasses, herbs, brushes, lianas, cacti, green algae, tropical plants, subtropical plants, temperate plants, derivatives and hybrids thereof, or combinations thereof. In certain embodiments,Examples of suitable plants may be selected from one or more of the following: yellow pea, linseed, soybean, sunflower, rapeseed, sugar cane, sugar beet and corn.

[0271] Examples of suitable algae include, but are not limited to, viridiplantae, stramenopiles, rhodophyta, chlorophyta, PX, flordeophyceae, bangiophyceae, florideohpyceae, trebouxiophyceae, phaeophyceae, palmariales, gigartinales, bangiales, gigartinales, Chlorella, Laminaria japonica, Laminaria saccharina, Laminaria digitata, Macrocystis pyrifera, Alaria marginata, Ascophyllum nodosum, Ecklonia sp., Palmaria palmata, Gloiopeltis furcata, Porphyra columbina, Gigartinaskottsbergii, Gracilaria lichenoides, Chondrus crispus, Gigartina bursa-pastoris, derivatives and hybrids thereof, or combinations thereof. In certain embodiments, examples of suitable algae may be selected from one or more of the following algae: Schizochytrium sp., Chorella sp., Botrytis cinerea, Botryococcus braunii ) and Dunaliella tertiolecta.

[0272] Examples of suitable fungi include, but are not limited to, Candida etchellsii, Candida guilliermondii, Candida humilis, Candida utilis, Candida versatilis, Debaryomyces shansenii, Kluyveromyces lactis, Kluyveromyces marxianus, Kluyveromyces thermotolerans, Pichia pastoris, Rhodotorula sp., Saccharomyces bayanus, Saccharomyces beticus, Saccharomyces cerevisiae, Saccharomyces chevalieri, Saccharomyces diastaticus, Saccharomyces ellipsoideus, Saccharomyces exiguus, Saccharomyces florentinus, Saccharomyces pastorianus, Saccharomyces pombe, Saccharomyces sake, Saccharomyces suvarum, Sporidiobolus johnsonii, Sporidiobolus salmonicolor, Sporobolomyces roseus, Xanthophyllomyces dendrorhous, Yarrowia lipolytica, Zygosaccharomyces rouxii, derivatives and hybrids thereof, or combinations thereof.In certain embodiments, examples of suitable fungi may be selected from one or more of the following fungi: Saccharomyces sp., Pichia pastoris, Hansunula polymorpha, Aexula adeninivorans, Kluyveromyces lactis, Yarrowia lipolytica, and Schizosaccaromyces pombe.

[0273] Examples of suitable microorganisms include, but are not limited to, firmicutes, cyanobacteria (blue-green algae), bacilli, oscillatoriophcideae, bacillales, lactobacillales, oscillatoriales, bacillaceae, lactobacillus, arthrospira, Bacillus coagulans, Lactobacillus acidophilus, Lactobacillus Reuteri, Spirulina, Arthrospira platensis, Arthrospira maxima, derivatives and hybrids thereof, or combinations thereof. In certain embodiments, examples of suitable microorganisms may be selected from one or more of the following microorganisms: Escherichia coli, Lactobacillus sp., and Cornybacterium glutamicum.

[0274] Non-animal natural sources can be obtained from a wide variety of sources, including, but not limited to, nature (e.g., lakes, oceans, soil, rocks, gardens, forests, plants, animals), breweries, and commercial cell banks (e.g., ATCC, collaborative sources).

[0275] Improved non-animal natural sources can be obtained from a wide variety of sources, including but not limited to breweries and commercial cell banks (e.g., ATCC, collaborative sources), or can be produced from natural sources by methods known in the art (including selection, mutation or genetic manipulation). Selection generally includes continuous breeding and steady growth at multiple dilution rates under selection pressure. Mutation generally includes selection after exposure to mutagens. Genetic manipulation generally includes genetic engineering of target genes (e.g., gene splicing, insertion or deletion or modification by homologous recombination). Improved natural sources can produce non-natural proteins, carbohydrates, lipids or other compounds, or produce non-natural quantities of natural proteins, carbohydrates, lipids or other compounds. In some embodiments, the improved natural source expresses higher or lower levels of natural proteins or metabolic pathway compounds. In other such embodiments, the improved natural source expresses one or more new recombinant proteins, RNA or metabolic pathway components derived from another plant, algae, microorganism or fungus. In other embodiments, the improved natural source has an increased nutrient content compared to its natural state. In other embodiments, the improved natural source has more favorable growth and production characteristics compared to its natural state. In some such embodiments, the modified non-animal natural source has an increased specific growth rate compared to its natural state. In other such embodiments, the modified non-animal natural source can utilize a different carbon source than its natural state.

[0276] In some embodiments, the proteins, lipids, carbohydrates or other ingredients of the dairy analogs and / or food analogs provided herein are derived from byproducts of one or more non-animal natural or modified non-animal natural sources of previous processing. Examples of such byproducts include, but are not limited to, de-oiled meal (e.g., de-oiled linseed meal, de-oiled soybean meal, de-oiled sunflower meal, de-oiled rapeseed meal, or a combination thereof).

[0277] Other Ingredients

[0278] In some embodiments, the dairy analogs and / or food analogs provided herein contain additional ingredients that improve one or more of the following characteristics: color, taste, and nutritional and other qualities. Examples of such additional ingredients include, but are not limited to:

[0279] -Antioxidants (e.g., rosemary, spearmint, ascorbic acid, sodium ascorbate, Maillard browning products [melanoids], BHA, BHT, TBHQ, propyl gallate, tocopherol, vitamin A, vitamin E, carotenoids, flavonoids, or combinations thereof; between about 0.01% and about 10%, between about 0.05% and about 5%, or between about 0.1% and about 2% by weight);

[0280] - Sweeteners (e.g., glucose, sucrose, fructose, dextrose, maltose, dextrin, maltodextrin, sucralose, levulose, tagatose, galactose, natural sweeteners [e.g., agave, cane juice, corn syrup, honey, maple syrup, stevia or other compounds extracted from the stevia plant [e.g., rebiana-A, rebaudioside-A, reb-A]], sugar-free sweeteners [e.g., sugar alcohols such as maltitol, xylitol, sorbitol, erythritol, mannitol, isomalt, lactitol, hydrogenated starch hydrolysates], artificial sweeteners [acesulfame potassium K), aspartame, sucralose, saccharin, stevia, tagatose] or a combination thereof; the natural sweetener content may be at least about 0.01%, between about 0.1% and about 20%, between about 0.1% and about 10%, or between about 1% and about 6% (by weight); the artificial sweetener content may be at least about 0.01%, between about 0.05% and about 5%, or between about 0.1% and about 1.0% (by weight);

[0281] - vitamins (e.g., vitamin B12, vitamin D, vitamin C, vitamin A, vitamin E, vitamin B, vitamin K, thiamine, riboflavin, pyridoxine, carotenoids (e.g., beta-carotene, zeaxanthin, lutein, lycopene), niacin, folic acid, pantothenic acid, biotin, choline, inositol) or combinations thereof;

[0282] - emulsifiers (e.g., lecithin, carrageenan, cellulose gum, cellulose gel, starch, gum arabic, xanthan gum, mono- and di-glycerides, propylene glycol monoesters, sodium stearoyl-2-lactylate, polysorbate 60, polysorbate 80, lecithin, hydroxylated lecithin, or a combination thereof; the emulsifier content may be between about 0.01% and about 10%, between about 0.05% and about 5%, or between about 0.5% and about 2% (by weight);

[0283] - Stabilizers (e.g., starch, gums [e.g., xanthan gum, locust bean gum, gear gum, gum arabic, gum ghatti, gum karaya, gum tragacanth, gellan gum, gum], hydrated colloids [e.g., guar gum, gum arabic, locust bean gum, xanthan gum, gellan, carrageenan, cellulose, carboxymethyl cellulose, microcrystalline cellulose, methyl cellulose, hydroxypropyl methyl cellulose, hydroxypropyl cellulose, pectin, low methoxyl pectin, gelatin, agar, furcellaran, dextran, or a combination thereof; between about 0.1% and about 5%, between about 0.5% and about 3%, between about 0.7% and about 1.5% by weight; to enhance physical properties by imparting viscosity or mouthfeel properties, to stabilize and / or suspend insoluble materials, and to prevent separation or sedimentation of ingredients);

[0284] - preservatives (e.g., potassium sorbate, sorbic acid, or a combination thereof);

[0285] - buffering agents that prevent unwanted creaming or precipitation upon addition of the dairy milk analog to a hot, acidic environment (e.g., when added to hot beverages such as coffee; e.g., monophosphates, diphosphates, sodium carbonate and bicarbonate, potassium carbonate and bicarbonate, potassium phosphate, dipotassium hydrogen phosphate, potassium hydrogen phosphate, sodium bicarbonate, sodium citrate, sodium phosphate, disodium hydrogen phosphate, sodium hydrogen phosphate, sodium tripolyphosphate, or combinations thereof);

[0286] - Probiotics;

[0287] - Minerals (e.g., chloride, sodium, calcium, iron, chromium, copper, iodine, zinc, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, aluminum, soluble mineral salts, slightly soluble mineral salts, insoluble mineral salts, chelated minerals, mineral complexes, non-reactive minerals such as carbonate minerals, reduced minerals, ammonium, or combinations thereof);

[0288] -ω-3 fatty acids (e.g., docosahexaenoic acid [DHA]);

