Pongamia protein products and methods of production and use thereof

Abstract

Provided herein are protein-enriched pongamia compositions, including pongamia protein concentrates and isolates, which are suitable for consumption by animals, particularly humans. Also provided herein are various food and beverage products that can be fortified with such protein-enriched pongamia compositions. Further provided herein are methods of producing protein-enriched pongamia compositions, as well as methods of producing such protein-enriched pongamia compositions from pongamia beans or various forms of pongamia meal.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority and benefit to U.S. Provisional Applications 63 / 004,780, filed April 3, 2020; U.S. Provisional Applications 63 / 004,785, and U.S. Provisional Applications 63 / 004,792, filed April 3, 2020, the disclosures of which are incorporated herein by reference in their entirety. Technical Field

[0003] This disclosure generally relates to pongamia protein products. More specifically, this disclosure relates to pongamia compositions having a high protein content, such as pongamia protein concentrates or isolates, methods of producing them, and methods of using them in food and beverage products. Background Technology

[0004] With the global population projected to exceed 9 billion by 2050, the demand for edible protein continues to rise. The production of animal protein requires substantial resources, including land and water, and has a significant environmental impact. Plants offer an alternative and viable source of protein because they provide higher yields per acre and have a positive environmental impact. Consumer interest in sustainable, plant-based diets is expanding globally. In the United States, the plant-based food market was valued at approximately $4.5 billion in 2019. Nearly 55 million people in the U.S. are considered vegetarians or flexitarians and are seeking more plant-based protein-rich food options.

[0005] Millettia pinnata (L.) is a versatile tree that grows in the tropics and produces oil- and protein-rich legumes. Millettia has several advantageous characteristics: it is easy to grow, has a short generation time, and produces large quantities of oil- and protein-rich legumes. Millettia legumes, also known as Millettia oilseeds, contain approximately 35-40% oil and 20% protein. Millettia bean cake, a byproduct of oil extraction from Millettia legumes, provides a potential renewable source of plant protein for animal feed and human consumption. However, due to many inherent reactive chemical components such as thiamethoxam and thiamethoxam diketone, Millettia legumes and bean cake have a very bitter taste and are considered inedible. To produce edible Millettia protein products, these inherent reactive compounds must be removed or significantly reduced.

[0006] Currently, there is no commercially viable processing method for extracting and producing edible, clean-tasting concentrated aquaponica protein products from aquaponica beans. Therefore, a commercially viable method is needed to extract and remove intrinsic active chemical components, such as aquaponicain and aquaponicadione, from aquaponica beans and produce a value-added edible protein composition. Summary of the Invention

[0007] In some respects, this article provides water chestnut compositions with high protein content, such as water chestnut protein concentrates or isolates, which can be used as an edible protein source for animals, particularly humans. In some variations, the water chestnut protein products described herein possess functional properties, including, for example, solubility, viscosity, and emulsifying properties, which are comparable to or improved upon other commercial plant protein ingredients, such as soybean, pea, lupin, and sunflower. These water chestnut protein products can be used as useful ingredients in a variety of food and beverage products and address a significant unmet industry demand for emerging plant proteins with superior protein quality, excellent taste, and excellent texture.

[0008] In some aspects, a protein-rich *Phellodendron amurense* composition is provided, comprising at least 50% *Phellodendron amurense* protein based on dry weight. In some aspects, a protein-rich *Phellodendron amurense* composition is provided, comprising at least 70% *Phellodendron amurense* protein based on dry weight. In some embodiments, the composition is a *Phellodendron amurense* protein concentrate. In other embodiments, the composition is a *Phellodendron amurense* protein isolate.

[0009] In some aspects, a method for producing a protein-rich water pomegranate composition is provided, comprising: preparing an aqueous slurry of water pomegranate meal; adjusting the pH of the aqueous slurry to a pH between 8 and 10; separating the slurry into a protein liquid fraction and an insoluble wet filter cake fraction; neutralizing, concentrating, and / or pasteurizing the protein liquid fraction; and drying the protein liquid fraction to provide a protein-rich water pomegranate composition.

[0010] In some aspects, a method for producing a protein-rich water citrus composition is provided, comprising: preparing an aqueous slurry of water citrus pomace; adjusting the pH of the aqueous slurry to a pH between 8 and 10; separating the slurry into a protein liquid fraction and an insoluble wet filter cake fraction; precipitating a portion of the water citrus protein from the liquid fraction by adjusting the pH to a pH between 3.5 and 4.5 to obtain purified water citrus protein; washing, neutralizing, and / or pasteurizing the purified water citrus protein; and drying the purified water citrus protein to provide a protein-rich water citrus composition.

[0011] In some aspects, a method for producing a protein-rich water citrus composition is provided, comprising: preparing an aqueous slurry of water citrus pulp; adjusting the pH of the aqueous slurry to a pH between 6 and 10; separating the slurry into a protein liquid fraction and an insoluble wet filter cake fraction; passing the protein liquid fraction through a membrane system to obtain a retentate comprising water citrus protein; washing, neutralizing, and / or pasteurizing the retentate; and drying the retentate to provide a protein-rich water citrus composition.

[0012] In one aspect, a protein-rich water-rich wampee composition is provided, produced according to the method described herein.

[0013] In some aspects, various products incorporating any of the protein-rich water lily compositions described herein are also provided. In some embodiments, the product is a food product, a beverage product, or a dietary supplement product. In some variations, the product is a baked good, a protein supplement, a protein bar, or a non-dairy beverage. In still other variations, the product is a medicinal food, infant formula, a cosmetic, or a pharmaceutical product.

[0014] In one aspect, a protein-rich aquamarine component is provided, comprising at least 40% aquamarine protein based on dry weight, wherein the component has: (i) less than 500 ppm of aquamarine glycoside; (ii) less than 500 ppm of aquamarine dione; (iii) less than 500 ppm of a combination of aquamarine glycoside and aquamarine dione; and wherein the component has less than or equal to 40% carbohydrates based on dry weight. In some embodiments of this aspect, the protein-rich aquamarine component comprises at least 70% aquamarine protein based on dry weight, wherein the component has: (i) less than 500 ppm of aquamarine glycoside; (ii) less than 500 ppm of aquamarine dione; (iii) less than 500 ppm of a combination of aquamarine glycoside and aquamarine dione; and wherein the component has less than or equal to 15% carbohydrates based on dry weight.

[0015] In some embodiments, the component has (i) in 100s -1 (ii) the shear rate, viscosity between approximately 2 mPa*s and approximately 100 mPa*s; (iii) the foaming capacity of 0.1% protein solution volume between approximately 100% and approximately 200%; and (iv) at least approximately 0.2 g / cm³. 3(iv) Bulk density; at pH 7, at least about 35% protein solubility; (v) Median emulsion droplet size less than or equal to about 5 μm; (vi) Median emulsion droplet size less than or equal to about 5 μm after 7 days of storage; (vi) Water-holding capacity of at least about 1.5 g of water per gram of protein-rich water-rich yellow peel component; (vii) Oil-holding capacity of at least about 1.5 g of oil per gram of protein-rich water-rich yellow peel component; (viii) Minimum gelling concentration of at least about 10 g of protein-rich water-rich yellow peel component per 100 g; (ix) Powder dispersibility of at least about 10%; (x) Neutral and without bitterness; or any combination thereof (i)-(x).

[0016] In other embodiments, the component has: (i) in 100s -1 (ii) the shear rate, viscosity between approximately 2 mPa*s and approximately 100 mPa*s; (iii) the foaming capacity of a 0.1% w / v water solution of citrus aurantiacus protein at approximately 100% to approximately 200% of its volume; and (iii) a foaming capacity of at least approximately 0.2 g / cm³. 3 (iv) Bulk density; at pH 7, at least about 35% protein solubility; (v) Median emulsion droplet size less than or equal to about 5 μm; (vi) Median emulsion droplet size less than or equal to about 5 μm after 7 days of storage; (vii) Neutral and non-bitter; or any combination thereof (i)-(vii).

[0017] In other embodiments, the component has: (i) in 100s -1 (ii) the shear rate, viscosity between approximately 2 mPa*s and approximately 100 mPa*s; (iii) the foaming capacity of a 0.1% w / v protein solution volume between approximately 100% and approximately 200%; and (iii) at least approximately 0.2 g / cm³. 3 (iv) Bulk density; at pH 7, at least about 35% protein solubility; (v) Median emulsion droplet size less than or equal to about 5 μm; (vi) Median emulsion droplet size less than or equal to about 5 μm after 7 days of storage; (vii) Water-holding capacity of at least about 1.5 g of water per gram of protein-rich water-rich yellow peel component; (viii) Minimum gelling concentration of at least about 10 g of protein-rich water-rich yellow peel component per 100 g; (ix) Neutral and without bitterness; or any combination of (i)-(ix).

[0018] In other embodiments, the component has (i) at least about 0.2 g / cm³. 3(i) bulk density; (ii) protein solubility of at least about 35% at pH 7; (iii) water-holding capacity of at least about 1.5 g of water per gram of protein-rich water-rich yellow peel component; (iv) oil-holding capacity of at least about 1.5 g of oil per gram of protein-rich water-rich yellow peel component; (v) minimum gelling concentration of at least about 10 g of protein-rich water-rich yellow peel component per 100 g; (vi) neutral and without bitterness; or any combination thereof (i)-(vi).

[0019] In some embodiments, the ingredient has: (i) a foaming capacity between about 100% and about 200% of the volume of a 0.1% w / v protein solution; (ii) a minimum gelling concentration of at least about 7g of protein-rich water phellodendron ingredient per 100g; (iii) neutral and without bitterness; or any combination thereof (i)-(iii).

[0020] In one aspect, this article provides a method for producing a protein-rich water citrus composition, comprising: preparing an aqueous slurry of water citrus pulp, wherein the water citrus pulp is defatted and debittered and has (i) less than 500 ppm of water citrusin; or (ii) less than 500 ppm of water citrus diketone; or (iii) less than 500 ppm of a combination of water citrusin and water citrus diketone; adjusting the pH of the aqueous slurry to a pH between 6 and 10; separating the slurry into a protein liquid fraction and an insoluble wet filter cake fraction; neutralizing, concentrating, and / or pasteurizing the protein liquid fraction; and drying the protein liquid fraction to provide a protein-rich water citrus composition.

[0021] In another aspect, this article provides a method for producing a protein-rich water hibiscus component, comprising: preparing an aqueous slurry of water hibiscus meal, wherein the water hibiscus meal is defatted and debittered and has (i) less than 500 ppm of water hibiscus glycoside; or (ii) less than 500 ppm of water hibiscus dione; or (iii) less than 500 ppm of a combination of water hibiscus glycoside and water hibiscus dione; adjusting the pH of the aqueous slurry to a pH between 6 and 10; separating the slurry into a protein liquid fraction and an insoluble wet filter cake fraction; precipitating at least a portion of the water hibiscus protein from the protein liquid fraction to obtain a purified water hibiscus protein solid; neutralizing and pasteurizing the purified water hibiscus protein solid; and drying the purified water hibiscus protein solid to provide a protein-rich water hibiscus component.

[0022] In another aspect, this article provides a method for producing a protein-rich water pomegranate component, comprising: preparing an aqueous slurry of water pomegranate meal; adjusting the pH of the aqueous slurry to a pH between 6 and 10; separating the slurry into a protein liquid fraction and an insoluble wet filter cake fraction; passing the protein liquid fraction through a membrane system to obtain a retainer comprising water pomegranate protein; optionally washing, neutralizing, and / or pasteurizing the retainer; and drying the retainer to provide a protein-rich water pomegranate component. Attached Figure Description

[0023] This application can be understood by referring to the following description in conjunction with the accompanying drawings.

[0024] Figure 1 An overview of an exemplary method for producing a water-yellow peel composition with a high protein content from water-yellow peel beans is provided.

[0025] Figure 2A An exemplary method for producing a water yellow peel protein concentrate from defatted, debittered water yellow peel meal by solubilization is described.

[0026] Figure 2B An exemplary method for producing a water yellow peel protein isolate from defatted, debittered water yellow peel meal by isoelectric precipitation is described.

[0027] Figure 2C An exemplary method for producing a water yellow peel protein isolate from defatted, debittered water yellow peel meal via membrane filtration is described.

[0028] Figure 3 The solubility curves of water yellow peel protein in freeze-dried water yellow peel protein concentrate at different pH values ​​were plotted.

[0029] Figures 4A-4C The description shows the solubility of an exemplary aqueous yellow peel protein concentrate. Figure 4A ), 100s -1 viscosity at shear rate ( Figure 4B ) Targeting commercial proteins (soy, pea, and lupin) and regarding emulsions ( Figure 4C A chart comparing the functional properties of commercial proteins (soybean, pea, and sunflower).

[0030] Figure 4D-4F The solubility of an exemplary aqueous yellow peel protein isolate is depicted. Figure 4D ), 100s -1 viscosity at shear rate ( Figure 4E ) Targeting commercial proteins (soy, pea, and lupin) and regarding emulsions ( Figure 4F A chart comparing the functional properties of commercial proteins (soybean, pea, and sunflower).

[0031] Figures 5A-5D The protein composition resolved by SDS-PAGE was depicted, showing the molecular weight distribution of various water yellow-skinned bean proteins to illustrate the stability and integrity of the method and to distinguish water yellow-skinned bean proteins relative to soybean proteins.

[0032] Figure 6A and 6B The viscosity and emulsification properties of the water yellow peel protein isolate produced on a pilot scale were described compared with those of pea and soybean protein isolates. Detailed Implementation

[0033] The following description sets forth exemplary methods, parameters, etc. However, it should be understood that this description is not intended to be a limitation on the scope of this disclosure, but is provided as a description of exemplary embodiments.

[0034] In some aspects, this document provides compositions of *Phellodendron amurense* with high protein content, comprising, for example, *Phellodendron amurense* protein concentrates or isolates, said compositions being usable as an alternative plant-based protein source for consumption by animals, including, in particular, humans. In some aspects, this document provides methods for producing such protein-rich *Phellodendron amurense* compositions, and methods for using such compositions in various food and beverage products.

[0035] In some respects, the protein-rich water lily composition provided herein is a protein-rich water lily component. As used herein, "protein-rich water lily composition" may be interchangeably referred to as "protein-rich water lily component." The protein-rich water lily component of this disclosure differs from water lily bean cake, water lily meal, or water lily powder in that the water lily component provided herein is rich in protein, while other components such as carbohydrates and fats are reduced in amount relative to those naturally present in water lily beans, bean cake, meal, or powder. Water lily bean cake, meal, and powder are typically prepared by detoxifying (or debittering) and defatting the source water lily beans, but without substantially increasing their protein content or reducing their carbohydrate content. For example, typical water lily beans contain approximately 25% protein based on dry weight. Typical methods of detoxifying and / or defatting water lily beans produce water lily bean cake, meal, or powder with a protein content of 30-35%.

[0036] In addition, the protein-rich components of water yellow peel exhibit many physical properties that reflect the enrichment of protein and the reduction of fat and carbohydrates, which makes them suitable for a wider range of food applications than water yellow peel bean cakes, meals, or powders.

[0037] A protein-rich water citrus fruit composition

[0038] The composition and properties of the protein-rich water hyacinth peel composition are described in further detail below. Surprisingly, the functional properties of the protein-rich water hyacinth peel composition described herein, such as solubility, viscosity, and emulsifying properties, are comparable, if not superior, to those of commercial plant protein ingredients such as soybean, pea, lupin, and sunflower. These properties suggest that this product could be found in a variety of food and beverage products.

[0039] Water yellow peel protein content

[0040] The protein-rich water citrus composition provided herein has a high water citrus protein content, which includes the water citrus protein content relative to the water citrus meal from which the rich composition is obtained.

[0041] In some variations, the protein-rich *Phellodendron amurense* composition has *Phellodendron amurense* protein content based on a dry weight of at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 95%. In some variations, the protein-rich *Phellodendron amurense* composition has *Phellodendron amurense* protein content based on a dry weight of between 50% and 99%, between 50% and 95%, between 50% and 90%, between 50% and 85%, between 50% and 80%, between 50% and 75%, between 45% and 70%, between 45% and 60%, between 40% and 70%, between 40% and 95%, or between 45% and 90%.

[0042] In certain variants, the protein-rich water hibiscus composition is a water hibiscus protein concentrate having at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, or at least 65% based on dry weight; or between 45% and 70%, between 45% and 60%, between 50% and 70%, between 55% and 70%, between 55% and 65%, between 55% and 60%, between 60% and 70%, or between 65% and 70% water hibiscus protein.

[0043] In other variations, the protein-rich *Phellodendron amurense* composition is a *Phellodendron amurense* protein isolate having, based on dry weight, at least 70%, at least 75%, at least 80%, at least 85%, or at least 95%; or between 70% and 95%, between 75% and 95%, between 80% and 95%, between 85% and 95%, between 90% and 95%, between 70% and 90%, between 75% and 90%, between 80% and 90%, between 85% and 90%, between 70% and 85%, between 75% and 85%, between 80% and 85%, between 70% and 80%, or between 75% and 80% of *Phellodendron amurense* protein.

[0044] In some of the aforementioned variations, the protein-rich water jasmine composition has at least 1.1 times, at least 1.25 times, at least 1.5 times, at least 2 times, at least 2.5 times, at least 3 times, at least 3.5 times, or at least 4 times more water jasmine protein than the water jasmine meal from which the rich composition is obtained. In some variations, the protein-rich water jasmine composition has between 1.25 and 5 times more water jasmine protein than the water jasmine meal from which the rich composition is obtained.

[0045] Protein solubility

[0046] Protein solubility can be measured using any suitable technique known in the art. For example, in one variation, solubility is measured according to the protocol described in Example 4 below.

[0047] In some embodiments, at least 35%, 40%, 50%, 60%, 70%, 75%, 80%, or 85% of the protein in the protein-rich *Phellodendron amurense* composition is soluble in water with a pH greater than or equal to pH 6. In some variations, at least 35%, 40%, 50%, 60%, 70%, 75%, 80%, or 85% of the protein in the protein-rich *Phellodendron amurense* composition is soluble in water with a pH of 7. In other embodiments, less than or equal to 50%, less than or equal to 45%, less than or equal to 40%, or less than or equal to 30% of the protein in the protein-rich *Phellodendron amurense* composition is soluble in water with a pH between 3 and 5. In some variations, less than or equal to 50%, less than or equal to 45%, less than or equal to 40%, or less than or equal to 30% of the protein in the protein-rich *Phellodendron amurense* composition is soluble in water with a pH of 4.5.

[0048] Carbohydrate content

[0049] In conjunction with the enrichment of proteins in the protein-rich water pomegranate composition provided herein, the protein-rich water pomegranate composition has a reduced carbohydrate content, which includes the carbohydrate content relative to the water pomegranate meal from which the enriched composition is obtained.

[0050] In some embodiments, the protein-rich water citrus composition has carbohydrates of less than or equal to about 50%, less than or equal to about 40%, less than or equal to about 35%, less than or equal to about 30%, less than or equal to about 25%, less than or equal to about 20%, less than or equal to about 15%, or less than or equal to about 10% by dry weight.

[0051] In some embodiments, the protein-rich *Phellodendron amurense* composition has at least 40% *Phellodendron amurense* protein based on dry weight and less than or equal to about 50% carbohydrates based on dry weight. In some embodiments, the protein-rich *Phellodendron amurense* composition has at least 40% *Phellodendron amurense* protein based on dry weight and less than or equal to about 40% carbohydrates based on dry weight. In other embodiments, the protein-rich *Phellodendron amurense* composition has at least 70% *Phellodendron amurense* protein based on dry weight and less than or equal to about 20% carbohydrates based on dry weight. In some other embodiments, the protein-rich *Phellodendron amurense* composition has at least 70% *Phellodendron amurense* protein based on dry weight and less than or equal to about 15% carbohydrates based on dry weight.

[0052] Fat content

[0053] In some embodiments, the protein-rich water lily composition has a fat content of less than or equal to 0.5%, less than or equal to 0.75%, or less than or equal to 1% based on dry weight. In other embodiments, the protein-rich water lily composition has a fat content of less than or equal to 4%, less than or equal to 3%, less than or equal to 2%, or less than or equal to 1% based on dry weight. In some embodiments, the protein-rich water lily composition has a fat content between 0.5% and 4%, between 0.5% and 3%, between 0.5% and 2%, between 0.5% and 1%, between 0.75% and 4%, between 0.75% and 3%, between 0.75% and 2%, between 0.75% and 1%, between 1% and 4%, between 1% and 3%, between 1% and 2%, or between 0% and 1% based on dry weight.

[0054] In other embodiments of the aforementioned protein-rich water citrus peel composition, depending on the composition of the water citrus peel meal used, the protein-rich water citrus peel composition may have:

[0055] (i) Based on fat content of less than 4%, less than 3%, less than 2%, or less than 1% by dry weight; or

[0056] (ii) Bitter compounds less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm or less than 1 ppm.

[0057] Or both (i) and (ii).

[0058] Furan flavonoids

[0059] Bitter compounds refer to compounds naturally found in the citrus aurantium that have a bitter taste. In some embodiments, the bitter taste can be attributed to furan flavonoids such as citrus quercetin and citrus quercetin dione.

[0060] In some of the aforementioned variations, the bitter compounds present in the protein-rich water croton composition may include water crotonin and / or water croton dione. Therefore, in some of the foregoing embodiments, the protein-rich water flavour composition comprises: (i) less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm of water flavourin; (ii) less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm of water flavoured dione; or (iii) a combination of water flavourin and water flavoured dione less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm, or less than 1 ppm.

[0061] In some variations, the content of chlorophyllin and / or chlorophyllide dione in the protein-rich *Phellodendron amurense* compositions provided herein is determined by liquid chromatography. Analytical methods for determining the content of chlorophyllin and / or chlorophyllide dione may include solvent extraction of the *Phellodendron amurense* sample followed by liquid chromatography analysis. In some variations, the extraction solvent includes alkyl esters of alkanonic acids. In one variation, the extraction solvent includes ethyl acetate. In some embodiments, the liquid chromatography analysis may include high-performance liquid chromatography and mass spectrometry (e.g., MS or MS / MS) or ultraviolet detection (e.g., UV, UV / Vis, or DAD).

[0062] In some variations, solvent extraction may involve microwave-assisted solvent extraction. For example, an exemplary analytical method may include microwave-assisted extraction of hydroflavin and hydroflavin dione using ethyl acetate to collect an extract for subsequent high-performance liquid chromatography and mass spectrometry (HPLC-MS) or UV spectrophotometry. A hydroflavin sample is added to a microwave extraction tube. The extraction solvent is then added to the sample tube and vortexed to mix. Next, the sample is extracted using a microwave extractor. Once cooled, the supernatant is vacuum filtered to remove particulates. Alternatively, the supernatant may be centrifuged to remove particulates. The extracted sample can then be analyzed by HPLC (and detected by mass spectrometry or UV spectrophotometry) to determine the levels of hydroflavin and hydroflavin dione present in the sample.

[0063] In some embodiments, the protein-rich water hibiscus composition comprises at least 40% water hibiscus protein based on dry weight, wherein the components have: (i) less than 500 ppm of water hibiscus linolenic acid; (ii) less than 500 ppm of water hibiscus dione; (iii) less than 500 ppm of a combination of water hibiscus linolenic acid and water hibiscus dione; and wherein the components have less than or equal to 40% carbohydrates based on dry weight.

[0064] In some other embodiments, the protein-rich water hibiscus composition comprises at least 70% water hibiscus protein based on dry weight, wherein the components have: (i) less than 500 ppm of water hibiscus linolenic acid; (ii) less than 500 ppm of water hibiscus dione; (iii) less than 500 ppm of a combination of water hibiscus linolenic acid and water hibiscus dione; and wherein the components have less than or equal to 15% carbohydrates based on dry weight.

[0065] Relative amino acid spectrum

[0066] In some of the foregoing embodiments, the water-wheat peel protein present in the protein-rich water-wheat peel composition has a protein-based relative amino acid profile, which includes:

[0067] (i) at least about 0.5% methionine,

[0068] (ii) At least about 1% tryptophan,

[0069] (iii) At least about 1% cysteine,

[0070] (iv) At least about 2% histidine,

[0071] (v) At least about 3% threonine,

[0072] (vi) At least about 3% isoleucine,

[0073] (vii) At least about 3% tyrosine,

[0074] (viii) At least about 3% alanine,

[0075] (ix) At least about 3% glycine,

[0076] (x) at least about 4% valine,

[0077] (xi) at least about 5% proline,

[0078] (xii) at least about 5% serine,

[0079] (xiii) At least about 5% arginine,

[0080] (xiv) at least about 6% phenylalanine,

[0081] (xv) at least about 8% lysine,

[0082] (xvi) at least approximately 9% leucine,

[0083] (xvii) at least about 12% aspartic acid,

[0084] (xviii) At least about 15% glutamic acid,

[0085] Or any combination of (i)-(xviii).

[0086] In some embodiments, the protein-rich water jasmine composition has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, at least 6% phenylalanine, or any combination thereof. In some variations, the protein-rich water jasmine composition has a relative amino acid profile comprising at least 15% glutamic acid. In other variations, the protein-rich water jasmine composition has a relative amino acid profile comprising at least 12% aspartic acid. In still other variations, the protein-rich water jasmine composition has a relative amino acid profile comprising at least 9% leucine. In still other variations, the protein-rich water jasmine composition has a relative amino acid profile comprising at least 8% lysine. In still still other variations, the protein-rich water jasmine composition has a relative amino acid profile comprising at least 6% phenylalanine. In certain variants, the protein-rich water citrus composition has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, and at least 6% phenylalanine.

[0087] Amino acid score

[0088] In a further embodiment, the protein-rich water chestnut composition is characterized by an amino acid score, which compares the amount of amino acids present in the composition to the amount of the same amino acids present in a reference composition. The amino acid (AA) score is calculated as follows:

[0089] Amino acid (AA) score = (mg of limited essential amino acids in 1g of test protein) / (mg of the same essential amino acids in 1g of reference protein) × 100.

[0090] In some embodiments, the protein-rich water lily composition has an AA score of at least one of the following: greater than or equal to 70, greater than or equal to 80, greater than or equal to 90, greater than or equal to 100, greater than or equal to 125, or greater than or equal to 150: a combination of cysteine ​​and methionine, histidine, isoleucine, leucine, lysine, threonine, tryptophan, a combination of tyrosine and phenylalanine, or valine. In some embodiments, the protein-rich water lily composition has an AA score of various of the following: greater than or equal to 70, greater than or equal to 80, greater than or equal to 90, greater than or equal to 100, greater than or equal to 125, or greater than or equal to 150: a combination of cysteine ​​and methionine, histidine, isoleucine, leucine, lysine, threonine, tryptophan, a combination of tyrosine and phenylalanine, and valine.