[0289] - antimicrobial agents;

[0290] -Sterols;

[0291] - dietary fiber (e.g., soy cotyledon fiber [e.g., in an amount between about 0% and about 40%, between about 1% and about 20%, or between about 1.5% and about 5% by weight];

[0292] - amino acids (e.g., essential amino acids [e.g., arginine, cysteine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, tyrosine, valine], amino acid salts, amino acid chelates, or combinations thereof);

[0293] - phospholipids;

[0294] - salt (e.g., sodium citrate, sodium chloride, potassium citrate, potassium phosphate, dipotassium phosphate, or a combination thereof; e.g., to enhance flavor and / or to provide a buffering effect to enhance protein stability);

[0295] - pH-adjusting agent (e.g., organic pH-adjusting agent, inorganic pH-adjusting agent, food-grade acid [e.g., acetic acid, lactic acid, hydrochloric acid, phosphoric acid, citric acid, tartaric acid, malic acid, gluconic acid, delta lactone (deltalactone), gluconic acid], alkaline pH-adjusting agent [e.g., disodium hydrogen phosphate, potassium hydroxide] or a combination thereof);

[0296] - a binder (e.g., carrageenan, cellulose gum, cellulose gel, starch, maltodextrin, gum arabic, xanthan gum, or a combination thereof; between about 0.01% and about 10%, between about 0.05% and about 5%, or between about 0.1% and about 2% by weight);

[0297] - prebiotics (e.g. fructooligosaccharides, galacto-oligosaccharides); and

[0298] - Biologicals (e.g., Bifidobacterium spp., Clostridium spp., Bacteroides spp., Enterococcus faecalis, E. coli, Enterobacter cloacae, Klebsiella pneumonia, Staphylococcus epidermidis, Staphylococcus haemolyticus, Lactoferrin, or a combination thereof).

[0299] The ingredients may be natural to one or more non-animal natural sources; may be produced from one or more modified non-animal natural sources; may be produced from one or more non-animal natural sources or modified non-animal natural sources under controlled conditions, or may be synthetically produced.

[0300] Packaging, labeling, marketing and sales

[0301] The dairy analogs and / or food analogs provided herein can be packaged in containers. Examples of suitable containers include, but are not limited to, bags, cups, jars, barrels, bottles, bowls, boxes, cans, cartons, bags-in-boxes, tubes, capsules, vacuum packaging, pouches, Tetrapak, bricks, gable tops, liquid aseptic packaging, roll-to-roll liquid aseptic packaging, and single-serving juice boxes, etc., and combinations thereof. The container can be heat-resistant and / or light-resistant. In some embodiments, the container is sterilized in advance. The packaged dairy analogs can be placed in refrigerated or frozen storage. In some embodiments, the flavor and texture of the dairy analogs are substantially maintained after being stored for at least 5 days. The packaging can carry one or more labels that convey information to consumers or support the marketing of food. Examples of information that may be communicated to consumers include, but are not limited to, no genetically modified organisms, no gluten, kosher, no cholesterol, vegetarian, vegan, no allergens, no soy, no nuts, or combinations thereof.

[0302] The dairy analogs and / or food analogs provided herein can be sold in a variety of suitable locations. Such locations include, but are not limited to, grocery stores, call centers, schools, vending machines, restaurants, sports venues, food service providers, and direct-to-consumer outlets.

[0303] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and / or were hereby incorporated in its entirety. Example

[0304] The following examples are included to demonstrate exemplary embodiments. It should be appreciated by those skilled in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventors of the present invention and that function well in the practice of the disclosed embodiments. However, it should be appreciated by those skilled in the art in light of the present disclosure that many changes may be made to the disclosed specific embodiments and still obtain the same or similar results without departing from the spirit and scope of the present disclosure. Therefore, the events shown or displayed in the accompanying drawings should be interpreted as illustrative, rather than limiting.

[0305] Example 1 - Obtaining a Neutral Color Refined Protein Fraction from Pea Protein Isolate

[0306] Spray dried pea protein isolate (Now Sports PeaProtein powder, Bloomingdale, IL) was added to distilled water and the pH was adjusted to pH 8.5 with 1N NaOH while stirring for 60 min to a final solid concentration of 100 g / L. The extract was separated by centrifugation at 5,000 to 15,000 g for 10 min. The supernatant was retained and the precipitate was discarded. The supernatant was centrifuged again to clear it from the solids. The pea protein was then precipitated by adding 500 mM CaCl2 to a final concentration of 50 mM and adjusting the pH to 5.25 with 1N HCl. The mixture was mixed briefly. When formed, the white precipitate was separated by centrifugation at 5,000 to 15,000 g for 10 min. The supernatant was discarded and the precipitate was washed twice with water (1 N HCl at pH 5.25) (acidified water was added to the precipitate to a final dilution of 25-50x (~1-2 mL of solid precipitate was mixed in 50 mL of acidified water; briefly mixed; centrifuged as above, repeated). The protein yields obtained were as follows: 7% (weight) extracted protein; 70% (volume recovery) protein aqueous solution; 33-50% mass yield (after salt precipitation); and 95% mass yield (after acid washing). The color of the resulting protein paste was determined as described in Example 6; the L* value of the protein paste was 89.5, the a* value was 4.1, and the b* value was 9.6, and the calcium to protein ratio was 6% w / w. The relative emulsifying activity (Table 1) of the protein paste was determined as described in Example 7.

[0307]

[0308] Example 2 - Obtaining a neutral coloured refined protein fraction from chick pea flour

[0309] Use hammer mill to grind dried chick pea to 30-100# mesh size to make chick pea powder. Add the chick pea powder to distilled water, adjust pH to pH 8.5 with 1N NaOH, and stir for 60min to final solid concentration of 100g / L. Separate the extract by centrifuging 10min at 5,000 to 15,000g. Keep the supernatant and discard the precipitate. The supernatant is filtered using tangential flow filtration (with a membrane of 100kDa molecular weight cutoff). Keep the retentate and discard the filtrate. By adding 500mM CaCl2 to a final concentration of 50mM, and adjust pH to 5.25 with 1N HCl, chick pea protein is precipitated. The mixture is mixed for a short time. When formed, separate the white precipitate by centrifuging 10min at 5,000 to 15,000g. The supernatant was discarded and the precipitate was washed twice with water (1 N HCl at pH 5.25) (acidified water was added to the precipitate to a final dilution of 25-50x (~1-2 mL of solid precipitate was mixed in 50 mL of acidified water; briefly mixed; centrifuged as above, repeated). The protein yields obtained were as follows: 70% (weight) extracted protein; 70% (volume recovery) protein aqueous solution; 33-50% mass yield (after salt precipitation); and 95% mass yield (after acid washing), and a calcium:protein ratio of 5% w / w.

[0310] Example 3A - Obtaining a Neutral Color Refined Protein Fraction from Pea Flour

[0311] Pea flour was made by grinding dried yellow peas (Giusto Specialty Foods, LLC) to 100# mesh size using a hammer mill. The pea flour was added to distilled water, adjusted to pH 8.5 with 1N NaOH, and stirred for 60 min to a final solid concentration of 100 g / L. The extract was separated by passing the slurry through a decanting centrifuge. The supernatant was retained and the precipitate was discarded. The supernatant was filtered using a tangential flow filtration (membrane with a 100 kDa molecular weight cutoff). The retentate was retained and the filtrate was discarded. The protein was precipitated by adding 500 mM CaCl2 to a final concentration of 50 mM and adjusting the pH to 5.25 with 1N HCl. The mixture was briefly mixed. When formed, a decanting centrifuge was used to separate the white precipitate. The color of the resulting protein paste was determined as described in Example 6; the L value of the protein paste was 81.9, the a value was 4.4, and the b value was 12.5, and the calcium:protein ratio was 7% w / w.

[0312] Example 3B - Obtaining Refined Protein from Pea Flour

[0313] Pea flour was made by grinding dried yellow peas (Giusto Specialty Foods, LLC) to 100# mesh size using a hammer mill. 10 kg of pea flour was added to distilled water, the pH was adjusted to pH 8.5 with 1N NaOH, and stirred for 60 min to a final solid concentration of 100 g / L. The extract was separated by passing the slurry through a decanting centrifuge. The supernatant was retained and the precipitate was discarded. The retentate was retained and the filtrate was discarded. The pea flour protein was then precipitated by adding CaCl2 to a final concentration of 10 mM and adjusting the pH to 4.5 with 6N HCl. The mixture was briefly mixed. When formed, a white precipitate was isolated using a disk stack centrifuge. The supernatant was discarded and the precipitate was washed with water (the water was titrated to pH 5.25 with 6N HCl). The yields obtained were as follows: 49% by weight of protein extracted from the pea flour, and 68% by weight of protein precipitated from the aqueous extract. This resulted in a final recovery of about 33% protein from the pea flour. The final mass of the refined protein isolate was 1.2 kg, containing 53% protein by mass (mass / dry weight %). The color of the emulsion produced from the protein paste was determined as described in Example 8; it had an L* value of 76.2, an a* value of -1.73, and a b* value of 1.2.

[0314] Example 4 - Production of Dairy Milk Analogs

[0315] A dairy milk analogue was produced, the composition of which is shown in Table 2.