[0091] The protein digestibility-corrected amino acids score (PDCAAS) is a known method in the art for assessing protein quality based on human amino acid requirements and their ability to digest proteins. A value of "1" is the highest, and "0" is the lowest. The formula for calculating PDCAAS, as specified by the FAO / WHO / UNU Expert Consultation, is as follows:

[0092] PDCAAS = Minimum Essential Amino Acid Ratio x Actual Fecal Digestibility (%)

[0093] The minimum essential amino acid ratio is derived as follows:

[0094] (mg of the limited essential amino acids in 1g of test protein) / (mg of the same essential amino acids in 1g of reference protein)

[0095] In some embodiments, the protein-rich *Phellodendron amurense* composition described herein has a relatively high PDCAAS value. In some variations, the protein-rich *Phellodendron amurense* composition has a PDCAAS of at least 0.7, at least 0.75, at least 0.8, at least 0.85, at least 0.9, or at least 0.95. In some variations, the protein-rich *Phellodendron amurense* composition has a PDCAAS between 0.7 and 0.95, between 0.75 and 0.95, between 0.8 and 0.95, between 0.85 and 0.95, between 0.9 and 0.95, between 0.75 and 0.9, between 0.8 and 0.9, between 0.85 and 0.9, or between 0.8 and 0.85.

[0096] molecular weight

[0097] The molecular weight distribution of proteins present in the protein-rich water hibiscus composition can be determined using any suitable technique known in the art. For example, in one variant, the molecular weight is determined according to the scheme described in Example 3 below.

[0098] The protein-rich *Phellodendron amurense* compositions disclosed herein are obtained from *Phellodendron amurense* meal derived from *Phellodendron amurense* beans. Because they are derived from *Phellodendron amurense* oilseeds, in contrast to compositions obtained from, for example, *Phellodendron amurense* leaves, the protein-rich *Phellodendron amurense* compositions provided herein contain seed-stored proteins, and are characterized by the presence of these seed-stored proteins. In some embodiments, the protein-rich *Phellodendron amurense* compositions of this disclosure can be distinguished from protein compositions derived from other plant sources, such as peas or soybeans, by the molecular weight distribution of the proteins present within the protein-rich *Phellodendron amurense* compositions.

[0099] In some embodiments, the protein-rich *Phellodendron amurense* compositions described herein comprise proteins with different molecular weights. In some variations, the *Phellodendron amurense* protein concentrate or isolate has an average molecular weight greater than or equal to 10,000, 15,000, 20,000, 25,000, 30,000, 35,000, 40,000, 45,000, 50,000, or 55,000 Daltons. In other variations, the protein-rich *Phellodendron amurense* compositions have an average molecular weight less than or equal to 250,000, 200,000, 175,000, 150,000, 130,000, 120,000, 110,000, 100,000, 90,000, 80,000, 70,000, 60,000, or 55,000 Daltons. In certain variations, the protein-rich water citrus composition has an average molecular weight falling within a range, where any of the aforementioned weights can be used as the upper or lower limit of the range. For example, in one variation, the protein-rich water citrus composition has an average molecular weight between 55,000 Daltons and 72,000 Daltons. In other variations, the protein-rich water citrus composition has an average molecular weight between 5,000 Daltons and 250,000 Daltons.

[0100] In some embodiments, the protein-rich *Phellodendron amurense* composition includes seed storage proteins. In some embodiments, the protein-rich *Phellodendron amurense* composition includes seed storage proteins, wherein about 30-40% of the protein present is a protein having a molecular weight between about 45 kDa and about 70 kDa as determined by SDS-PAGE. In other embodiments, the protein-rich *Phellodendron amurense* composition includes significant seed storage proteins having a molecular weight of about 170-250 kDa, about 115-160 kDa, about 45-70 kDa, about 19-25 kDa, about 14-17 kDa, or about 10-13 kDa, or any combination thereof.

[0101] In some variations, the molecular weights of the protein-rich water pomegranate compositions, components, concentrates, and isolates provided herein were determined by SDS-PAGE according to the scheme described in Example 3.

[0102] Viscosity

[0103] The protein-rich water-wampee compositions presented in this article can also be characterized by their viscosity when prepared in solution. The viscosity of such solutions rich in water-wampee protein is suitable for certain food product applications, such as beverages, where viscosity can affect the overall thickness or thinness of the food product. Higher viscosity is more suitable for certain food products, such as yogurt, while lower viscosity is more suitable for high-protein beverages.

[0104] Viscosity can be measured using any suitable technique known in the art. For example, in one variation, the viscosity of the protein solution is measured using a rheometer according to the procedure described in Example 4 below.

[0105] In some embodiments, the protein-rich water chestnut composition has different viscosities. In some variations, the protein-rich water chestnut composition has a viscosity of 100s. -1 The viscosity was measured at a shear rate of at least 2 mPa·s, at least 3 mPa·s, or at least 4 mPa·s. In some variants, the protein-rich water-based yellow peel composition exhibited a viscosity of 100 s⁻¹. -1 The viscosity was measured at a shear rate less than or equal to 100 mPa*s, less than or equal to 75 mPa*s, less than or equal to 50 mPa*s, or less than or equal to 25 mPa*s. In other variations, the protein-rich water-based yellow peel composition exhibited a viscosity at 100 mPa*s. -1 The viscosity measured at shear rates less than or equal to about 10 mPa*s, less than or equal to 8 mPa*s, less than or equal to 7 mPa*s, less than or equal to 6 mPa*s, or less than or equal to 5 mPa*s. In other variations, the protein-rich water-based yellow peel composition exhibits a viscosity of less than 100 s⁻¹. -1 Viscosities at shear rates between approximately 2 mPa*s and approximately 100 mPa*s, between approximately 2 mPa*s and approximately 75 mPa*s, between approximately 2 mPa*s and approximately 50 mPa*s, between approximately 2 mPa*s and approximately 25 mPa*s, between approximately 2 mPa*s and approximately 10 mPa*s, between approximately 5 mPa*s and approximately 10 mPa*s, or between approximately 7 mPa*s and approximately 10 mPa*s.

[0106] Among other variations, the protein-rich water-wampee composition has a 50s -1 The viscosity was measured at a shear rate of at least 2 mPa*s, at least 4 mPa*s, at least 6 mPa*s, or at least 8 mPa*s. In some variants, the protein-rich water-based yellow peel composition had a viscosity of at least 50 mPa*s. -1 The viscosity was measured at a shear rate less than or equal to 15 mPa*s, less than or equal to 12 mPa*s, or less than or equal to 10 mPa*s. In other variations, the protein-rich water-based yellow peel composition exhibited a viscosity of 10 mPa*s. -1 The viscosity was measured at a shear rate of at least 2 mPa*s, at least 4 mPa*s, at least 6 mPa*s, or at least 8 mPa*s. In other variations, the protein-rich water-based yellow peel composition exhibited a viscosity of at least 10 mPa*s. -1 The viscosity is measured at a shear rate of less than or equal to 15 mPa*s, less than or equal to 12 mPa*s, or less than or equal to 10 mPa*s.

[0107] emulsification

[0108] The protein-rich water citrus peel compositions provided herein can be further described based on their emulsifying properties. The emulsifying properties of these protein-rich water citrus peel compositions are suitable for use in food products containing immiscible liquid ingredients, such as non-dairy milk or protein beverages. For example, the mouthfeel of a beverage product can be influenced by the droplet size distribution within the beverage; a narrower distribution (whether unimodal or bimodal) and smaller droplet size provide a smooth texture and a uniform mouthfeel.

[0109] Emulsification can be measured using any suitable technique known in the art. For example, in one variation, the droplet size of the emulsion is analyzed by laser diffraction, following the procedure described in Example 4 below.

[0110] In some embodiments, the emulsion comprising a protein-rich water citrus peel composition may have different droplet size distributions. In some embodiments, the emulsion comprising a protein-rich water citrus peel composition has a unimodal droplet size distribution. In other embodiments, the emulsion comprising a protein-rich water citrus peel composition has a bimodal droplet size distribution.

[0111] In some variations, the emulsion comprising the protein-rich water lily composition has an average droplet size of at least 1 μm, at least 2.5 μm, at least 5 μm, at least 10 μm, at least 25 μm, at least 50 μm, or at least 75 μm. In other variations, the emulsion comprising the protein-rich water lily composition has an average droplet size of less than or equal to 150 μm, less than or equal to 100 μm, less than or equal to 75 μm, or less than or equal to 50 μm. In some variations, the emulsion comprising the protein-rich water lily composition has a bimodal droplet size distribution, wherein the bimodal distribution has a first average droplet size of about 1 μm and a second average droplet size between about 10 μm and about 100 μm.

[0112] In some embodiments, when emulsified, the protein-rich water citrus composition (in emulsion form) has an average droplet size of at least 1 μm, at least 2.5 μm, at least 5 μm, at least 10 μm, at least 25 μm, at least 50, or at least 75 μm. In other embodiments, when emulsified, the protein-rich water citrus composition has an average droplet size of less than or equal to 150 μm, less than or equal to 100 μm, less than or equal to 75 μm, or less than or equal to 50 μm. In some variations, when emulsified, the protein-rich water citrus composition has a bimodal droplet size distribution, wherein the bimodal distribution has a first average droplet size of about 1 μm and a second average droplet size between about 10 μm and about 100 μm.

[0113] In some embodiments, the emulsion of the protein-rich water jasmine composition includes a unimodal droplet size distribution, and the protein-rich water jasmine composition (in emulsion form) can be characterized as having a median droplet size. In other embodiments, when emulsified, the protein-rich water jasmine composition (in emulsion form) has a median droplet size of less than or equal to about 5 μm, less than or equal to about 4 μm, less than or equal to about 35 μm, less than or equal to about 2 μm, or less than or equal to about 1 μm. In some embodiments, when emulsified, the protein-rich water jasmine composition (in emulsion form) has a median droplet size of less than or equal to about 5 μm.

[0114] In some embodiments, the emulsions comprising protein-rich *Phellodendron amurense* compositions provided herein can be further characterized by their stability over time, such as several days after initial preparation. For example, in other embodiments, when emulsified, after storage for 1, 2, 3, 4, 5, 6, or 7 days, the protein-rich *Phellodendron amurense* compositions (in emulsion form) have a median emulsion droplet size of less than or equal to about 5 μm, less than or equal to about 4 μm, less than or equal to about 35 μm, less than or equal to about 2 μm, or less than or equal to about 1 μm. In some embodiments, when emulsified, after storage for 7 days, the protein-rich *Phellodendron amurense* compositions (in emulsion form) have a median droplet size of less than or equal to about 5 μm.

[0115] Foaming properties

[0116] The protein-rich water chestnut compositions provided herein can also be described based on their foaming properties, including their maximum foam production (or foaming capacity) per unit weight of water chestnut composition and their foam stability (e.g., change in foam volume over a specified time period). The foaming properties of the protein-rich water chestnut compositions can be ideal for certain food applications, such as as an egg substitute. The foaming properties were determined according to the scheme described in Example 5 below.

[0117] In some embodiments, the protein-rich *Phellodendron amurense* composition has a foaming capacity of at least about 70 mL, at least about 80 mL, at least about 90 mL, or at least about 100 mL per 60 mL of 0.1% w / v *Phellodendron amurense* protein solution. In some embodiments, the protein-rich *Phellodendron amurense* composition has a foaming capacity of at least about 70 mL per 60 mL of 0.1% w / v *Phellodendron amurense* protein solution. In other embodiments, the protein-rich *Phellodendron amurense* composition has a foaming capacity of less than or equal to about 150 mL, less than or equal to about 140 mL, less than or equal to about 130 mL, less than or equal to about 120 mL, or less than or equal to about 110 mL per 60 mL of 0.1% w / v *Phellodendron amurense* protein solution. In some embodiments, the protein-rich *Phellodendron amurense* composition has a foaming capacity of less than or equal to about 150 mL per 60 mL of 0.1% w / v *Phellodendron amurense* protein solution. In other embodiments, the protein-rich water citrus composition has a foaming capacity between about 70 mL and about 150 mL of 60 mL of 0.1% w / v water citrus protein solution, between about 70 mL and about 120 mL of 60 mL of 0.1% w / v water citrus protein solution, or between about 70 mL and about 100 mL of 60 mL of 0.1% w / v water citrus protein solution. In some embodiments, the protein-rich water citrus composition has a foaming capacity between about 70 mL and about 150 mL of 60 mL of 0.1% w / v water citrus protein solution.

[0118] Alternatively, foaming capacity can be described based on foam volume. In some embodiments, the protein-rich water citrus composition has a foam volume of at least about 100%, at least about 110%, at least about 120%, at least about 130%, at least about 140%, or at least about 150% of the volume of a 0.1% w / v water citrus protein solution. In some embodiments, the protein-rich water citrus composition has a foam volume of at least about 100% of the volume of a 0.1% w / v water citrus protein solution. In other embodiments, the protein-rich water citrus composition has a foam volume of less than or equal to about 200%, less than or equal to about 190%, less than or equal to about 180%, less than or equal to about 170%, less than or equal to about 160%, or less than or equal to about 150% of the volume of a 0.1% w / v water citrus protein solution. In some embodiments, the protein-rich water citrus composition has a foam volume of less than or equal to about 200% of the volume of a 0.1% w / v water citrus protein solution. In other embodiments, the protein-rich water citrus composition has a foam capacity between about 100% and about 200%, between about 100% and about 150%, or between about 150% and about 200% of the volume of a 0.1% w / v water citrus protein solution. In some embodiments, the protein-rich water citrus composition has a foam capacity between about 100% and about 200% of the volume of a 0.1% w / v water citrus protein solution.

[0119] In other embodiments, the protein-rich water citrus compositions can be characterized by their foam stability, for example, by measuring the percentage of foam volume after 5 seconds, 5 minutes, 10 minutes, 15 minutes, or 1 hour relative to the maximum foam volume after initial preparation. In some embodiments, the protein-rich water citrus compositions have a foam stability of at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, or at least about 10%.

[0120] Bulk density

[0121] Protein-rich water chestnut proteins can be further described based on their bulk density. The bulk density or volume density of the protein-rich water chestnut compositions provided herein can indicate the relative ease of use, storage, and / or packaging for both large-scale food processing and consumer applications, such as protein powder mixtures.

[0122] In some embodiments, the protein-rich water lily composition has at least about 0.2 g / cm³. 3 At least approximately 0.25 g / cm³ 3 At least approximately 0.3 g / cm³ 3 At least approximately 0.4 g / cm³ 3 At least approximately 0.5 g / cm³3 At least approximately 0.6 g / cm³ 3 At least approximately 0.7 g / cm³ 3 At least approximately 0.8 g / cm³ 3 At least approximately 0.9 g / cm³ 3 Or at least about 1g / cm 3 The bulk density. In some embodiments, the protein-rich water-wheat peel composition has a bulk density of at least about 0.2 g / cm³. 3 The packing density.

[0123] Water holding capacity and oil holding capacity

[0124] In other embodiments, the protein-rich water citrus compositions provided herein can be characterized by their water-holding capacity and / or oil-holding capacity. Water-holding capacity, also known as water-binding capacity or water-absorbing capacity, is a measure of the total amount of water that can be absorbed per unit weight of the substance (e.g., protein powder or the protein-rich water citrus compositions of this disclosure). In one variation, water-holding capacity is determined according to the scheme described in Example 5 below. Similarly, oil-holding capacity is a measure of the total amount of oil that can be absorbed per unit weight of the substance. In one variation, oil-holding capacity is determined according to the scheme described in Example 5 below.

[0125] The water-holding and oil-holding capacities of the protein-rich wampee compositions presented herein suggest their potential utility and suitability for incorporation into foods in which water and oil retention is desired. For example, in non-dairy yogurts and sourdough, high water-holding capacity can help prevent the separation of liquid (whey) from protein (milk solids) during storage. In another instance, protein-rich wampee compositions have been found for use in meat mimetics, where high water-holding and oil-holding capacities can help replicate certain sensory aspects of animal meat's sensory characteristics (such as juiciness). The water-holding and oil-holding capacities of the protein-rich wampee compositions presented herein reflect the methods used to obtain the compositions, as these properties are influenced by protein solubility, the degree of protein denaturation, and the hydrophobic groups exposed on the proteins.

[0126] In some embodiments, the protein-rich water lily composition has a water-holding capacity of at least about 0.5 g of water, at least about 0.7 g of water, at least about 1 g of water, at least about 1.2 g of water, at least about 1.5 g of water, at least about 2 g of water, at least about 2.5 g of water, at least about 3 g of water, or at least about 3.5 g of water per gram of protein-rich water lily component. In some embodiments, the protein-rich water lily composition has a water-holding capacity of at least about 1.5 g of water per gram of protein-rich water lily component.

[0127] In some embodiments, the protein-rich *Phellodendron amurense* composition has a water-holding capacity of at least about 0.5 g of oil, at least about 0.7 g of oil, at least about 1 g of oil, at least about 1.2 g of oil, at least about 1.5 g of oil, at least about 2.5 g of water, at least about 3 g of water, or at least about 3.5 g of oil per gram of protein-rich *Phellodendron amurense* component. In some embodiments, the protein-rich *Phellodendron amurense* composition has a water-holding capacity of at least about 1.2 g of oil or at least about 1.5 g of oil per gram of protein-rich *Phellodendron amurense* component.

[0128] gelling properties

[0129] In other embodiments, the protein-rich wampee compositions of this disclosure can be characterized by their gelling properties. The ability of these protein-rich wampee compositions to form gels indicates their potential for incorporation into food products where a semi-solid gel structure is desired, such as desserts, (non-dairy) yogurts, non-dairy cheeses, puddings, sauces, dips, and spreads. In one variation, the minimum gelling concentration is determined according to the scheme described in Example 5 below.

[0130] In some embodiments, the protein-rich water jasmine composition of this disclosure has a minimum gel concentration of at least about 5g of protein-rich water jasmine composition per 100g total solution, at least about 6g of protein-rich water jasmine composition per 100g total solution, at least about 7g of protein-rich water jasmine composition per 100g total solution, at least about 8g of protein-rich water jasmine composition per 100g total solution, at least about 9g of protein-rich water jasmine composition per 100g total solution, at least about 10g of protein-rich water jasmine composition per 100g total solution, at least about 11g of protein-rich water jasmine composition per 100g total solution, and at least about 12g of protein-rich water jasmine composition per 100g total solution. In some embodiments, the protein-rich water jasmine composition of this disclosure has a minimum gel concentration of at least about 7g of protein-rich water jasmine composition per 100g total solution. In some other examples, the protein-rich water jasmine composition has a minimum gel concentration of at least about 10g of protein-rich water jasmine composition per 100g total solution.

[0131] Powder dispersibility

[0132] In some embodiments, the protein-rich water citrus peel compositions can be described based on their powder dispersibility. Powder dispersibility, or the ability of powder to break down into particles in water, can indicate the suitability of the dry powder for reconstitution in water, such as for certain beverage products (e.g., milk powder or protein shake powder). For example, in one variant, powder dispersibility is determined according to the scheme described in Example 5 below. In some embodiments, the protein-rich water citrus peel compositions have a powder dispersibility of at least about 10%, at least about 12%, at least about 15%, or at least about 17%. In some embodiments, the protein-rich water citrus peel compositions have a powder dispersibility of at least about 10%.

[0133] Taste and color

[0134] In some embodiments, the protein-rich water jasmine composition described herein has a pure taste characteristic without bitterness. In some embodiments, the protein-rich water jasmine composition has a neutral and / or non-bitter taste. In some embodiments, the concentration of bitter compounds present in the protein-rich water jasmine composition, such as quercetin and / or quercetin dione, is less than 500 ppm. In some embodiments, the concentration of bitter compounds present in the protein-rich water jasmine composition, such as quercetin and / or quercetin dione, is less than 200 ppm. In some variations, the concentration of quercetin and / or quercetin dione present in the protein-rich water jasmine composition is undetectable by methods and techniques known in the art for measuring quercetin and / or quercetin dione.

[0135] In some other embodiments, the protein-rich water citrus peel composition described herein is white or light brown. In some embodiments, the protein-rich water citrus peel composition described herein is white. In some other embodiments, the protein-rich water citrus peel composition is light brown.

[0136] In some embodiments, the protein-rich water citrus composition has:

[0137] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0138] (ii) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0139] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0140] (iv) At pH 7, at least about 35% protein solubility;

[0141] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0142] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0143] (vi) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0144] (vii) Each gram of protein-rich water citrus peel contains at least about 1.5g of oil with oil-holding capacity;

[0145] (viii) A minimum gelling concentration of at least about 10g of protein-rich water yellow peel component per 100g;

[0146] (ix) At least about 10% powder dispersibility;

[0147] (x) Neutral, without bitter taste; or

[0148] (xi) White;

[0149] Or any combination of (i)-(xi).

[0150] In other embodiments, the protein-rich water citrus composition has:

[0151] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0152] (ii) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0153] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0154] (iv) At pH 7, at least about 35% protein solubility;

[0155] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0156] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0157] (vi) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0158] (vii) Each gram of protein-rich water citrus peel contains at least about 1.5g of oil with oil-holding capacity;

[0159] (viii) A minimum gelling concentration of at least about 10g of protein-rich water yellow peel component per 100g;

[0160] (ix) At least about 10% powder dispersibility; or

[0161] (x) Neutral, no bitter taste;

[0162] Or any combination of (i)-(x).

[0163] In other embodiments, the protein-rich water-wampee composition has:

[0164] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0165] (ii) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0166] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0167] (iv) At pH 7, at least about 35% protein solubility;

[0168] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0169] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0170] (vii) Neutral, without bitter taste;

[0171] Or any combination of (i)-(vii).

[0172] In other embodiments, the protein-rich water citrus composition has:

[0173] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0174] (ii) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0175] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0176] (iv) At pH 7, at least about 35% protein solubility;

[0177] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0178] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0179] (vii) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0180] (viii) A minimum gelling concentration of at least approximately 10g of protein-rich water citrus peel per 100g; or

[0181] (ix) Neutral, without bitter taste or

[0182] Or any combination of (i)-(x).

[0183] In some embodiments, the protein-rich water citrus composition has:

[0184] (i) at least about 0.2 g / cm 3 The bulk density;

[0185] (ii) At pH 7, at least about 35% protein solubility;

[0186] (iii) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0187] (iv) Each gram of protein-rich water-rich yellow peel contains at least about 1.5g of oil with oil-holding capacity;

[0188] (v) A minimum gelling concentration of at least approximately 10g of protein-rich water-based loquat extract per 100g; or

[0189] (vi) Neutral, without bitter taste;

[0190] Or any combination of (i)-(vi).

[0191] In other embodiments, the protein-rich water citrus composition has:

[0192] (i) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0193] (ii) A minimum gelling concentration of at least approximately 7g of protein-rich water-based loquat extract per 100g; or

[0194] (iii) Neutral, without bitter taste;

[0195] Or any combination of (i)-(iii).

[0196] Method for producing a protein-rich water-yellow peel composition

[0197] In some aspects, this document provides various methods for producing protein-rich water chestnut compositions (including, for example, water chestnut protein concentrates or isolates). The protein-rich water chestnut compositions described herein are derived from water chestnut beans. In some embodiments, such protein-rich water chestnut compositions are produced from various forms of processed water chestnut meal.

[0198] Water yellow peel meal

[0199] In some embodiments, the water-soaked yellow peel meal has the following characteristics:

[0200] (i) Based on fat content of less than 1%, less than 2.5%, or less than 5% by dry weight; or

[0201] (ii) Bitter compounds less than 1,000 ppm, less than 900 ppm, less than 800 ppm, less than 700 ppm, less than 600 ppm, less than 500 ppm, less than 400 ppm, less than 300 ppm, less than 200 ppm or less than 100 ppm;

[0202] Or both (i) and (ii).

[0203] In some embodiments, the water-soaked yellow peel meal has:

[0204] (i) Based on fat content of less than 0.2%, less than 0.5%, or less than 1% by dry weight; or

[0205] (ii) Bitter compounds less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm or less than 1 ppm.

[0206] Or both (i) and (ii).

[0207] As mentioned above, bitter compounds refer to compounds that are naturally found in water croaker beans and have a bitter taste. In some of the aforementioned variations, the bitter compounds present in water croaker meal may include crocin and / or crocinedione. Therefore, in some of the foregoing embodiments, the water flavour peel meal has: (x) less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm or less than 1 ppm of water flavour diketone; (y) less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm or less than 1 ppm of water flavour diketone; (z) a combination of water flavour and water flavour diketone less than 500 ppm, less than 200 ppm, less than 150 ppm, less than 100 ppm, less than 50 ppm, less than 25 ppm, less than 20 ppm, less than 15 ppm, less than 10 ppm, less than 5 ppm or less than 1 ppm.

[0208] In some variants, the water flavour peel meal is defatted and debittered and contains (i) less than 500 ppm of water flavourin; (ii) less than 500 ppm of water flavoured dione; or (iii) less than 500 ppm of a combination of water flavourin and water flavoured dione. In some variants, the water flavour peel meal is defatted and debittered and contains (i) less than 200 ppm of water flavourin; (ii) less than 200 ppm of water flavoured dione; or (iii) less than 200 ppm of a combination of water flavourin and water flavoured dione.

[0209] The water-rich yellow peel meal used in the methods described herein to produce a protein-rich water-rich yellow peel composition can be produced by various methods and techniques known in the art. Reference Figure 1 Method 100 describes an exemplary method for producing a protein-rich water citrus composition. Method 100 describes dehulling 102, mechanical pressing 104, and grinding 106 of water citrus beans to produce a reduced-fat water citrus meal, which may then undergo solvent extraction 108 to produce a defatted and debittered water citrus meal. This defatted and debittered water citrus meal subsequently undergoes protein extraction 110, followed by protein separation / dispersion 112, neutralization 114, pasteurization 116, and drying (e.g., by spray drying or freeze drying) 118 to produce a protein-rich water citrus composition.

[0210] The shelling in step 102 typically involves passing the water-yellow beans through a shelling machine to loosen the outer shell and separate the two parts. Any suitable technique known in the art can be used to achieve shelling and separation. For example, in some variations, shelling is performed by passing the water-yellow beans through an impact shelling machine and loosening the outer shell from the beans. Other types of shelling equipment, such as abrasive / brushing types, can be used for this purpose. Separation of the beans from the shell can be achieved, for example, by a gravity table or a suction device. In some variations, the shelling step can be omitted.

[0211] Then, in step 104, the beans are mechanically pressed (e.g., cold-pressed), which can typically be done using a press to remove free oil and produce a water-based yellow skin meal with reduced fat content (e.g., 10-14% fat). Cold pressing can be performed using any suitable technique known in the art. For example, various devices such as the Farmet FL-200 press can be used for cold pressing. In some variations, pressing may include passing the hulled beans through a device to produce a meal with reduced free oil and fat content. Mechanically pressing the beans produces a partially defatted bean meal, which, in some variations, retains approximately 30-45% of the original water-based yellow skin oil content.

[0212] Then, in step 106, the meal undergoes grinding to disperse the aggregates and produce a slightly less coarse meal with reduced fat content (e.g., particle size ranging from 0.25 mm to 5.0 mm). Grinding can be performed using any suitable technique known in the art. For example, grinding can be performed using equipment such as a hammer mill, FitzMill, or Quadromill.

[0213] In step 108, the milled, reduced-fat meal can be solvent-extracted to produce a defatted, debittered meal. In some variations, the resulting defatted, debittered water flavone protein meal contains less than 200 ppm of water flavonein and water flavone dione. Solvent extraction typically removes oil and inherent flavonoid compounds, such as water flavonein and water flavone dione. In some variations, solvent extraction may include exposing the reduced-fat meal to a selected group of solvents, such as ethyl acetate, ethanol, hexane, or other organic solvents or any combination thereof. Solvent extraction can be carried out at solvent-to-solid ratios of 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, or any ratio falling within the range of the foregoing. In some variations, extraction may last for 1, 2, 3, 5, 6, 8, or 10 hours, or a duration falling within the range of the foregoing. In some variations, extraction is carried out at temperatures of 25°C, 45°C, 55°C, 60°C, or 65°C, or temperatures falling within any of the aforementioned ranges. In other variations, solvent extraction may be performed in two or more sequential extractions, wherein the solvent used for each extraction may be the same or different. The solvent is then removed, for example by evaporation, to produce a defatted and debittered water-grown yellow skin protein meal rich in protein (e.g., 30-39%) and carbohydrates (e.g., 55-60%).