[0316]

[0317] Cellulose gum and gellan gum are combined with evaporated cane juice in a dry blend. Sunflower oil is combined with DHA and lecithin to obtain an oil blend. Defoamer is added to water under high shear until dissolved, the neutral color refined protein component is added, and the mixture is mixed for 3 minutes at <1300rpm to obtain a liquid blend. Dipotassium hydrogen phosphate is added to the liquid blend, and the blend is mixed for another 1 minute. Dry blend, oil blend, mineral blend and flavoring are mixed together at >1,400rpm for 5 minutes to become a liquid blend. Record pH, and the blend is treated with microthermics for online homogenization (direct, 293F, 6.5 seconds, 2,500psi). The resulting product is collected and cooled, and then vitamins are added. The resulting dairy milk analog was determined as described in Example 6; the resulting dairy milk analog had an L* value of 75.9, an a* value of 0.2, and a b* value of 6.8; a protein content of 8 g per 8 oz serving; and a sugar content of 6 g per 8 oz serving. The taste was evaluated by a panel of panelists to see if it was superior to commercial almond milk and soy milk samples.

[0318] Example 5 - Production of Dairy Yogurt Analogs

[0319] A dairy yogurt analog base was produced, the composition of which is shown in Table 3.

[0320]

[0321] Combine corn starch / pectin blend with evaporated sucrose. Hydrate the neutral color refined protein component in 120°F water under high shear for 5 minutes. The pH of the hydrated neutral color refined protein component is adjusted to pH 7 with NaOH, then combined with corn starch / pectin / sugar blend under high shear and allowed to hydrate for another 30 minutes. Sunflower oil is added to the mixture under shear, and the mixture is heated to 140°F under constant stirring while covered with foil. The mixture is homogenized at 2,500psi (500psi Stage 2, 2,000psi Stage 1). Subsequently, the heating is increased to 200-205°F and maintained for 20min while maintaining stirring with a heat-resistant spatula. Finally, the mixture is covered with foil and left to cool to below 105°F. To 3,785.41 g of the dairy yogurt analog base, 1 g of Yogurtstarter Culture (Vivolac Soy 424) was added and the culture was incubated at 44° C. until pH 4.4-4.5 was reached. The resulting dairy yogurt analog had a low saturated fat content and no cholesterol compared to whole-fat yogurt, but a protein content similar to that of unfiltered whole-fat yogurt.

[0322] Example 6 - Method for determining the color of food

[0323] The color of the food was determined using a Datacolor 45S portable spectrophotometer (Datacolor, Lawrenceville, NJ, USA) using illuminant D65 and a viewing angle of 10 degrees. Calibration was performed using a reference tile and the results were expressed using the CIELAB system (determining L*-lightness, a* (green / red), and b* (blue / yellow)). The L*a*b values ​​for a number of dairy products were determined and are listed in Table 4. The color of food analogs may also be determined using this method or other acceptable methods.

[0324] Table 4 - L*a*b values ​​and protein content of commercially available dairy products and dairy analogs

[0325]

[0326]

[0327] Example 7 - Method for determining the relative emulsifying activity of a neutral colored refined protein fraction.

[0328] The neutral color purified protein fraction was dissolved in 0.3 M NaCl at 3.2% final protein concentration and 450 uL aliquots were added to each of three microcentrifuge tubes. A 0.3 M NaCl solution was used as a blank control, and a 0.3 M NaCl solution of bovine serum albumin (BSA; A2153, Sigma Aldrich) (3.2% final protein concentration) was used as a BSA standard. 50 uL of sunflower oil (Spectrum) was added to each microcentrifuge tube and vortexed for 15 minutes at setting 10 using a Genie Vortex and a foam tube adapter to produce an emulsion. Each tube was then placed on a stand and the emulsion was allowed to settle for 10-15 minutes. A 20uL aliquot of each emulsion (taken from the bottom of the tube, below any visible foam) was added to 480uL of 0.1% SDS solution, and the mixture was mixed by pipetting up and down 3 times (volume 300uL). Each sample was serially diluted 1:2 and 1:4 in 0.1% SDS. The absorbance at 500nm (A500) was measured for each sample as follows: using a 200uL aliquot of the sample, in a 96-well clear polystyrene plate, and using a Biotek Synergy H1 plate reader (programmed shaking for 1 minute to remove residual bubbles). The corrected A500 was calculated according to the following formula: Corrected A500 = (A500 sample - A500 blank) x dilution factor. The relative emulsification activity was calculated by dividing the average corrected A500 by the average corrected A500 of the BSA standard.

[0329] Example 8 - Process for isolating refined protein components and / or refined protein isolates from various flour and protein sources

[0330] The present embodiment relates to a process for producing a refined protein isolate or a refined protein component from a variety of plant protein sources. In the present embodiment, the analyzed plant sources include both leguminous plants and oilseeds. The present embodiment illustrates and quantifies the removal of various compounds (e.g., fat, carbohydrates, ions and / or small molecules) in the separation process, and evaluates the characteristics of the separated protein such as color, taste and functionality. In nine tested plant sources, the refined protein separation or refined protein component process is effective when separating clean refined proteins, and removes a considerable portion of carbohydrates, ions (except Ca) and small molecules. The process results in the production of white protein with a cleaner taste than currently comparable commercially available commercial proteins. The present embodiment is non-restrictive and the process, function and modification can be applied to one or more of other embodiments disclosed herein both qualitatively and quantitatively.

[0331] Plant Source

[0332] The plant sources tested included yellow pea (YPE), garbanzo bean (GAR), fava bean (FAV), white bean (WBF), kidney bean (NAV), soybean (SOY), sesame (SES), almond (ALM) and quinoa (QUI). Some commercial sources of protein used were: Pea Protein Isolate S85F (Roquette), source: Roquette; Puris Pea 870 (Puris), source: World Food Processing; Pea Protein Isolate 80 (Nutralliance), source: Nutralliance; and Soy Protein Isolate (Soy PI), source: Now Sports. The commercial sources of bean flours and oilseed flours used were Yellow pea flour (YPE) from Ingredion 1102; Garbanzo bean flour (GAR) from Bob's Red Mill; Fava bean flour (FAV) from Bob's Red Mill; White bean flour (WBF) from Bob's Red Mill; Navy bean flour (NAV) from whole navy beans from Bob's Red Mill, ground in a coffee grinder; Low fat soy flour (SOY) from Bob's Red Mill; Defatted sesame flour (SES) from Sukrin; and Defatted almond flour (ALM) from Sukrin.

[0333] Protein separation process

[0334] The refined protein separation process begins with a water extraction step, in which the plant source is mixed with water at a determined optimal pH for extraction. For extraction, preliminary tests for determining the optimal pH to solubilize plant proteins are described in the present disclosure, but other methods can also be used to determine the optimal pH. It should be understood that there may be one or more optimal pHs and / or one or more pH ranges that can be used in the embodiments disclosed herein. The aqueous phase is separated from the solids (starch / fiber) by centrifugation. The soluble protein is then precipitated by a combination of heating (60C), acid (pH reduced to 5.5) and CaCl2 addition (added to a ratio of 0.1g CaCl2 to g protein). The precipitated protein is separated by centrifugation and the supernatant is discarded. The separated protein is washed by mixing with tap water (30x dilution (weight)) and centrifuged again. The material from this final step is called refined protein isolate and is analyzed as described below. A schematic diagram of the process for flour type or protein isolate starting material is shown in Figure 1A . Figure 1A The steps used in the protein separation process for producing useful refined protein isolates and / or refined protein components for one or more of the embodiments disclosed below are schematically represented. Samples are prepared in duplicate according to the procedures described herein and certain process parameters (e.g., pH, temperature, and centrifugation speed) are kept constant or substantially constant; however, ranges of these parameters have been shown to be effective. In some embodiments, such as Figure 1B As outlined, the preliminary extraction step is bypassed.

[0335] In this example, an exemplary benchtop procedure (outlined in Figure 1B ) has the following steps:

[0336] 1.12% PPI loaded (weight) (90g PPI was brought to 750g with tap water)

[0337] 2. Mix on a mixing board until well combined.

[0338] 3. Raise pH to 9 with 6N NaOH and mix for about 5 minutes

[0339] 4. Add CaCl2 4M to reach a concentration of 30mM CaCl2

[0340] 5. Adjust pH to 4.6 with 6N HCl

[0341] 6. Centrifuge at 3500 rpm (2,200 g) for about 5 min. Discard the supernatant.

[0342] 7. Wash the precipitate with about 15x water (by weight).

[0343] 8. Centrifuge at 3500 rpm for about 5 min and discard the supernatant. The resulting precipitate is the final product.

[0344] In this example, the benchtop procedure for extracting refined protein isolate from flour consists of the following steps:

[0345] 1. If the alternative source is whole grains, grind them in a coffee grinder or pulverizer.

[0346] 2. 10% flour loaded (by weight, 250 g of flour and water added to bring it up to 2500 g).

[0347] 3. Mix on a mixing board until well combined.

[0348] 4. Adjust pH to 7 or 9 with 6N NaOH (use pH 7 for pulses and pH 9 for oilseeds, see Extraction pH Optimization).

[0349] 5. Centrifuge at 5000 rpm for about 5 minutes to separate. Collect the supernatant (extract). Discard the solid.

[0350] 6. If particles are still visible, pass the supernatant through another centrifugation (at 5000 rpm for about 5 min). If necessary, filter with a metal mesh.

[0351] 7. For sources with high fat content, the fat layer can be separated at this step. Pour the extract carefully to avoid the fat layer.