[0214] In some embodiments, between steps 108 and 110 of exemplary method 100, defatted and debittered water citrus peel meal may be ground to a smaller, more uniform particle size. Any suitable method or technique can be used to grind the meal. For example, in one variation, a coffee grinder or grain mill / flour mill may be used, with or without subsequent sieving. In some embodiments, the defatted and debittered water citrus peel meal is ground to a particle size of less than or equal to 0.5 mm in diameter. In some embodiments, the defatted and debittered water citrus peel meal is uniformly ground to a particle size of less than or equal to 0.5 mm, less than or equal to 0.2 mm, less than or equal to 0.15 mm, or less than or equal to 0.05 mm in diameter. In one embodiment, the defatted and debittered water citrus peel meal is uniformly ground to a particle size of less than or equal to 0.5 mm in diameter.

[0215] Refer again Figure 1 While exemplary method 100 describes the production of a protein-rich water-yellow peel composition from water-yellow peel beans, it should be understood that in other exemplary methods, a protein-rich water-yellow peel composition can be produced from water-yellow peel meal, which can be obtained from any method or technique known in the art or from any commercially available source. In other words, in other embodiments, the protein-rich water-yellow peel composition can be produced from various forms of water-yellow peel meal.

[0216] Similarly, refer to Figure 2A-2CIn exemplary methods 200, 300, and 400, it should be understood that although defatted and debittered water citrus peel meal is used as the starting material, other forms of water citrus peel meal may be used in other variations. For example, in some embodiments, the water citrus peel meal may be milled full-fat water citrus peel meal or reduced-fat water citrus peel meal.

[0217] In some variations, water citrus meal is obtained by dehulling and grinding water citrus beans. In one variation, the ground water citrus meal has: (i) less than or equal to 25% water citrus protein based on dry weight; (ii) at least 30% fat based on dry weight; or (iii) less than or equal to 20,000 ppm of bitter compounds (such as citrus quercetin and / or citrus quercetin dione), or any combination of (i)-(iii).

[0218] In other variations, water citrus meal is a reduced-fat water citrus meal obtained by dehulling, pressing (e.g., cold pressing), and grinding water citrus beans. In one variation, the reduced-fat water citrus meal has: (i) less than or equal to 30% water citrus protein based on dry weight; (ii) less than or equal to 15% fat based on dry weight; or (iii) less than or equal to 10,000 ppm of bitter compounds, or any combination of (i)-(iii).

[0219] In other variations, the water citrus peel meal is defatted and debittered water citrus peel meal obtained by solvent extraction of the reduced-fat water citrus peel meal described above. For example, in some variations, suitable solvents for this solvent extraction may include organic solvents such as esters (e.g., ethyl acetate), alcohols (e.g., methanol, ethanol, etc.), and alkanes (e.g., hexane). In one variation, the defatted and debittered water citrus peel meal has: (i) less than or equal to 40% water citrus peel protein by dry weight; or (ii) less than or equal to 5% fat by dry weight; or (iii) less than or equal to 500 ppm of bitter compounds, or any combination of (i)-(iii). In another variation, the defatted and debittered water citrus peel meal has: (i) less than or equal to 40% water citrus peel protein by dry weight; (ii) less than or equal to 5% fat by dry weight; or (iii) less than or equal to 200 ppm of bitter compounds, or any combination of (i)-(iii).

[0220] In protein extraction step 110, the protein in the defatted and debittered water hyacinth bean cake dissolves into the liquid extract, as discussed in further detail below. Protein separation and separation 112 can be performed using various solid-liquid separation techniques. For example, in some variations, a protein-rich water hyacinth bean composition can be obtained by decantation of the supernatant protein liquid extract (residual solids) from the wet water hyacinth bean cake. In some variations, solubilization can be used to obtain a protein-rich water hyacinth bean composition. In some variations, isoelectric precipitation can be used to obtain a protein-rich water hyacinth bean composition. In some variations, membrane filtration can be used to obtain a protein-rich water hyacinth bean composition.

[0221] Solubilization

[0222] In one aspect, a method is provided for producing a protein-rich water-wheat peel composition by solubilizing water-wheat peel meal. (Reference) Figure 2A In the exemplary method 200, in step 202, an aqueous slurry is prepared using defatted and debittered water citrus peel meal, and the pH is adjusted to an alkaline pH (e.g., between 8 and 10) using a suitable alkali, such as sodium hydroxide. In step 204, water citrus peel protein is extracted by separating the alkaline protein liquid fraction from the insoluble wet filter cake fraction. In other variations of step 202, an aqueous slurry is prepared using defatted and debittered water citrus peel meal, the pH is adjusted to a pH between 6 and 10 using a suitable acid or alkali, and the protein liquid fraction is separated from the insoluble wet filter cake fraction.

[0223] This separation can be achieved using any suitable technique known in the art. For example, it can be achieved using a decanter or a centrifuge. A protein liquid is obtained, and in step 206, the protein liquid is neutralized by adjusting its pH to approximately 7.0, for example, by adding a suitable acid, such as hydrochloric acid or phosphoric acid. In step 208, the neutralized protein liquid is concentrated. In steps 210 and 212, the concentrated protein liquid is subjected to pasteurization and spray drying, respectively, to obtain a protein-rich aqueous wampee composition. The neutralization, concentration, and pasteurization steps are optionally included in the exemplary method.

[0224] In one aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0225] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0226] Adjust the pH of the aqueous slurry to between 8 and 10;

[0227] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0228] Optionally neutralize, concentrate, and / or pasteurize protein liquid fractions; and

[0229] The protein liquid is dried to provide a protein-rich water-based wampee composition.

[0230] In another aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0231] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0232] Adjust the pH of the aqueous slurry to between 6 and 10;

[0233] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0234] Optionally neutralize, concentrate, and / or pasteurize protein liquid fractions; and

[0235] The protein liquid is dried to provide a protein-rich water-based wampee composition.

[0236] In some embodiments, the step of preparing the aqueous slurry may include combining water-based wampee pulp with water. In some embodiments, the method further includes agitating or mixing (e.g., under high shear) the water-based wampee pulp and water.

[0237] As discovered in this disclosure, water citrus peel protein is found to be highly soluble in alkaline aqueous media. Adjusting the aqueous slurry comprising water citrus peel meal to an alkaline pH facilitates the extraction or dissolution of the water citrus peel protein into the solution. In some embodiments, the aqueous slurry is adjusted to a pH between 8 and 10. In other embodiments, the aqueous slurry is adjusted to a pH between 6 and 10.

[0238] Subsequently, in step 204, the slurry is separated into an alkaline protein liquid fraction and an insoluble wet filter cake fraction. This separation can be achieved using solid-liquid separation techniques known in the art, including, for example, decantation and centrifugation.

[0239] In some variations, steps 202 and 204, used for pH adjustment and separation of the protein liquid fraction, can also be performed once or multiple times on the obtained wet filter cake fraction to increase protein yield. That is, as with residual insoluble wet filter cake fractions, the water chestnut meal can be solubilized several times in successive iterations. For example, a wet filter cake can be prepared in a second aqueous slurry and adjusted to an alkaline pH, after which the second alkaline protein liquid fraction is separated from the insoluble filter cake fraction and the two liquid fractions are combined. For repeated extraction, water chestnut meal can be prepared in an aqueous slurry and the pH adjusted to an acidic pH (e.g., pH 2) or an alkaline pH (e.g., pH 8), the resulting pH-adjusted slurry is separated into its protein liquid fraction and insoluble wet filter cake fraction, and a wet filter cake fraction prepared in a subsequent aqueous slurry is then adjusted to an alkaline pH and separated into another protein liquid fraction. Any protein liquid fractions obtained from sequential solubilization are synthesized into a single protein liquid fraction prior to subsequent processing. In other variations, enzymes used to digest carbohydrates in watery yellow peel meal can be added to the aqueous slurry to help increase protein volume.

[0240] In some embodiments, the protein liquid fraction is neutralized to a pH of approximately 7. This neutralization can be achieved by adding a suitable food-grade acid, such as phosphoric acid or hydrochloric acid. In some embodiments, neutralizing the protein liquid fraction involves adding phosphoric acid or hydrochloric acid to the protein liquid fraction.

[0241] In other embodiments, the protein liquid fraction is concentrated after neutralization. Concentration of the protein liquid fraction may involve reducing the liquid volume in the protein liquid fraction to facilitate handling or downstream processing such as acid precipitation and / or membrane filtration.

[0242] In other embodiments, the protein liquid fraction is pasteurized. Pasteurization is a standard food processing technique in which products intended for human consumption are treated with gentle heat, typically below 100°C (212°F), to eliminate pathogens and extend shelf life.

[0243] In a further embodiment, the protein liquid extract is dried to provide a final protein-rich water-based wampee composition. The protein liquid extract can be dried by methods known in the art, including, for example, spray drying and / or lyophilization (e.g., freeze-drying).

[0244] In some variations, the protein-rich *Phellodendron amurense* composition obtained by the aforementioned method is a *Phellodendron amurense* protein concentrate. In some variations, the protein-rich *Phellodendron amurense* composition has at least 40%, at least 50%, or between 50% and 70% *Phellodendron amurense* protein based on dry weight. In other variations, the protein-rich *Phellodendron amurense* composition has at least 40% *Phellodendron amurense* protein based on dry weight; and less than or equal to 40% carbohydrates based on dry weight.

[0245] Immediate precipitation

[0246] In another aspect, this paper provides a method for producing a protein-rich aquatic wampee composition from aquatic wampee meal by precipitation without pre-solubilizing the aquatic wampee protein. Because the aquatic wampee protein is precipitated directly from the aqueous slurry of the aquatic wampee meal without pre-solubilization, this method allows for the recovery of a wider range of proteins from the aquatic wampee meal, including insoluble proteins, requires fewer processing steps overall, increases protein yield, and reduces water and energy consumption. Furthermore, the wider range of protein recovery in combination with carbohydrate content in the compositions obtained by this method can provide enhanced functionality for specific food applications.

[0247] In some embodiments of this aspect, an aqueous slurry is prepared using defatted and debittered water citrus peel meal, and the pH is adjusted to an acidic pH (e.g., between 3 and 5, between 4 and 5, or between pH 4 and 4.5) to induce protein solid precipitation. Upon addition of acid to the slurry, the proteins present in the water citrus peel meal immediately precipitate from the solution. The precipitated proteins separate from the aqueous slurry. This separation can be achieved using any suitable technique known in the art. For example, this separation can be achieved using a decanter or a centrifuge.

[0248] The precipitated protein is washed with water and neutralized with alkali. The neutralized protein is then pasteurized and dried. In the exemplary method described above, the washing, neutralization, concentration, and pasteurization of the protein are optional.

[0249] In one aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0250] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0251] Adjust the pH of the aqueous slurry to between 4 and 5 to obtain water-based yellow skin protein solids;

[0252] Washing, neutralization, and pasteurization purify protein solids; and

[0253] The purified water yellow peel protein solid is dried to provide a protein-rich water yellow peel composition.

[0254] In another aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0255] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0256] The pH of the aqueous slurry was adjusted to acidic pH to obtain water-based yellow peel protein solids;

[0257] Washing, neutralization, and pasteurization purify protein solids; and

[0258] The purified water yellow peel protein solid is dried to provide a protein-rich water yellow peel composition.

[0259] As described above, water citrus protein is precipitated from an aqueous slurry containing water citrus pulp by adjusting the pH of the slurry to an acidic pH, such as between 3 and 5.5. In some embodiments, the pH of the protein liquid is adjusted to a pH between 3 and 5.5, between 3 and 5, between 3 and 4.5, between 3 and 4, between 3 and 3.5, between 3.5 and 5.5, between 3.5 and 5, between 3.5 and 4.5, between 3.5 and 4, between 4 and 5.5, between 4 and 5, between 4 and 4.5, between 4.5 and 5.5, or between 4.5 and 5. In some variations, the pH is adjusted to a pH between 4 and 4.5.

[0260] The water-rich yellow peel protein solids are then separated from the aqueous slurry. Separation can be achieved using solid-liquid separation techniques known in the art, including, for example, decantation and centrifugation.

[0261] In some variations, the method further includes washing the solid purified water to obtain the yellow peel protein.

[0262] In some embodiments, the water citronella protein solid is neutralized to a pH of approximately 7. Purification of the water citronella protein solid can be achieved by neutralizing it with a suitable food-grade alkali, such as sodium hydroxide. In some embodiments, neutralizing the water citronella protein solid involves adding sodium hydroxide to the purified water citronella protein solid.

[0263] In other embodiments, the purified water-based wampee protein solids are concentrated after neutralization. It should be recognized that neutralization of the purified water-based wampee protein solids can result in partial or complete solubilization. Concentration of the purified water-based wampee protein solids may involve reducing the volume of liquid remaining after precipitation or introduced during neutralization by drying or decantation of the purified water-based wampee protein solids, for easier handling or downstream processing.

[0264] In other embodiments, the water-yellow peel protein solid is pasteurized.

[0265] In a further embodiment, the water citrus protein solids are dried to provide a final protein-rich water citrus composition. The water citrus protein solids can be dried by methods known in the art, including, for example, spray drying and / or lyophilization (e.g., freeze-drying).

[0266] In some variations, the protein-rich *Phellodendron amurense* composition obtained by the aforementioned method is a *Phellodendron amurense* protein concentrate. In some variations, the protein-rich *Phellodendron amurense* composition has at least 40% or at least 50% *Phellodendron amurense* protein based on dry weight; or between 40% and 50% or between 40% and 70%. In other variations, the protein-rich *Phellodendron amurense* composition has at least 40% *Phellodendron amurense* protein based on dry weight; and less than or equal to 50% carbohydrates based on dry weight.

[0267] Isoelectric precipitation

[0268] In one aspect, a method is provided for producing a protein-rich water pomegranate composition from water pomegranate meal via isoelectric precipitation. (Reference) Figure 2B In the exemplary method 300, in step 302, an aqueous slurry is prepared using defatted and debittered water citrus peel meal, and the pH is adjusted to between 8 and 10. In step 304, water citrus peel protein is extracted by separating the alkaline protein liquid fraction from the insoluble wet filter cake fraction. This separation can be achieved using any suitable technique known in the art. For example, this separation can be achieved using a decanter or a centrifuge.

[0269] In another variation of step 302, an aqueous slurry is prepared using defatted and debittered water yellow peel meal, and the pH is adjusted to between 6 and 10 with a suitable acid or alkali, and the protein liquid fraction is separated from the insoluble wet filter cake fraction.

[0270] In some variations of step 302, the aqueous slurry can be adjusted to a pH between 6 and 10, and the protein liquid fraction can be separated from the aqueous slurry. A protein liquid is obtained, and in step 306, isoelectric precipitation is used to precipitate a large amount of alkali-soluble water citrus protein. In some variations, isoelectric precipitation is carried out at a pH at or below the isoelectric point of the water citrus protein (e.g., at a pH between 4.0 and 4.5). In step 306, the pH of the alkaline protein liquid fraction is adjusted to a pH between 4.0 and 4.5 by adding an acid such as phosphoric acid or hydrochloric acid. The precipitated water citrus protein solid is collected. In step 308, the precipitated protein is washed. In step 310, the washed protein is neutralized by adjusting the pH to approximately 7.0, for example, with an alkali such as sodium hydroxide. In step 312, the neutralized protein is concentrated. In steps 314 and 316, the concentrated protein liquid is pasteurized and dried (e.g., by spray drying or freeze-drying / lyophilization) to obtain a protein-rich water citrus composition. The washing, neutralization, concentration, and pasteurization of proteins in the exemplary methods described above are optional.

[0271] In one aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0272] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0273] Adjust the pH of the aqueous slurry to between 8 and 10.

[0274] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0275] At least a portion of the water yellow peel protein was precipitated from the liquid protein fraction to obtain purified water yellow peel protein solid;

[0276] Washing, neutralization, and pasteurization purify protein solids; and

[0277] The purified water yellow peel protein solid is dried to provide a protein-rich water yellow peel composition.

[0278] In another aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0279] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0280] Adjust the pH of the aqueous slurry to between 6 and 10.

[0281] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0282] At least a portion of the water yellow peel protein was precipitated from the liquid protein fraction to obtain purified water yellow peel protein solid;

[0283] Washing, neutralization, and pasteurization purify protein solids; and

[0284] The purified water yellow peel protein solid is dried to provide a protein-rich water yellow peel composition.

[0285] As described above, water-rich yellow peel protein is solubilized from aqueous pulp containing water-rich yellow peel meal by adjusting the pH of the pulp to an alkaline pH, for example, between 8 and 10. In other variations, water-rich yellow peel protein is solubilized from aqueous pulp containing water-rich yellow peel meal by adjusting the pH of the pulp to between 6 and 10.

[0286] In step 304, the slurry is then separated into an alkaline protein liquid fraction and an insoluble wet filter cake fraction. This separation can be achieved using solid-liquid separation techniques known in the art, including, for example, decantation and centrifugation.

[0287] In some variations, steps 302 and 304, used for pH adjustment and separation of the protein liquid fraction, can also be performed once or multiple times on the obtained wet filter cake fraction to increase protein yield. That is, as with residual insoluble wet filter cake fractions, the water chestnut meal can be solubilized several times in successive iterations. For example, a wet filter cake can be prepared in a second aqueous slurry and adjusted to an alkaline pH, after which the second alkaline protein liquid fraction is separated from the insoluble filter cake fraction and the two liquid fractions are combined. For repeated extraction, water chestnut meal can be prepared in an aqueous slurry and the pH adjusted to an acidic pH (e.g., pH 2) or an alkaline pH (e.g., pH 8), the resulting pH-adjusted slurry is separated into its protein liquid fraction and insoluble wet filter cake fraction, and a wet filter cake fraction is prepared in a subsequent aqueous slurry, subsequently adjusted to an alkaline pH and separated into another protein liquid fraction. In some variations, the protein liquid fractions obtained from continuous solubilization can be combined into a single protein liquid fraction before subsequent processing. In some variations, solubilization is achieved through countercurrent extraction. In other variations, enzymes for digesting carbohydrates in the water-soluble yellow peel meal (such as carbohydrate enzymes, including cellulase or amylase) can be added to the aqueous slurry to aid in protein solubilization. In some variations where enzymes for digesting carbohydrates are added to the aqueous slurry, the slurry can be heated to a temperature suitable for achieving enzyme activity, such as 37°C.

[0288] The isoelectric precipitation in step 306 can be carried out at a pH at or near the isoelectric point of the water yellow peel protein to provide water yellow peel protein solids from the protein liquid fraction. For example, as Figure 3The results show that the solubility of water citrus protein decreases substantially between pH 3 and pH 5.5. Adding acid to the protein liquid obtained from step 304 to create an acidic pH between 3 and 5.5 precipitates the water citrus protein solid from the solution. In some embodiments, the pH of the protein liquid is adjusted to between 3 and 5.5, between 3 and 5, between 3 and 4.5, between 3 and 4, between 3 and 3.5, between 3.5 and 5.5, between 3.5 and 5, between 3.5 and 4.5, between 3.5 and 4, between 4 and 5.5, between 4 and 5, between 4 and 4.5, between 4.5 and 5.5, or between 4.5 and 5.

[0289] In some variations, the method further includes washing the solid purified water to obtain the yellow peel protein.

[0290] In some embodiments, the water citronella protein solid is neutralized to a pH of approximately 7. The water citronella protein solid can be purified by neutralizing it with the addition of a suitable food-grade alkali, such as sodium hydroxide. In some embodiments, neutralizing the water citronella protein solid involves adding sodium hydroxide to the purified water citronella protein solid.

[0291] In other embodiments, the purified water-based wampee protein solids are concentrated after neutralization. It should be recognized that neutralization of the purified water-based wampee protein solids can result in partial or complete resolubilization. Concentration of the purified water-based wampee protein solids may include reducing the volume of liquid remaining in the water-based wampee protein solids or introduced prior to neutralization by drying or decantation, to facilitate handling or downstream processing.

[0292] In other embodiments, the water-yellow peel protein solid is pasteurized.

[0293] In a further embodiment, the water citrus protein solids are dried to provide a final protein-rich water citrus composition. The water citrus protein solids can be dried by methods known in the art, including, for example, spray drying and / or lyophilization (e.g., freeze-drying).

[0294] In some variations, the protein-rich *Phellodendron amurense* composition obtained by the aforementioned method is a *Phellodendron amurense* protein isolate. In some variations, the protein-rich *Phellodendron amurense* composition has at least 70%, or between 70% and 90%, of *Phellodendron amurense* protein based on dry weight. In other variations, the protein-rich *Phellodendron amurense* composition has at least 70% *Phellodendron amurense* protein based on dry weight; and less than or equal to 20% carbohydrates based on dry weight.

[0295] Membrane filtration

[0296] In another aspect, a method is provided for producing a protein-rich water wampee composition from water wampee meal via membrane filtration. (See reference...) Figure 2C In exemplary method 400, in step 402, an aqueous slurry is prepared using defatted and debittered water citrus peel meal, and the pH is adjusted to between 6 and 10 (e.g., pH 8 and 10). In step 404, water citrus peel protein is extracted by separating the (alkaline) protein liquid fraction from the insoluble wet filter cake fraction. This separation can be achieved using any suitable technique known in the art. For example, this separation can be achieved using a decanter or a centrifuge. A protein liquid is obtained, and in step 406, the protein liquid undergoes membrane filtration to obtain a purified water citrus peel protein retainer. (See again...) Figure 2C In steps 408, 410, 412 and 414, the retained product is washed, neutralized, pasteurized and dried, respectively, to produce a protein-rich water-based wampee composition.

[0297] In one aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0298] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0299] Adjust the pH of the aqueous slurry to between 6 and 10.

[0300] Separating the protein liquid fraction from aqueous slurry,

[0301] The separated protein liquid fraction is passed through a membrane system to obtain a retention including water-rich yellow peel protein;

[0302] Optional washing, neutralization, and / or pasteurization of the residue; and

[0303] The retained material is dried to provide a protein-rich water-based wampee composition.

[0304] In another aspect, this article provides a method for producing a protein-rich water-based wampee composition, comprising:

[0305] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0306] Adjust the pH of the aqueous slurry to between 8 and 10.

[0307] Separating the protein liquid fraction from aqueous slurry,

[0308] The separated protein liquid fraction is passed through a membrane system to obtain a retention including water-rich yellow peel protein;

[0309] Optional washing, neutralization, and / or pasteurization of the residue; and

[0310] The retained material is dried to provide a protein-rich water-based wampee composition.

[0311] As described above, water-rich yellow peel protein is solubilized from an aqueous slurry containing water-rich yellow peel meal. In one aspect, water-rich yellow peel protein is solubilized from the aqueous slurry containing water-rich yellow peel meal at a pH between 6 and 10. In other aspects, water-rich yellow peel protein is solubilized from the aqueous slurry containing water-rich yellow peel meal by adjusting the pH of the slurry to alkaline, for example, between 8 and 10.

[0312] Subsequently, in step 404, the slurry is separated into an alkaline protein liquid fraction and an insoluble wet filter cake fraction. Separation can be achieved using solid-liquid separation techniques known in the art, including, for example, decantation and centrifugation.

[0313] In some variations, steps 402 and 404, used for pH adjustment and separation of the protein liquid fraction, can be performed once or multiple times on the obtained wet filter cake fraction to increase protein yield. That is, as with residual insoluble wet filter cake fractions, the water chestnut meal can be solubilized several times in successive iterations. For example, a wet filter cake can be prepared in a second aqueous slurry and adjusted to an alkaline pH, after which a second alkaline protein liquid fraction is separated from the insoluble filter cake fraction and the two liquid fractions are combined. For repeated extraction, water chestnut meal can be prepared in an aqueous slurry, and the pH can be kept as is or adjusted to an acidic pH (e.g., pH 2) or an alkaline pH (e.g., pH 8). The resulting slurry, either as is or with adjusted pH, is separated into its protein liquid fraction and insoluble wet filter cake fraction, and a wet filter cake fraction is prepared in a subsequent aqueous slurry, subsequently adjusted to an alkaline pH and separated into another protein liquid fraction. Any protein liquid fractions obtained from continuous solubilization can be combined into a single protein liquid fraction prior to subsequent processing. In other variations, enzymes used to digest carbohydrates in watery yellow peel meal can be added to the aqueous slurry to aid in protein dissolution.

[0314] refer to Figure 2CIn step 406, the protein liquid fraction is filtered through a membrane system. Membrane filtration can be performed using different membranes with varying cutoff sizes, variable transmembrane pressures and concentration factors, and percolation factors. In some variations, the protein liquid is passed through a 5 kDa molecular weight cutoff (MWCO) membrane filter or a 10 kDa MWCO membrane filter. In some other variations, the protein liquid is passed through a 5 kDa MWCO membrane filter. In still further variations, the protein liquid is passed through a 5 kDa MWCO membrane filter or a 10 kDa MWCO membrane filter with a concentration factor (CF) of 0-5, 0-4, 0-2, 2-5, 2-4, or 4-5, and a percolation factor (DF) of 0-10, 0-5, 0-4, 0-2, 2-10, 2-5, 2-4, 4-10, or 4-5.

[0315] The protein-rich retentate obtained from membrane filtration can be further optionally washed, neutralized, and / or pasteurized. In some variations, the method further includes washing the retentate.

[0316] In some embodiments, the retainer is neutralized to a pH of approximately pH 7. The retainer can be neutralized by adding a suitable food-grade acid or base, such as phosphoric acid or hydrochloric acid and sodium hydroxide. In some embodiments, neutralizing the retainer involves adding phosphoric acid or hydrochloric acid and / or sodium hydroxide to the retainer.

[0317] In other embodiments, the contents are pasteurized.

[0318] In a further embodiment, the residue is dried to provide a final protein-rich aquamarine composition. The aquamarine protein solids can be dried by methods known in the art, including, for example, spray drying and / or lyophilization (e.g., freeze-drying).

[0319] In some variations, the protein-rich *Phellodendron amurense* composition obtained by the aforementioned method is a *Phellodendron amurense* protein isolate. In some variations, the protein-rich *Phellodendron amurense* composition has at least 70%, or between 70% and 95%, of *Phellodendron amurense* protein based on dry weight. In other variations, the protein-rich *Phellodendron amurense* composition has at least 70% *Phellodendron amurense* protein based on dry weight; and less than or equal to 20% carbohydrates based on dry weight.

[0320] Food, beverages and other products

[0321] In some respects, food and beverage products incorporating or using the protein-rich water citrus composition described herein are also provided. This protein-rich water citrus composition can be used for protein fortification in a variety of food and beverage products, including, for example, fruit juice-based high-acid beverages, allergen-free non-dairy low-acid beverages, plant-based yogurt, plant-based ice cream, baked goods, baked goods, cream soups, meat analogues, and cheese analogues.

[0322] In some embodiments, suitable food products may include, for example, soups, sauces, dressings, hummus, bread, cookies, biscuits, nutrition bars, meal replacement products, and snacks. In some variations, the food incorporating the protein-rich wampee composition described herein, or foods derived therefrom, are baked goods.