[0352] It should be understood that this embodiment is for bench preparation, and one or more steps and / or parameters can be changed to produce such useful refined protein isolates and / or refined protein components for larger or commercial quantities of useful refined protein isolates and / or refined protein components. The procedures set forth in this embodiment are exemplary, and other appropriate procedures can be used for both bench and commercial methods to obtain the refined protein isolates and / or refined protein components of the present disclosure.

[0353] Thereafter, the samples in this example were subjected to an exemplary CaCl protein precipitation process, comprising the following steps:

[0354] 1. Heat the extract in a water bath at about 63°C until the extraction temperature reaches 60°C.

[0355] 2. Based on the protein concentration, calculate the amount of CaCl2 4M that needs to be added to obtain a final ratio of 0.10 g CaCl2 / g protein.

[0356] 3. Add the corresponding volume of CaCl2 4M and adjust the pH to 5.5 (with 6N HCl) to precipitate the protein.

[0357] 4. Centrifuge at 5000 rpm (4650 g) for about 5 minutes.

[0358] 5. Pour off and discard the supernatant and collect the protein paste.

[0359] Thereafter, the sample in this embodiment undergoes an exemplary washing process, comprising the following steps:

[0360] 1. Resuspend the protein paste in 30x volume of water (by weight), mix with a rod homogenizer for 15 seconds, and adjust the pH to 5.5.

[0361] 2. Centrifuge at 4000 rpm (2970 g) for about 5 minutes.

[0362] 3. Pour off the supernatant and collect the protein paste. This is the final refined protein isolate. For each source, the optimization of protein extraction conditions

[0363] In this example, the extraction pH was optimized for each source. In general, for beans, pH 7 was a good compromise between good protein extraction yield and low extraction yield of other compounds (based at least in part on extraction color observations). For oilseeds, pH 9 had the greatest yield. The extraction pH used in the flour process at the time of the specific operation was 7 for beans and 9 for oilseeds. Figure 2 Shown is the protein (%) extracted from eight flour / protein isolate sources at increasing pH values.

[0364] Yield

[0365] This process was performed in duplicate on eight flour / protein isolate sources and the process described herein was used. This process was effective for the sources tested. The protein (%) solubilized and extracted from the flour (% extraction yield) varied in the range of 31-85% in the different sources, and the percentage of soluble protein precipitated from the extract (% precipitation yield) ranged from 21-71%. The overall yield from flour to protein isolate (% overall yield) ranged from 12-53%. For each source, the yield values ​​were similar in two replicates. The final percentage protein in the paste varied in the range of 7-20% (weight) depending on the plant source. Figure 3The % yield (by weight) in duplicate at each step is shown for the eight sources tested (extraction, precipitation, overall). Table 5 shows the results of the experiments for the protein isolation process using eight different plant sources. The yield at each step of the process, the protein extracted from the flour (extraction yield), the protein precipitated from the aqueous protein extract (precipitation yield) and the total yield from the process (overall yield). The amount of protein in the final mash varies depending on the plant source.

[0366] Table 5

[0367]

[0368] Refined Protein Macronutrients and Micronutrients

[0369] The following discussion shows that for this paradigm, the results of protein, fat, carbohydrate and starch content are functions of process steps. As discussed in the process description subsection, the starting raw material is a commercial flour / protein isolate made from various leguminous plants, oil seeds (defatted) or cereals. The commercial flour / protein isolate is then processed by water extraction, and the resulting supernatant is treated with CaCl2 and acid to solidify the insoluble portion, which is collected and analyzed. In general, the results are presented in the final recovered insoluble portion (exemplary refined protein isolate), with a considerable enrichment in protein content, a slight enrichment in fat, and a reduction in carbohydrates. Figure 4 It is shown that for the raw materials processed and tested, the % protein (protein weight / sample dry weight) generally increases from the starting commercial flour / protein isolate to the final protein isolate. This value is comparable to the protein value of commercial PI (measured by combustion using a LECO nitrogen analyzer). Figure 5 Fat levels generally showed a slight enrichment during the process, but it was noted that the fat to protein ratio decreased in the final protein isolate due to the increase in protein (fat was measured using AOAC 933.05). Figure 6 The carbohydrate mass percentage is shown as a function of the process step (mass / dry weight). In some cases, carbohydrates in the final product (final refined protein isolate) are undetectable. Oilseed defatted meal has lower carbohydrates than legume meal.

[0370] Figure 7The starch mass percentage is shown as a function of the process step (mass / dry weight). The starch reduction from the flour source (quinoa here instead of beans) is between 43% and 99% (from flour to final protein isolate). Once again, the oilseed meal has very low starch levels. With regard to the starch determination method used in this example, each sample was enzymatically digested (α-amylase (Sigma-Aldrich, AA306], amyloglucosidase (Sigma-Aldrich, A7095). Undigested samples were treated identically as controls. After incubation, the samples were vortexed and the supernatants were subjected to organic acid HPLC. The glucose in the undigested samples was subtracted from the glucose value in the digested samples. The net glucose concentration (weight) was back-calculated to find the original digested starch present in the sample. HPLC was performed on an Agilent 1200 HPLC instrument (using a 1260RID module). The analytical column was an Aminex 87-H column (Bio-rad, Hercules, CA] (equipped with a Micro-Guard Cation H cartridge (Bio-Rad)). The running buffer was 5 mM sulfuric acid (at a flow rate of 0.6 mL / min). The injection volume of the sample supernatant was 10 uL. The refractive index signal was observed for 30 minutes. For the analytes, the peak area was compared with an embedded five-point calibration (up to 30 g / kg).

[0371] Table 6 is a summary of the mass % (mass / dry weight) data of protein, carbohydrates, fat and starch for each replicate of each plant source tested:

[0372]

[0373] Ions (Ca, P, Na, K, Cl, ash)

[0374] Several ions that may impart flavor or nutritional content were measured during the process. Overall, the calcium content increased significantly during the process, which is by design. We coagulate the protein with calcium salts and the Ca++ ions become partially complexed with the protein and remain until the end of the process. Figure 8 The mass % calcium (mass / dry weight) showed an increase during the process, which was due to the complexation of Ca ions with proteins (measured using AOAC 984.27). There was a slight enrichment of phosphorus. Fig. 9 Phosphorus content was shown to be slightly enriched in the final refined protein isolate (measured using AOAC 984.27). Sodium and potassium ions were substantially washed out in the final refined protein isolate, which is at least partially due to the purification process. Fig.10 It was shown that sodium was generally removed or significantly reduced during the process (measured using AOAC 984.27). Fig.11It was shown that potassium was generally removed or significantly reduced during the process (measured by AOAC 984.27). Chloride ion was generally undetectable throughout the process (data not shown). Ash generally remained constant during the process (measured by AOAC 950.14A), such as Fig.12 Table 7 below provides a summary of the mass % (mass / dry weight) data of ions (Ca, P, Na, K) and ash for each replicate of each plant source tested:

[0375] Table 7

[0376]

[0377] Table 8 shows the percent change from commercial flour / protein isolate mass % (mass / dry weight) to refined protein isolate mass % (mass / dry weight) for protein, fat, carbohydrates and ions (Ca, P, Na, K) and is calculated based on the values ​​in Tables 6 and 7.

[0378] %change protein Fat carbohydrate calcium phosphorus sodium Potassium Almond Rep1 5 5 8 254 64 -89 -96 Almond Rep 2 27 25 -100 202 29 -93 -98 Sesame Rep1 82 -77 -100 81 -66 -93 -98 Sesame Rep2 66 -66 -100 5 -63 -90 -99 Soybean Rep 1 26 -14 -48 526 121 -92 -96 Soybean Rep 2 46 18 -100 512 109 -93 -97 Broad Beans Rep 1 163 33 -92 1760 158 -94 -98 Broad Bean Rep 2 175 26 -100 1615 196 -94 -98 Chickpea R1 207 70 -100 -78 62 -61 -98 Chickpea R2 170 79 -100 568 124 -89 -98 Kidney Bean R1 1 18 2 -92 654 219 -63 -95 Bean R2 123 18 -87 843 199 -66 -95 White Kidney Bean R1 247 211 -100 2256 763 -61 -89 White Kidney Bean R2 201 20 -94 449 21 1 -68 -94 Yellow peas R1 198 47 -84 1695 226 -69 -97 Yellow peas R2 171 56 -100 1023 141 -81 -98 Quinoa R1 36 -51 22 298 -82 -6 -93

[0379] Table 8

[0380] Restore capability

[0381] Reducing power is a proxy measurement of phenolic acids and other reducing compounds. Phenolic acids often contribute to the sour, bitter, and astringent tastes present in plant protein extracts; examples include flavonoids and tannins (see Huang et al., 1991; Roland et al., 2013). The overall reduction in reducing power was from 31% to 61% (from extract to final refined protein isolate). Table 9 below shows that reducing power was quantified using a modified Folin-Ciocalteu assay, reported as mg / L of gallic acid equivalents. The reducing power decreased by 31-86% from extract to refined protein isolate.

[0382] The reducing capacity of each sample was measured using a microplate adaptation of the Folin-Ciocalteu method, adapted from (et al., 2010 "Rapid microplate high-throughput methodology for assessment of Folin-Ciocalteu reducing capacity" Talanta 83; 441-447).

[0383] 1. Extract and purified protein isolate samples were diluted with water to achieve 0.3% protein by weight.