[0323] In some implementations, the beverage may include, for example, high-acid beverages, neutral beverages, carbonated beverages, non-carbonated beverages, high-protein beverages, and meal replacement drinks.

[0324] In one aspect, this article provides food products, beverage products, dietary supplements, or other products that include the protein-rich water lily extract provided herein, wherein the protein-rich water lily extract has

[0325] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0326] (ii) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0327] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0328] (iv) At pH 7, at least about 35% protein solubility;

[0329] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0330] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0331] (vi) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0332] (vii) Each gram of protein-rich water citrus peel contains at least about 1.5 grams of oil with oil-holding capacity;

[0333] (viii) A minimum gelling concentration of at least about 10g of protein-rich water yellow peel component per 100g;

[0334] (ix) At least about 10% powder dispersibility; or

[0335] (x) Neutral, no bitter taste;

[0336] Or any combination of (i)-(x).

[0337] As described herein, the properties of the protein-rich aquatic wampee compositions are suitable for use as ingredients in a variety of food applications. In addition to their high protein content, the protein-rich aquatic wampee compositions or ingredients provided herein possess numerous advantageous properties, making them suitable for a wide range of food and beverage products. Regarding certain applications, the protein-rich aquatic wampee ingredients provided herein exhibit superior properties compared to other plant-based protein ingredients on the market (such as peas and soybeans), and therefore can be advantageously incorporated into specific food products relative to competing protein sources. Exemplary products may include, but are not limited to, beverage products (such as ready-to-drink beverages or protein shake powders), dairy substitutes (including plant-based yogurt, cheese, or milk), meat substitute products (such as plant-based burgers), and egg substitutes.

[0338] In other embodiments, this document provides a beverage product comprising a protein-rich water citrus peel ingredient, wherein the protein-rich water citrus peel ingredient has:

[0339] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0340] (ii) Foaming capacity of a 0.1% w / v water-based yellow peel protein solution volume between approximately 100% and approximately 200%;

[0341] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0342] (iv) At pH 7, at least about 35% protein solubility;

[0343] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0344] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0345] (vii) Neutral, without bitter taste;

[0346] Or any combination of (i)-(vii).

[0347] In other embodiments, this document provides a dairy product alternative comprising a protein-rich water citrus component, wherein the protein-rich water citrus component has:

[0348] (i) in 100s -1 The shear rate of the viscosity is between approximately 2 mPa*s and approximately 100 mPa*s.

[0349] (ii) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0350] (iii) At least about 0.2 g / cm³ 3 The bulk density;

[0351] (iv) At pH 7, at least about 35% protein solubility;

[0352] (v) Median emulsion droplet size less than or equal to approximately 5 μm;

[0353] (vi) Median emulsion droplet size less than or equal to approximately 5 μm after 7 days of storage;

[0354] (vii) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0355] (viii) A minimum gelling concentration of at least approximately 10g of protein-rich water citrus peel per 100g; or

[0356] (ix) Neutral, without bitter taste or

[0357] Or any combination of (i)-(x).

[0358] In some embodiments, this document provides meat substitute products comprising a protein-rich water citrus peel ingredient, wherein the protein-rich water citrus peel ingredient has:

[0359] (i) at least about 0.2 g / cm 3 The bulk density;

[0360] (ii) At pH 7, at least about 35% protein solubility;

[0361] (iii) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least about 1.5g of water;

[0362] (iv) Each gram of protein-rich water-rich yellow peel contains at least about 1.5g of oil with oil-holding capacity;

[0363] (v) A minimum gelling concentration of at least approximately 10g of protein-rich water-based loquat extract per 100g; or

[0364] (vi) Neutral, without bitter taste;

[0365] Or any combination of (i)-(vi).

[0366] In other respects, this article provides an egg substitute comprising a protein-rich water wampee component, wherein the protein-rich water wampee component has:

[0367] (i) Foaming capacity of 0.1% w / v protein solution volume between approximately 100% and approximately 200%;

[0368] (ii) A minimum gelling concentration of at least approximately 7g of protein-rich water-based loquat extract per 100g; or

[0369] (iii) Neutral, without bitter taste;

[0370] Or any combination of (i)-(iii).

[0371] In some of the aforementioned variations, each serving of the food or beverage product contains at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 15, or 17 grams of aquamarine protein. In some variations, each serving of the food or beverage product contains between 1 and 20 grams of aquamarine protein. In some variations, at least 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, or 95% of the protein in the food or beverage product by weight is derived from aquamarine protein. In some of the aforementioned variations, the aquamarine protein has a PDCAAS of at least 0.7. In other of the aforementioned variations, the aquamarine protein has a PDCAAS of at least 0.85.

[0372] Food and beverage products may include a variety of other components besides the protein-rich water wampee composition described herein. For example, food and beverage products may include, for instance, water, flour, fats and oils, sweeteners (such as sugar), salt, leavening agents, fruit and vegetable juices, thickeners (such as pectin and other hydrocolloids), defoamers, natural and artificial flavorings, preservatives, and colorings.

[0373] In another aspect, methods for preparing food and / or beverage products are provided. These methods may include one or more of mixing / blending, pasteurization and / or sterilization, baking, fermentation, carbonation, leafing, and packaging.

[0374] In other respects, the protein-rich *Phellodendron amurense* composition described herein can be used or incorporated into pharmaceutical products. In certain variations of the foregoing aspects, the protein-rich *Phellodendron amurense* composition has pharmaceutical-grade purity. In other variations, the protein-rich *Phellodendron amurense* composition has a protein purity of ≥99%.

[0375] In other respects, the protein-rich water lily composition of this article can be used or incorporated into dietary supplement products. In some variations of the foregoing aspects, the protein-rich water lily composition has dietary supplement-grade purity. In other variations, the protein-rich water lily composition has a protein purity of ≥99%.

[0376] In other respects, the protein-rich *Phellodendron amurense* composition described herein can be used or incorporated into cosmetics. In certain variations of the foregoing aspects, the protein-rich *Phellodendron amurense* composition has cosmetic-grade purity. In other variations, the protein-rich *Phellodendron amurense* composition has a protein purity of ≥99%.

[0377] In other respects, the protein-rich water lily composition described herein can be used or incorporated into medicinal foods. In certain variations of the foregoing aspects, the protein-rich water lily composition has medicinal food-grade purity. In other variations, the protein-rich water lily composition has a protein purity of ≥99%.

[0378] In other respects, the protein-rich water celery composition of this article can be used or incorporated into infant formula products. In some variations of the foregoing aspects, the protein-rich water celery composition has infant formula-grade purity. In other variations, the protein-rich water celery composition has a protein purity of ≥99%.

[0379] List of implementation methods

[0380] The embodiments listed below are representative aspects of the present invention.

[0381] 1. A protein-rich water hibiscus composition comprising at least 45% water hibiscus protein based on dry weight.

[0382] 2. The composition according to embodiment 1, wherein the composition has a water-rich yellow skin protein content between 45% and 70% based on dry weight.

[0383] 3. The composition according to Embodiment 1, wherein the composition is a water-yellow peel protein concentrate.

[0384] 4. The composition according to any one of embodiments 1 to 3, wherein the composition is derived from water citrus peel meal, wherein the protein-rich water citrus peel composition has at least 1.25 times more water citrus peel protein than water citrus peel meal.

[0385] 5. The composition according to any one of embodiments 1 to 4, wherein the composition has less than 5% fat based on dry weight.

[0386] 6. The composition according to any one of embodiments 1 to 5, wherein the composition has less than 2% fat based on dry weight.

[0387] 7. The composition according to any one of embodiments 1 to 6, wherein the composition has less than 200 ppm of a naturally occurring bitter compound derived from aquatic wampee.

[0388] 8. The composition according to any one of embodiments 1 to 6, wherein the composition has: (i) less than 200 ppm of hydroflavin; or (ii) less than 200 ppm of hydroflavin dione; or (iii) less than 200 ppm of a combination of hydroflavin and hydroflavin dione.

[0389] 9. The composition according to any one of embodiments 1 to 8, wherein the composition has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, at least 6% phenylalanine, or any combination thereof.

[0390] 10. The composition according to any one of embodiments 1 to 9, wherein at least 50% of the protein present in the composition is soluble in water at a pH of at least 6.

[0391] 11. The composition according to any one of embodiments 1 to 10, wherein the composition has a viscosity of at least 2 mPa*s at a shear rate of 100 s⁻¹.

[0392] 12. The composition according to any one of embodiments 1 to 11, wherein the composition, when emulsified, produces an emulsion having an average droplet size of at least 1 μm.

[0393] 13. The composition according to any one of embodiments 1 to 12, wherein the composition has a protein digestibility-corrected amino acid score of at least 0.7.

[0394] 14. The composition according to any one of embodiments 1 to 13, wherein the composition has an average molecular weight distribution of proteins between 10,000 Daltons and 250,000 Daltons.

[0395] 15. A method for producing a protein-rich water-based wampee composition, comprising:

[0396] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0397] Adjust the pH of the aqueous slurry to between 8 and 10;

[0398] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0399] Neutralizing, concentrating, and / or pasteurizing protein liquid fractions; and

[0400] The protein liquid fraction is dried to provide a protein-rich water-based wampee composition.

[0401] 16. The method according to embodiment 15, wherein the protein-rich water hibiscus composition comprises at least 50% water hibiscus protein based on dry weight.

[0402] 17. The method according to embodiment 15, wherein the protein-rich water huangpi composition is a water huangpi protein concentrate.

[0403] 18. The method according to any one of embodiments 15 to 17, further comprising dehulling and grinding water-yellow-skinned beans to produce water-yellow-skinned bean meal.

[0404] 19. The method according to any one of embodiments 15 to 17, further comprising:

[0405] Dehulling water-yellow-skinned beans to produce dehulled water-yellow-skinned beans; and

[0406] Pressing dehulled water-yellow-skinned beans removes at least a portion of the free oil from the beans to produce water-yellow-skinned meal, which has a reduced fat content.

[0407] 20. The method according to embodiment 19 further includes grinding water-soaked yellow peel meal.

[0408] 21. The method according to any one of embodiments 15 to 17, further comprising:

[0409] Dehull water yellow-skinned beans to produce dehulled water yellow-skinned beans;

[0410] Pressing dehulled water-yellow-skinned soybeans to remove at least a portion of the free oil from the water-yellow-skinned soybeans to produce water-yellow-skinned soybean meal with reduced fat content; and

[0411] The fat-reduced water yellow peel meal is combined with a solvent to produce water yellow peel meal, wherein the water yellow peel meal is defatted and debittered.

[0412] 22. The method according to embodiment 21 further includes grinding the water-based yellow peel meal with reduced fat before combining it with a solvent.

[0413] 23. A protein-rich water-wheat peel composition produced by the method of any one of embodiments 15 to 22.

[0414] 24. A food product, beverage product, dietary supplement product or other product comprising: a protein-rich water citrus composition according to any one of embodiments 1 to 14 and 23.

[0415] 25. The product according to embodiment 24, wherein the product is a baked good, a protein supplement, a protein bar, or a non-dairy beverage.

[0416] 26. The product according to embodiment 24, wherein the product is a medical food, infant formula, cosmetic or pharmaceutical product.

[0417] 27. A protein-rich water hibiscus composition comprising at least 70% water hibiscus protein based on dry weight.

[0418] 28. The composition according to embodiment 27, wherein the composition has a water-rich yellow skin protein content between 70% and 90% based on dry weight.

[0419] 29. The composition according to embodiment 27, wherein the composition is a water-yellow peel protein isolate.

[0420] 30. The composition according to any one of embodiments 27 to 29, wherein the composition is derived from water citrus peel meal, wherein the protein-rich water citrus peel composition has at least 1.25 times more water citrus peel protein than water citrus peel meal.

[0421] 31. The composition according to any one of embodiments 27 to 30, wherein the composition has less than 5% fat based on dry weight.

[0422] 32. The composition according to any one of embodiments 27 to 31, wherein the composition has less than 2% fat based on dry weight.

[0423] 33. The composition according to any one of embodiments 27 to 32, wherein the composition has less than 200 ppm of a naturally occurring bitter compound derived from water-borne yellow peel.

[0424] 34. The composition according to any one of embodiments 27 to 33, wherein the composition has: (i) less than 200 ppm of hydroflavin; or (ii) less than 200 ppm of hydroflavin dione; or (iii) less than 200 ppm of a combination of hydroflavin and hydroflavin dione.

[0425] 35. The composition according to any one of embodiments 27 to 34, wherein the composition has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, at least 6% phenylalanine, or any combination thereof.

[0426] 36. The composition according to any one of embodiments 27 to 35, wherein at least 50% of the protein present in the composition is soluble in water at a pH of at least 6.

[0427] 37. The composition according to any one of embodiments 27 to 36, wherein the composition has a concentration that reacts in 100 seconds. -1 Viscosity at a shear rate of at least 2 mPa*s.

[0428] 38. The composition according to any one of embodiments 27 to 37, wherein the composition, when emulsified, produces an emulsion having an average droplet size of at least 1 μm.

[0429] 39. The composition according to any one of embodiments 27 to 38, wherein the composition has a protein digestibility-corrected amino acid score of at least 0.7.

[0430] 40. The composition according to any one of embodiments 27 to 39, wherein the composition has an average molecular weight distribution of proteins between 10,000 Daltons and 250,000 Daltons.

[0431] 41. A method for producing a protein-rich water-based wampee composition, comprising:

[0432] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0433] Adjust the pH of the aqueous slurry to between 8 and 10.

[0434] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0435] At least a portion of the water-yellow peel protein was precipitated from the liquid protein fraction to obtain purified water-yellow peel protein solid;

[0436] Neutralization and pasteurization of purified water containing yellow peel protein solids; and

[0437] The purified water yellow peel protein solid is dried to provide a protein-rich water yellow peel composition.