[0384] 2. The sample preparation in the microtiter plate consists of a mixture of the following substances (in this order):

[0385] a.50uL of sample / blank / standard

[0386] b. 50uL of Folin-Ciocalteu reagent 1:5 v / v (or 50uL 0.4M HCl for matrix blank)

[0387] c. 100uL of 0.35NaOH

[0388] 3. Incubate the samples at room temperature for 3 minutes.

[0389] 4. Prepare calibration samples using five gallic acid standards ranging from 5 mg / L to 150 mg / L (prepared daily from 500 mg / L gallic acid stock solution just prior to use).

[0390] 5. The absorbance of the samples was measured at 760 nm in triplicate. The reducing power was expressed as gallic acid equivalent (GAE).

[0391] 6. During assay development, a unit pulse of 50 mg / L GA was added to some samples.

[0392] Table 9.

[0393]

[0394] HPLC analysis was used to quantify the total extractable soluble sugars and organic acids in the original commercial flour isolate and the refined protein isolate to demonstrate the level of purification during the refining process. The samples analyzed on the HPLC were normalized to have the same number of proteins and the supernatant was injected onto an H-column (no acid digestion) after centrifugation. Although we did not identify the individual peaks in the HPLC traces, we compared the total peak areas of the flour and refined protein isolate. These chromatograms show a reduction in the total peak area of ​​the refined protein isolate - the total peak area was reduced by ∼99% in each sample. Fig.13 Shown is the total peak area of ​​the HPLC trace of the extract from the original flour versus the refined protein isolate; only ~1% of the original small molecules remain in the extract after the separation procedure (note the logarithmic scale). The actual chromatogram is shown in Fig.14A -G. Fig.14A The chromatogram is of defatted almond flour diluted to 6.9% protein. Fig. 14B is a chromatogram of a refined protein isolate from defatted almond meal (6.9% protein). Fig. 14C This is a chromatogram of bean flour ground in house and diluted to 3.6% protein. Fig.14D is a chromatogram of a refined protein isolate from the same bean flour (3.6% protein). Fig.14E is the chromatogram of defatted soy flour diluted to 8.7% protein, and Fig.14F is a chromatogram of a refined protein isolate from defatted soy flour (8.7% protein). Figure 14G is the chromatogram of yellow pea flour diluted to 7.4% protein, and Fig.14H is a chromatogram of a refined protein isolate from yellow pea flour (7.4% protein). Table 10 is a table of the HPLC peak areas plotted in Figure 13. The % of total peak area is reduced by nearly 100% in the samples tested.

[0395] Table 10.

[0396] Total peak area flour Refined protein isolate %reduce almond 2,864,059 26,005 99.1 kidney bean 5,028,531 52,010 99.0 Soybean 9,688,482 104,020 98.9 Yellow peas 19,355,660 208,040 98.9

[0397] GC chromatogram

[0398] We used gas chromatography (GC) analysis to quantify the volatile compounds present in flour and refined PI. Plants contain a wide variety of volatile compounds that may contribute to off-flavors and / or odors, at least to some extent. For five plant sources, protein isolate samples were compared to their original flour sources using gas chromatography. Samples were normalized to have the same amount of protein prior to analysis. The process in this example was able to reduce the total peak area from 88% to nearly 100% from flour to the final product.

[0399] Also tested in this example was the effectiveness of the process when purifying commercial protein isolates. Several commercial isolates (Puris, Roquette) were tested before and after the refining purification process. Figure 15 shows that the total peak area measured in the commercial PI (black bars) was greater than the area in the samples of the PI after the refining process (short dashed bars). The refining process purified protein resulted in the lowest peak area of ​​the sources tested. Table 11 below shows the GC peak areas in the original flour and refined PI samples. The % reduction ranged from 88% to nearly 100%. Fig.16A GC chromatogram showing defatted almond flour, and Fig. 16B Shown are GC chromatograms of refined PI from almond meal (both samples diluted to 10.7% protein). Fig. 16C shows the GC chromatogram of chickpea flour, and Fig.16D Shown are GC chromatograms of refined PI from chickpea flour (both samples diluted to 9.4% protein). Fig.16E GC chromatogram showing defatted soybean meal, and Fig.16F Shown are GC chromatograms of purified PI from defatted soy meal (both samples were diluted to 7.7% protein). Figure 16GShows the GC chromatogram of white kidney bean flour, and Fig.16H Shown are GC chromatograms of refined PI from white kidney bean flour (both samples were diluted to 3.4% protein). Fig.16I GC chromatogram showing yellow pea flour; Fig.16J GC chromatogram showing purified PI from yellow pea flour (on the same scale); and Figure 16K The GC chromatogram of purified PI from yellow pea flour is shown and compared with Fig.16J The GC chromatograms shown are identical but with an expanded view of the lower portion of the y-axis so that certain peaks are visible (both samples were diluted to 8.6% protein).

[0400] Table 11.

[0401] source Flour(total peak area) Refined PI (total peak area) % reduction in total area from flour to refined PI Defatted almonds 18,255,734 86,736 99.5 Chickpeas 68,212,786 2,336,181 96.6 Soybean 1,931,423 232,517 88.0 white kidney beans 8,228,243 377,364 95.4 Yellow peas 194,293,503 537,724 99.7

[0402] Chickpea flour and yellow pea flour had disproportionately large loading peaks. Most peaks did not appear in the PI. For each source tested, the total area reduction was greater than 88%. Refined protein isolates made from yellow pea and soy were compared to commercial protein isolates. Fig.17A -D shows the GC chromatogram of a commercial protein isolate sample compared with the same plant source after the refining process from flour to protein. Fig.17A The GC chromatogram of a refined protein isolate from yellow peas is shown and this can be compared to three commercial protein isolates (Nutralliance PPI-80 ( Fig. 17B ), Purple Pea 870( Fig. 17C ) and Roquette( Fig.17D Four samples were diluted to 8.6% protein (wt). Fig.17E -F shows a GC chromatogram of a refined protein isolate from soy flour compared to Now Sport soy protein isolate (both samples were diluted to 7.5% protein by weight).

[0403] Soy Isoflavones and Tannins

[0404] Isoflavones and tannins are two compounds known to activate bitter receptors. Therefore, this example evaluates the ability of the refined protein separation process to remove tannins and specific bitter isoflavones present in soy flour. The samples were normalized to protein for comparison, and it was clear that the presence of isoflavones detected in soy flour, daidzin, glycitein, and genistin was greatly reduced (95%, 87%, and to below the detection limit, respectively). Tannins, which produce astringent, bitter taste, were reduced by 84.7%. Other plant sources contain different amounts and types of isoflavones, so it is difficult to run this type of test on all eight sources. However, the results with soybeans indicate that the refined separation process is effective in reducing the levels of isoflavones and tannins in the final product. Table 12 shows the concentrations of known soy isoflavones in soy flour, water extracts, and refined protein isolates. The values ​​are normalized to protein % (result % (w / w) / protein % (w / w) x 100). If the values ​​are normalized to % dry mass, then daidzin is 0.014% dry mass, glycitein is below the detection limit, and genistin is 0.048% dry mass. For tannins, the value normalized to % dry mass is 0.18% dry mass.

[0405] Table 12.

[0406] test flour Extracts PI (Refined) %reduce Daidzin 0.377 0.731 0.018 95.2 Glycitein 0.048 0.088 ＜0.006 ND Genistin 0.473 0.744 0.061 87.1 Tannin, as tannic acid 1.507 3.139 0.231 84.7

[0407] Final protein characterization

[0408] Color (L*, a*, b* values)

[0409] One aspect of the refined protein separation process is the removal of color compounds. The production of pure white protein is desirable and can be achieved by using one or more embodiments of the refined protein separation process as disclosed herein.

[0410] The color of refined protein isolates obtained from commercial flour and other protein isolates on the market was measured using the CIELAB color space, where L* represents lightness (0 is black, and 100 is diffuse white), a* is a green / red scale (negative values ​​represent green, positive values ​​represent red), and b* is a blue / yellow scale (negative values ​​represent blue, positive values ​​represent yellow). The sample was prepared as follows: 3% protein (by weight), sunflower oil, sunflower lecithin, guar gum, gellan gum, and tap water. Prior to color analysis, the mixture was homogenized with a multitube mixer and allowed to settle.

[0411] The L* values ​​(related to the brightness of the sample) were higher in the refined protein isolates made from flour (ranging from 76 to 80) (see Table 13) than in the commercial protein isolates (ranging from 66 to 74) (see Table 14). The b values ​​(the higher the b value, the more yellow) were lower in the PIs made from flour.

[0412] Table 1 3.

[0413] PI from flour (refined) L* a* b* Chickpeas 76.9 -1.21 13.6 kidney bean 77.7 .1.13 5.91 white kidney beans 75.6 -1.13 4.51 broad bean 77.8 -1.70 1 1.7 defatted soybeans 77.8 .2.43 6.64 Defatted sesame seeds 78.6 -1.15 5.14 Defatted almonds 80.7 .0.90 5.50 Yellow peas 80.3 -0.67 9.86 scope 76 to 80 -2.5 to -0.7 4.5 to 12

[0414] Table 14.

[0415]

[0416] Emulsion stability

[0417] To compare changes in protein functionality, emulsion stability of proteins isolated from eight flour sources, as well as commercial whey and rice proteins, was measured (see Table 15). A lower instability index indicates a more stable emulsion. The instability index varied for different sources. Almond protein produced the least stable emulsion among those tested, while emulsions produced with soy protein and phaseol protein were more stable. Yellow pea, chickpea and broad bean had similar stabilities. This suggests that the refined protein isolation process can be used to produce proteins with different functionalities and can be used in end-product development.

[0418] In this example, the method used in measuring the emulsion stability of eight flour sources and whey and rice proteins is shown below. Measurements were made according to the following strategy:

[0419] 1.720 uL of 3.2% protein solution was mixed with 80 uL of sunflower oil and vortexed at speed 9 for 10 min.

[0420] 2. Allow to settle for 5 min. Take 200 uL of the emulsion and mix with 800 uL of SDS 0.1%.

[0421] 3. To measure the instability index in (by LUM GmbH), 350ul of sample was centrifuged at 1500rpm at 20°C for 3min.

[0422] Table 15.

[0423] Instability index (mean, n=6) sd PCV Yellow peas 0.443 0.02 4.7 Chickpeas 0.429 0.02 5.0 broad bean 0.476 0.02 4.5 white kidney beans 0.307 0.03 9.9 kidney bean 0.282 0.02 6.1 Soybean 0.244 0.03 10 Sesame 0.301 0.01 3.9 almond 0.511 0.03 6.4 Whey (n=3) 0.285 0.06 20 Rice (n=4) 0.466 0.05 11

[0424] Tasting Panel

[0425] To compare the mouthfeel profiles of refined PI with commercial protein isolates, emulsions were produced with each protein and submitted to a trained taste panel for analysis at Kansas State University. The source of the refined PI in this trial was a commercial PPI that was run through a refining process. Prior to use, the commercial protein was rinsed to achieve the same conductivity of the refined PI in order to remove the high levels of sodium present in most commercial pea protein isolates. Each solution consisted of protein (3.35% by weight), sunflower oil (1.48% by weight), and lecithin and gum to stabilize the emulsion. The emulsion ingredients are shown in Table 16.

[0426] Table 16.

[0427]

[0428] The taste panel evaluated the emulsions for 12 different flavor attributes, assigning the strongest flavor score of 5.0 to the flavor grains and processed. The commercial PPI had stronger "nutty," "woody," and "cereal" flavors (~1.0 point or higher), while the refined PI had a slightly higher score for "beany." The source of the refined PI was the commercial PI to which it was compared, so this trial demonstrated that the refined separation process removed multiple flavor attributes and produced a cleaner tasting product. Fig.18A The differences in taste attributes between emulsions produced with refined PI or rinsed commercial PPI (Roquette S85F) are shown. Fig.18B Individual graphs showing the taste attributes in emulsions produced with refined PI. Fig.18C Separate graphs showing the flavor attributes in emulsions produced with a rinsed commercial PPI (Roquette S85F) are shown. Table 16 shows the flavor attributes in the emulsions produced with a rinsed commercial PPI (Roquette S85F). Fig.18A The largest differences were in the flavor attributes of nutty, beany, woody, and cereal. Fig.19 (Aftertaste comparison between commercial protein and refined protein), Fig. 20 (Texture comparison between commercial and refined proteins) and Fig.21A and Fig. 21B Further evaluation is provided in (Appearance Comparison between Commercial and Refined Proteins).

[0429] Example 9: Production of purified protein on a commercial scale

[0430] 25 batches of refined protein were prepared for use as an ingredient in dairy milk analoques. Manufacturing was performed on a commercial scale as follows:

[0431] 1.600 kg of pea protein isolate (Roquette S85F) was slurried with water to a final solid loading of 12% (by wt%)

[0432] 2. Mix 3.2kg of NaOH into the pea protein and water slurry

[0433] 3. Then mix 46 kg of 34% (by weight) HCl into the pea protein and water slurry

[0434] 4. Then mix 17.8kg of anhydrous CaCl2 into the pea protein and water slurry

[0435] 5. The mixture was recirculated through 3 parallel decanter centrifuges, removing 22 gpm of liquid from the water slurry, and then adding 22 gpm of purified water for 2.5 hours until the bulk conductivity dropped to 2500 uS / cm.

[0436] 6. The slurry is then dewatered using a decanter centrifuge until the final dry solids are between 19-25%.

[0437] Pack the protein into a 275 gallon portable tank.

[0438] Final batch mass, L*a*b* color values, yield (in wt%), dry solids (wt%) and protein content (wt%) are shown below in Table 17. The color of the emulsion produced from the protein paste was determined as described in Example 8.

[0439] Table 17.

[0440]

[0441] Example 10: Chocolate spread made from refined protein

[0442] Dessert spreads were made with refined protein containing the following ingredients shown in Table 18 and the procedure is provided below.

[0443] Table 18.

[0444] Element supplier Weight(g) %(by weight) Organic High Heat Sunflower Oil Spectrum 145.6 29.1％ Refined protein N / A 165.9 33.2％ granulated sugar C&H 112.7 22.5％ Organic Agave Inulin Ciranda 49.1 9.8％ Rushmore Cocoa Powder Blommer 23.1 4.6％ Sunflower Seed Lecithin, Non-GMO Austrade 2.5 0.5％ Sea salt, refined Cargill 1.1 0.2％

[0445] program:

[0446] 1. Weigh out the ingredients.

[0447] 2. Add sugar to blender and blend on high speed for 30 seconds. Add dry solids to liquid and stir with spatula to combine.

[0448] 3. Add 5x weight (in ball bearings) to panning equipment and heat for 5 minutes.

[0449] 4. Add to the sugar coating equipment and run for 11 hours, scraping as needed.

[0450] 5. Store at 4°C

[0451] Example 11: Nutritional shake made with refined protein

[0452] The nutritional beverage was prepared with refined protein containing the following ingredients shown in Table 19 and the procedure was provided below.

[0453] program:

[0454] 1. Use the solution of Example 2 above to prepare plant-based dairy products.

[0455] 2. Mix the ingredients according to Table 20 below to form a powdered blend.

[0456] 3. Add powdered mix to plant-based milk and mix well.

[0457] Table 19.

[0458] Components supplier Code Weight(g) percentage Nutritional milkshake Plant-based dairy products, See the formulation in Table 4 of Example 2 354.88 88.53％ Powdered Mixture See the recipe in Table 20 46.00 11.47％ 400.88 100.00％

[0459] Table 20.

[0460]

[0461] Example 12. Other exemplary non-limiting embodiments

[0462] Further advantages of the claimed subject matter will become apparent from the following examples which describe certain embodiments of the claimed subject matter.

[0463] 1. A dairy analog comprising at least about 1% by weight protein obtained from one or more non-animal natural or modified non-animal natural sources and having a neutral color defined as an L* value of at least about 70, an a* value between about -1.5 and about +1.5, and a b* value between about -12 and about +12.

[0464] 2. The dairy analog of example 1, wherein the dairy analog further comprises lipids obtained from one or more non-animal natural or modified non-animal natural sources.

[0465] 3. The dairy analog of example 1 or 2, wherein at least one of said one or more non-animal natural or modified non-animal natural sources is a plant.

[0466] 4. The dairy product analog of example 3, wherein the plant is a leguminous plant.

[0467] 5. The dairy product analog of example 4, wherein the legume is pea.

[0468] 6. The dairy analog of example 1, wherein the dairy analog comprises less than about 1% by weight saturated fat.

[0469] 7. The dairy product analog of example 1, wherein the dairy product analog is substantially free of or free of one or more components selected from the group consisting of nut protein, cholesterol, and lactose.

[0470] 8. The dairy analog of example 1, wherein said dairy analog further comprises at least about 0.05% by weight calcium.

[0471] 9. A method for obtaining a certain yield of a neutral color refined protein component from a non-animal natural and / or modified non-animal natural source, comprising the steps of:

[0472] a) obtaining a protein preparation from said non-animal natural or modified non-animal natural source;

[0473] b) washing the protein preparation at a wash pH;

[0474] c) extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0475] d) separating the aqueous protein solution from non-aqueous components;

[0476] e) adding salt;

[0477] f) precipitating protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0478] g) separating the protein precipitate from non-precipitated components; and

[0479] h) washing the protein precipitate to obtain a substantial yield of a neutral-colored purified protein fraction.

[0480] 10. The method of example 9, wherein the non-animal natural or modified non-animal natural source is a plant.

[0481] 11. The method of example 10, wherein the plant is a leguminous plant.

[0482] 12. The method of example 11, wherein the leguminous plant is pea.

[0483] 13. The method of example 9, wherein the wash pH is between about 3 and about 5.

[0484] 14. The method of example 9, wherein the extraction pH is between about 8 and about 9.

[0485] 15. The method of example 9, wherein the added salt is calcium dichloride at a concentration between about 50 mM and about 100 mM.

[0486] 16. The method of example 9, wherein the precipitation pH is between about 5 and about 6.

[0487] 17. The method of example 9, wherein the method further comprises the step of adding between about 0.05% and about 12% by weight of a polysaccharide to the aqueous protein solution.

[0488] 18. The method of example 9, wherein the yield is between about 50% and about 90%.

[0489] 19. A neutral color refined protein component, wherein the neutral color refined protein component is obtained from a non-animal natural and / or modified non-animal natural source by one or more of the methods of Examples 9-18.

[0490] 20. The neutral colored refined protein component of Example 19, wherein the neutral colored refined protein component is a paste comprising between about 4% and about 25% (by weight) protein, between about 0.1% and about 1.5% (by weight) calcium, and between about 50% and about 90% (by weight) water.

[0491] 21. The neutral colored refined protein component of Example 19, wherein the neutral colored refined protein component is a dry powder comprising between about 70% and about 90% by weight protein and between about 2% and about 7% by weight calcium.

[0492] 22. The neutral colored refined protein component of Example 19, wherein the relative emulsifying activity of the neutral colored refined protein component is between about 0.1 and about 2 relative to the emulsifying activity of bovine serum albumin.

[0493] 23. The neutral colored refined protein fraction of example 19, wherein the neutral colored refined protein fraction comprises at least about 80% of visualized protein bands having a molecular weight of less than 200 kDa on a denaturing protein gel.

[0494] 24. A method for producing a dairy milk analog comprising the steps of:

[0495] a) obtaining at least one lipid component;

[0496] b) obtaining at least one neutral color refined protein component from a non-animal natural and / or modified non-animal natural source according to one or more of Examples 9-18;

[0497] c) mixing and blending an amount of the at least one lipid component, an amount of the at least one neutral-colored refined protein component, and an amount of a water component to produce a mixture; and

[0498] d) emulsifying the mixture to provide a dairy milk analog.

[0499] 25. The method of example 24, wherein the lipid component is obtained from a non-animal natural and / or modified non-animal natural source.

[0500] 26. The method of example 24 or 25, wherein at least one of the non-animal natural or modified non-animal natural sources is a plant.

[0501] 27. The method of example 26, wherein the plant is a leguminous plant.

[0502] 28. The method of example 27, wherein the leguminous plant is pea.

[0503] 29. The method of example 24 or 25, wherein the amounts of the lipid, protein, and water components are selected so as to provide a dairy milk analog comprising between about 1% and about 10% (by weight) protein obtained from a non-animal natural or modified non-animal natural source and between about 0% and about 4% (by weight) lipid obtained from a non-animal natural or modified non-animal natural source.

[0504] 30. The method of example 24, wherein the method further comprises the step of adding a carbohydrate component to the water component or mixture, wherein the carbohydrate component does not comprise lactose.

[0505] 31. The method of example 24, wherein the method further comprises the step of adding calcium to the water component or mixture.

[0506] 32. The method of example 24, wherein the method further comprises the step of sterilizing or pasteurizing the dairy milk analog.

[0507] 33. A method for producing a dairy yogurt analog, comprising the steps of:

[0508] a) obtaining at least one carbohydrate component;

[0509] b) obtaining at least one lipid component;

[0510] c) obtaining at least one neutral color refined protein component from a non-animal natural and / or modified non-animal natural source according to one or more of the methods of Examples 9-18;

[0511] d) blending the at least one carbohydrate component, the at least one lipid component, and the at least one protein component with a water component to produce a mixture;

[0512] e) heating the mixture;

[0513] f) emulsifying the mixture to produce an emulsified mixture;

[0514] g) cooling the emulsified mixture;

[0515] h) adding a fermentation microorganism to the emulsified mixture to produce a fermentation mixture; and

[0516] i) incubating the fermentation mixture at an elevated temperature until the fermentation mixture solidifies and acidifies to provide a dairy yogurt analog.

[0517] 34. The method of example 33, wherein the lipid component is obtained from a non-animal natural or modified non-animal natural source.

[0518] 35. The method of example 33 or 34, wherein at least one of the non-animal natural or modified non-animal natural sources is a plant.

[0519] 36. The method of example 35, wherein the plant is a leguminous plant.

[0520] 37. The method of example 36, wherein the leguminous plant is pea.

[0521] 38. The method of Example 33, wherein the amounts of the carbohydrate, lipid, protein and water components are selected so as to provide a dairy yogurt analog comprising between about 1% and about 10% (by weight) protein obtained from a non-animal natural or modified non-animal natural source and between about 0% and about 4% (by weight) lipid obtained from a non-animal natural or modified non-animal natural source.

[0522] 39. The method of example 33, wherein the carbohydrate component does not comprise lactose.

[0523] 40. The method of example 33, wherein the method further comprises the step of adding calcium to the water component or mixture.

[0524] 41. The method of example 33, wherein the fermenting microorganism is selected from the group consisting of Lactobacillus bulgaricus and Streptococcus thermophilus.

[0525] 42. The method of example 33, wherein the elevated temperature is 45°C.

[0526] 43. The method of example 33, wherein the method further comprises the step of sterilizing or pasteurizing the dairy yogurt analog.

[0527] 44. A method for obtaining a yield of a refined protein component from a non-animal native and / or modified non-animal native source, comprising the steps of:

[0528] a) obtaining a protein preparation from said non-animal natural or modified non-animal natural source;

[0529] b) washing the protein preparation at a wash pH;

[0530] c) extracting the protein preparation at an extraction pH to obtain an aqueous protein solution;

[0531] d) separating the aqueous protein solution from non-aqueous components;

[0532] e) adding salt;

[0533] f) precipitating protein from the aqueous protein solution at a precipitation pH to obtain a protein precipitate;

[0534] g) separating the protein precipitate from non-precipitated components; and

[0535] h) washing the protein precipitate to obtain a substantial yield of a refined protein component.

[0536] 45. The method of example 44, wherein the non-animal natural or modified non-animal natural source is a plant.

[0537] 46. ​​The method of example 44, wherein the plant is a leguminous plant.

[0538] 47. The method of example 44, wherein the leguminous plant is pea.

[0539] 48. The method of example 44, wherein the wash pH is between about 3 and about 5.

[0540] 49. The method of example 44, wherein the extraction pH is between about 8 and about 9.

[0541] 50. The method of example 44, wherein the added salt is calcium chloride at a concentration between about 50 mM and about 100 mM.

[0542] 51. The method of example 44, wherein the precipitation pH is between about 5 and about 6.

[0543] 52. The method of example 44, further comprising the step of adding between about 0.05% and about 12% by weight of a polysaccharide to the aqueous protein solution.

[0544] 53. The method of example 44, wherein the yield is between about 50% and about 90%.

[0545] 54. A refined protein component, wherein the refined protein component is obtained from a non-animal natural and / or modified non-animal natural source by the method of one or more of examples 44-53.

[0546] 55. The refined protein component of example 54, wherein the refined protein component is a paste comprising between about 4% and about 25% (by weight) protein, between about 0.1% and about 1.5% (by weight) calcium, and between about 50% and about 90% (by weight) water.

[0547] 56. The refined protein component of example 54, wherein the refined protein component is a dry powder comprising between about 70% and about 90% by weight protein and between about 2% and about 7% by weight calcium.

[0548] 57. The refined protein component of example 54, wherein the relative emulsifying activity of the refined protein component is between about 0.1 and about 2 relative to the emulsifying activity of bovine serum albumin.

[0549] 58. The refined protein fraction of example 54, wherein the refined protein fraction comprises at least about 80% of protein bands visualized on a denaturing protein gel having a molecular weight of less than 200 kDa.

[0550] 59. The refined protein component of one or more of examples 44-58, wherein the refined protein component is neutral in color.

[0551] 60. The refined protein component of one or more of examples 44-58, wherein the refined protein component is not neutral in color.

[0552] 61. A refined protein comprising between 5% and 97% by weight protein obtained from one or more non-animal natural or modified non-animal natural sources.

[0553] 62. The refined protein of example 61, wherein the weight percentage of the protein is between 20% and 90%.

[0554] 63. The refined protein of Example 61, wherein the weight percentage of the protein is between 30% and 85%.

[0555] 64. The refined protein of Example 61, wherein the weight percentage of the protein is between 40% and 80%.

[0556] 65. The refined protein of example 61, wherein the color is defined as an L* value between 60 and 90, an a* value between -6 and +6, and a b* value between -20 and +20.

[0557] 66. The refined protein of example 61, wherein the color is defined as an L* value between 65 and 85, an a* value between about -4 and about +4, and a b* value between -18 and +18.

[0558] 67. The refined protein of example 61, wherein the color is defined as an L* value of at least 65, an a* value of between at least -5 and +5, and a b* value of at least -16 to +16.

[0559] 68. The refined protein of example 61, wherein the color is defined as an L* value of at least 80, an a* value of at least -3 to +3, and a b* value of at least -14 to +14.

[0560] 69. The refined protein of one or more of examples 61-68, wherein the refined protein is a paste, a wet suspension, or a dry powder.

[0561] 70. The refined protein of one or more of examples 61-68, wherein the refined protein has a dry solids percentage of at least 5% by weight.

[0562] 71. The refined protein of one or more of examples 61-68, wherein the refined protein has a dry solids weight percentage of at least 10%.

[0563] 72. The refined protein of one or more of examples 61-68, wherein the refined protein has a dry solids weight percentage of at least 15%.

[0564] 73. The refined protein of one or more of examples 61-68, wherein the refined protein has a dry solids weight percentage of at least 20%.

[0565] 74. The refined protein of one or more of examples 61-68, wherein the refined protein has a dry solids weight percentage of at least 25%.

[0566] 75. The refined protein of one or more of examples 61-68, wherein the refined protein has a dry solids weight percentage of at least 30%.

[0567] 76. The refined protein of one or more of examples 61-75, wherein the refined protein further comprises calcium and the calcium to protein ratio is between 0.5% w / w and 5% w / w.

[0568] 77. The refined protein of one or more of examples 61-75, wherein the refined protein further comprises calcium and the calcium to protein ratio is between 1% w / w and 6% w / w.

[0569] 78. The refined protein of one or more of examples 61-75, wherein the refined protein further comprises calcium and the calcium to protein ratio is between 3% w / w and 8% w / w.

[0570] 79. The refined protein of one or more of examples 61-75, wherein the refined protein further comprises calcium and the calcium to protein ratio is between 5% w / w and 10% w / w.

[0571] 80. The refined protein of one or more of examples 61-79, wherein the refined protein is neutral in color.

[0572] 81. The refined protein of one or more of examples 61-79, wherein the refined protein is not neutral in color.

[0573] 82. The refined protein of one or more of examples 61-81, wherein the pH of the refined protein is between 4.5 and 11.

[0574] 83. The refined protein of one or more of examples 61-81, wherein the pH of the refined protein is between 6.5 and 10.

[0575] 84. The refined protein of one or more of examples 61-81, wherein the pH of the refined protein is between 5.5 and 8.

[0576] 85. The refined protein of one or more of examples 61-81, wherein the pH of the refined protein is between 5.7 and 6.7.

[0577] 86. The refined protein of one or more of examples 61-81, wherein the pH of the refined protein is at least 5.

[0578] 87. The refined protein of one or more of examples 61-81, wherein the pH of the refined protein is less than 9.

[0579] 88. The refined protein of one or more of examples 61-87, wherein the moisture content of the refined protein is between 3% and 90% by weight.

[0580] 89. The refined protein of one or more of examples 61-87, wherein the refined protein has a moisture content of at least 4% by weight.

[0581] 90. The refined protein of one or more of examples 61-87, wherein the refined protein has a moisture content of less than 80% by weight.

[0582] 91. The refined protein of one or more of examples 61-90, wherein the refined protein has a fat content of between 1% and 30% by weight.

[0583] 92. The refined protein of one or more of examples 61-90, wherein the refined protein has a fat content of at least 2% by weight.

[0584] 93. The refined protein of one or more of examples 61-90, wherein the refined protein has a fat content of less than 25% by weight.

[0585] 94. The refined protein of one or more of examples 61-93, wherein the carbohydrate content of the refined protein is between 0% and 50% by weight.

[0586] 95. The refined protein of one or more of examples 61-93, wherein the refined protein has a carbohydrate content of at least 0% by weight.

[0587] 96. The refined protein of one or more of Examples 61-93, wherein the carbohydrate content of the refined protein is less than 25% by weight.

[0588] 97. The refined protein of one or more of examples 61-96, wherein the starch content of the refined protein is between 0% and 10% by weight.

[0589] 98. The refined protein of one or more of Examples 61-96, wherein the starch content of the refined protein is at least 3% by weight.

[0590] 99. The refined protein of one or more of examples 61-96, wherein the refined protein has a starch content of less than 10% by weight.

[0591] 100. The refined protein of one or more of examples 61-99, wherein the phosphorus content of the refined protein is between 0% and 6% by weight.

[0592] 101. The refined protein of one or more of examples 61-99, wherein the refined protein has a phosphorus content of at least 0.1% by weight.

[0593] 102. The refined protein of one or more of examples 61-99, wherein the refined protein has a phosphorus content of less than 4% by weight.

[0594] 103. The refined protein of one or more of examples 61-102, wherein the refined protein has a sodium and / or potassium content of less than 0.5% by weight.

[0595] 104. The refined protein of one or more of examples 61-103, wherein the refined protein has an ash content between 0% and 20% by weight.

[0596] 105. The refined protein of one or more of examples 61-103, wherein the refined protein has an ash content of at least 1% by weight.

[0597] 106. The refined protein of one or more of examples 61-103, wherein the refined protein has an ash content of less than 10% by weight.

[0598] 107. The refined protein of one or more of examples 61-106, wherein the reducing power of the refined protein is between 5% and 50%.

[0599] 108. The refined protein of one or more of examples 61-106, wherein the reducing power of the refined protein is at least 6%.

[0600] 109. The refined protein of one or more of examples 61-106, wherein the reducing power of the refined protein is less than 46%.

[0601] 110. The refined protein of one or more of examples 61-109, wherein the total HPLC peak area of ​​the refined protein for total extractable soluble sugars and organic acids is between 20,000 and 250,000.

[0602] 111. The refined protein of one or more of examples 61-109, wherein the refined protein has a total extractable soluble sugars and organic acids of at least 22,000.

[0603] 112. The refined protein of one or more of examples 61-109, wherein the total extractable soluble sugars and organic acids of the refined protein is less than 240,000.

[0604] 113. The refined protein of one or more of examples 61-112, wherein the total peak area of ​​the refined protein is between 50,000 and 3,000,000 as measured by GC analysis of volatile compound components.

[0605] 114. The refined protein of one or more of examples 61-112, wherein the refined protein has a volatile compound fraction of less than 2,500,000.

[0606] 115. The refined protein of one or more of examples 61-114, wherein the isoflavone component of the refined protein is between 0% dry mass and 0.1% dry mass.

[0607] 116. The refined protein of one or more of examples 61-114, wherein the isoflavone component of the refined protein is less than 0.075% by dry mass.

[0608] 117. The refined protein of one or more of examples 61-116, wherein the tannin component of the refined protein is between 0% dry mass and 0.5% dry mass.

[0609] 118. The refined protein of one or more of examples 61-116, wherein the tannin component of the refined protein is less than 0.3% by dry mass.

[0610] 119. The refined protein of one or more of examples 61-118, wherein the refined protein has an instability index between 0.200 and 0.600.

[0611] 120. The refined protein of one or more of examples 61-118, wherein the refined protein has an instability index of at least 0.220.

[0612] 121. The refined protein of one or more of examples 61-118, wherein the refined protein has an instability index of less than 0.570.

[0613] 122. The refined protein of one or more of examples 61-121, wherein the refined protein has been produced in an amount of at least between 500 kg and 3000 kg, between 500 kg and 1000 kg, between 1000 kg and -2500 kg, and between 1000 kg and 3500 kg.

[0614] In addition, the present disclosure has been described with reference to specific embodiments. However, it will be readily apparent to those skilled in the art that it is feasible to implement the present disclosure in specific forms other than those described above. The above embodiments are merely illustrative and should not be considered restrictive. The scope of the present disclosure is given by the appended claims, rather than by the above description, and all variations and equivalents falling within the scope of the claims are intended to be included therein.

Claims

1. A refined protein component obtained from a vegetable source, comprising: between about 70% and about 90% protein by weight; and between about 0.1% and about 2.0% by weight calcium; wherein the refined protein component is derived from an alkaline extracted plant source; and The refined protein component is insoluble in an aqueous solution having a pH of less than about 6.

2. The refined protein component of claim 1 comprising between about 75% and about 90% protein by weight.

3. The refined protein component of claim 1, comprising at least 80% by weight protein.

4. The refined protein component of claim 1 comprising less than 10% by weight fat.

5. The refined protein component of claim 1, comprising less than 0.5% by weight potassium and sodium.

6. The refined protein component of claim 1 having a color defined as an L* (lightness) value of at least about 70, an a* (red / green) value between about -5 and about 5, and a b* (yellow / blue) value between about -16 and about 16.

7. The refined protein component of claim 1, wherein the refined protein component has an emulsifying activity of between about 0.1 and about 2 relative to the emulsifying activity of bovine serum albumin.

8. The refined protein fraction of claim 1, wherein at least about 80% of the protein bands visible to the naked eye have a molecular weight of less than 200 kDa when analyzed by denaturing polyacrylamide gel electrophoresis.

9. The refined protein component of claim 1, wherein the refined protein component is a dry powder.

10. The refined protein component of claim 1, wherein the wet protein paste of the refined protein component comprises: between about 4% and about 25% by weight protein; between about 0.1 and about 1.5 weight percent calcium; and Between about 50% and about 92% by weight water.

11. The refined protein component of claim 1, wherein the vegetable source comprises a leguminous plant.

12. The refined protein component of claim 1, wherein the vegetable source comprises pea.

13. The refined protein component of claim 1, wherein the vegetable source is selected from the group consisting of almonds, canola oil, oil palm, quinoa, rapeseed, sesame, and sunflower.

14. A refined protein powder obtained by extracting a vegetable protein preparation at an alkaline pH to obtain an aqueous protein solution, adding calcium to the aqueous protein solution, and precipitating at a pH of less than about 6, the refined protein powder comprising: between about 75% and about 90% protein by weight; between about 0.1% and about 2% by weight calcium; Less than 10% fat by weight; Less than 0.5% by weight of potassium; having a color defined as an L* (lightness) value of at least about 80, an a* (red / green) value between about -3 and about 3, and a b* (yellow / blue) value between about -14 and about 14; and The refined protein powder is insoluble in an aqueous solution having a pH of less than about 6.

15. The refined protein powder of claim 14, wherein the vegetable protein preparation is derived from a plant of the family Leguminosae.

16. The refined protein powder of claim 15, wherein the vegetable protein preparation is derived from peas.

17. The refined protein powder of claim 14, wherein the vegetable protein preparation is selected from the group consisting of almond, canola oil, oil palm, quinoa, rapeseed, sesame, and sunflower.

18. The refined protein powder of claim 14, wherein precipitating the protein at a pH of less than about 6 is performed in the presence of between about 10 mM and 50 mM calcium.

19. The refined protein powder of claim 14, wherein precipitating the protein at a pH of less than about 6 is performed in the presence of at least 0.05 grams of CaCl2 per gram of protein in the vegetable protein preparation.

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