[0438] 42. The method according to embodiment 41 further includes washing the water-yellow peel protein solid before drying.

[0439] 43. The method according to embodiment 41 or 42, wherein the precipitation step is performed by isoelectric precipitation.

[0440] 44. The method according to any one of embodiments 41 to 43, wherein the protein-rich water hibiscus composition comprises at least 70% water hibiscus protein based on dry weight.

[0441] 45. The method according to any one of embodiments 41 to 43, wherein the protein-rich water citrus composition is a water citrus protein isolate.

[0442] 46. ​​The method according to any one of embodiments 41 to 45, further comprising dehulling and grinding water-yellow-skinned beans to produce water-yellow-skinned bean meal.

[0443] 47. The method according to any one of embodiments 41 to 45, further comprising:

[0444] Dehulling water-yellow-skinned beans to produce dehulled water-yellow-skinned beans; and

[0445] Pressing dehulled water-yellow-skinned beans removes at least a portion of the free oil from the beans to produce water-yellow-skinned meal, which has a reduced fat content.

[0446] 48. The method according to embodiment 47 further includes grinding water-soaked yellow peel meal.

[0447] 49. The method according to any one of embodiments 41 to 45, further comprising:

[0448] Dehull water yellow-skinned beans to produce dehulled water yellow-skinned beans;

[0449] Pressing dehulled water-yellow-skinned soybeans to remove at least a portion of the free oil from the water-yellow-skinned soybeans to produce water-yellow-skinned soybean meal with reduced fat content; and

[0450] The fat-reduced water yellow peel meal is combined with a solvent to produce water yellow peel meal, wherein the water yellow peel meal is defatted and debittered.

[0451] 50. The method according to embodiment 49 further includes grinding the water-based yellow peel meal with reduced fat before combining it with a solvent.

[0452] 51. A protein-rich water-wheat peel composition produced by the method according to any one of embodiments 41 to 50.

[0453] 52. A food product, beverage product, dietary supplement product or other product comprising: a protein-rich water-wampee composition according to any one of embodiments 27 to 40 and 51.

[0454] 53. The product according to embodiment 52, wherein the product is a baked good, a protein supplement, a protein bar, or a non-dairy beverage.

[0455] 54. The product according to embodiment 52, wherein the product is a medical food, infant formula, cosmetic or pharmaceutical product.

[0456] 55. A protein-rich composition of *Phellodendron amurense*, comprising at least 70% *Phellodendron amurense* protein based on dry weight.

[0457] 56. The composition according to embodiment 55, wherein the composition has a water-rich yellow skin protein content between 70% and 90% based on dry weight.

[0458] 57. The composition according to embodiment 55, wherein the composition is a water-yellow peel protein isolate.

[0459] 58. The composition according to any one of embodiments 55 to 57, wherein the composition is derived from water citrus peel meal, wherein the protein-rich water citrus peel composition has at least 1.25 times more water citrus peel protein than water citrus peel meal.

[0460] 59. The composition according to any one of embodiments 55 to 58, wherein the composition has less than 5% fat based on dry weight.

[0461] 60. The composition according to any one of embodiments 55 to 59, wherein the composition has less than 2% fat based on dry weight.

[0462] 61. The composition according to any one of embodiments 55 to 60, wherein the composition has less than 200 ppm of a naturally occurring bitter compound derived from water-borne yellow peel.

[0463] 62. The composition according to any one of embodiments 55 to 61, wherein the composition has: (i) less than 200 ppm of hydroflavin; or (ii) less than 200 ppm of hydroflavin dione; or (iii) less than 200 ppm of a combination of hydroflavin and hydroflavin dione.

[0464] 63. The composition according to any one of embodiments 55 to 62, wherein the composition has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, at least 6% phenylalanine, or any combination thereof.

[0465] 64. The composition according to any one of embodiments 55 to 63, wherein at least 50% of the protein present in the composition is soluble in water at a pH of at least 6.

[0466] 65. The composition according to any one of embodiments 55 to 64, wherein the composition has a concentration that reacts in 100 seconds. -1 Viscosity at a shear rate of at least 2 mPa*s.

[0467] 66. The composition according to any one of embodiments 55 to 65, wherein the composition, when emulsified, produces an emulsion having an average droplet size of at least 1 μm.

[0468] 67. The composition according to any one of embodiments 55 to 66, wherein the composition has a protein digestibility-corrected amino acid score of at least 0.7.

[0469] 68. The composition according to any one of embodiments 55 to 67, wherein the composition has an average molecular weight distribution of proteins between 10,000 Daltons and 250,000 Daltons.

[0470] 69. A method for producing a protein-rich water-based wampee composition, comprising:

[0471] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0472] Adjust the pH of the aqueous slurry to between 8 and 10.

[0473] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0474] The liquid fraction of protein is passed through a membrane system to obtain a retention product including water-rich yellow peel protein;

[0475] Optional washing, neutralization, and / or pasteurization of the residue; and

[0476] The dried residue is used to provide a protein-rich watery wampee composition.

[0477] 70. The method according to embodiment 69, wherein the protein-rich water hibiscus composition comprises at least 70% water hibiscus protein based on dry weight.

[0478] 71. The method according to embodiment 69, wherein the protein-rich water hibiscus composition is a water hibiscus protein isolate.

[0479] 72. The method according to any one of embodiments 69 to 71 further includes dehulling and grinding water-yellow-skinned beans to produce water-yellow-skinned bean meal.

[0480] 73. The method according to any one of embodiments 69 to 71, further comprising:

[0481] Dehulling water-yellow-skinned beans to produce dehulled water-yellow-skinned beans; and

[0482] Pressing dehulled water-yellow-skinned beans removes at least a portion of the free oil from the beans to produce water-yellow-skinned meal, which has a reduced fat content.

[0483] 74. The method according to embodiment 73 further includes grinding water-soaked yellow peel meal.

[0484] 75. The method according to any one of embodiments 69 to 71, further comprising:

[0485] Dehull water yellow-skinned beans to produce dehulled water yellow-skinned beans;

[0486] Pressing dehulled water-yellow-skinned soybeans to remove at least a portion of the free oil from the water-yellow-skinned soybeans to produce water-yellow-skinned soybean meal with reduced fat content; and

[0487] The fat-reduced water yellow peel meal is combined with a solvent to produce water yellow peel meal, wherein the water yellow peel meal is defatted and debittered.

[0488] 76. The method according to embodiment 75 further includes grinding the water-based yellow peel meal with reduced fat before combining it with a solvent.

[0489] 77. A protein-rich water-wampee composition produced by the method according to any one of embodiments 69 to 76.

[0490] 78. A food product, beverage product, dietary supplement product or other product comprising: a protein-rich water-wampee composition according to any one of embodiments 55 to 68 and 77.

[0491] 79. The product according to embodiment 78, wherein the product is a baked good, a protein supplement, a protein bar, or a non-dairy beverage.

[0492] 80. The product according to embodiment 78, wherein the product is a medical food, infant formula, cosmetic or pharmaceutical product.

[0493] 81. A protein-rich component of *Phellodendron amurense*, comprising at least 40% *Phellodendron amurense* protein based on dry weight.

[0494] The components include: (i) less than 500 ppm of hydroflavin; or (ii) less than 500 ppm of hydroflavin dione; or (iii) a combination of hydroflavin and hydroflavin dione less than 500 ppm; and

[0495] The components include carbohydrates with a dry weight of less than 40%.

[0496] 82. The composition according to embodiment 81, wherein the composition has a water-rich yellow peel protein content between 40% and 70% based on dry weight.

[0497] 83. The composition according to embodiment 81, wherein the composition is a water-rich yellow peel protein concentrate.

[0498] 84. The ingredients according to embodiment 81, comprising at least 70% water-based yellow peel protein based on dry weight.

[0499] The components include: (i) less than 500 ppm of hydroflavin; or (ii) less than 500 ppm of hydroflavin dione; or (iii) a combination of hydroflavin and hydroflavin dione less than 500 ppm; and

[0500] The components contain carbohydrates of less than or equal to about 20% by dry weight.

[0501] 85. The composition according to embodiment 84, wherein the composition has a water-rich yellow peel protein content between 70% and 90% based on dry weight.

[0502] 86. The composition according to embodiment 85, wherein the composition is a water-rich yellow peel protein isolate.

[0503] 87. A protein-rich component of *Phellodendron amurense*, comprising at least 40% *Phellodendron amurense* protein based on dry weight.

[0504] The components include: (i) less than 500 ppm of hydroflavin; or (ii) less than 500 ppm of hydroflavin dione; or (iii) a combination of hydroflavin and hydroflavin dione less than 500 ppm; and

[0505] The components include carbohydrates, which make up less than 50% of the dry weight.

[0506] 88. The ingredient according to any one of embodiments 81 to 87, wherein the ingredient is derived from water citrus peel meal, wherein the protein-rich water citrus peel ingredient has a water citrus peel protein content of at least 1.25 times that of water citrus peel meal.

[0507] 89. The ingredient according to any one of embodiments 81 to 88, wherein the ingredient has less than 5% fat based on dry weight.

[0508] 90. The ingredient according to any one of embodiments 81 to 89, wherein the ingredient has less than 2% fat based on dry weight.

[0509] 91. The component according to any one of embodiments 81 to 90, wherein the component has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, at least 6% phenylalanine, or any combination thereof.

[0510] 92. The ingredient according to any one of embodiments 81 to 91, wherein at least 35% of the protein present in the ingredient is soluble in water at a pH of at least 6.

[0511] 93. The component according to any one of embodiments 81 to 92, wherein the component has a concentration of 100s -1 The viscosity with a shear rate of at least 2 mPa*s.

[0512] 94. The component according to any one of embodiments 81 to 93, wherein the component, when emulsified, produces an emulsion having an average droplet size of at least 1 μm.

[0513] 95. The ingredient according to any one of embodiments 81 to 94, wherein the ingredient has a protein digestibility-corrected amino acid score of at least 0.7.

[0514] 96. The component according to any one of embodiments 81 to 95, wherein the component has an average molecular weight of protein between 10,000 Daltons and 250,000 Daltons.

[0515] 97. The composition according to any one of embodiments 81 to 96, wherein the composition includes seed storage protein, and 30-40% of the protein present therein is a protein with a molecular weight between 45 kDa and about 70 kDa as determined by SDS-PAGE.

[0516] 98. The composition according to embodiment 97, wherein the composition further comprises a seed storage protein having a molecular weight of 170-250 kDa, 115-160 kDa, 45-70 kDa, 19-25 kDa, 14-17 kDa, or 10-13 kDa, or any combination thereof.

[0517] 99. The component according to any one of embodiments 81 to 98, wherein the component comprises:

[0518] (i) in 100s -1 The shear rate of viscosity between 2 mPa*s and 100 mPa*s;

[0519] (ii) Foaming ability of 0.1% protein solution volume between 100% and 200%;

[0520] (iii) At least 0.2 g / cm 3 The bulk density;

[0521] (iv) At pH 7, at least 35% protein solubility;

[0522] (v) Median emulsion droplet size less than or equal to 5 μm;

[0523] (vi) Median emulsion droplet size less than or equal to 5 μm after 7 days of storage;

[0524] (vi) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least 1.5g of water;

[0525] (vii) Each gram of protein-rich water citrus peel contains at least 1.5g of oil with oil-holding capacity;

[0526] (viii) A minimum gelling concentration of at least 10g of protein-rich water-based yellow peel components per 100g;

[0527] (ix) At least 10% powder dispersibility; or

[0528] (x) Neutral, no bitter taste;

[0529] Or any combination of (i)-(x).

[0530] 100. The composition according to embodiment 99, wherein the composition comprises:

[0531] (i) in 100s -1 The shear rate of viscosity between 2 mPa*s and 100 mPa*s;

[0532] (ii) Foaming ability of 0.1% w / v water-based yellow peel protein solution volume between 100% and 200%;

[0533] (iii) At least 0.2 g / cm 3 The bulk density;

[0534] (iv) At pH 7, at least 35% protein solubility;

[0535] (v) Median emulsion droplet size less than or equal to 5 μm;

[0536] (vi) Median emulsion droplet size less than or equal to 5 μm after 7 days of storage;

[0537] (vii) Neutral, without bitter taste;

[0538] Or any combination of (i)-(vii).

[0539] 101. The component according to embodiment 99, wherein the component comprises:

[0540] (i) in 100s -1 The shear rate of viscosity between 2 mPa*s and 100 mPa*s;

[0541] (ii) Foaming capacity of 0.1% w / v protein solution volume between 100% and 200%;

[0542] (iii) At least 0.2 g / cm 3 The bulk density;

[0543] (iv) At pH 7, at least 35% protein solubility;

[0544] (v) Median emulsion droplet size less than or equal to 5 μm;

[0545] (vi) Median emulsion droplet size less than or equal to 5 μm after 7 days of storage;

[0546] (vii) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least 1.5g of water;

[0547] (viii) A minimum gelling concentration of at least 10g of protein-rich water-based loquat extract per 100g; or

[0548] (ix) Neutral, without bitter taste or

[0549] Or any combination of (i)-(x).

[0550] 102. The composition according to embodiment 99, wherein the composition comprises:

[0551] (i) at least 0.2 g / cm 3 The bulk density;

[0552] (ii) At pH 7, at least 35% protein solubility;

[0553] (iii) Each gram of protein-rich water-containing yellow peel has a water-holding capacity of at least 1.5g of water;

[0554] (iv) Each gram of protein-rich water-rich yellow peel contains at least 1.5g of oil with oil-holding capacity;

[0555] (v) A minimum gelling concentration of at least 10g of protein-rich water-based loquat extract per 100g; or

[0556] (vi) Neutral, without bitter taste;

[0557] Or any combination of (i)-(vi).

[0558] 103. The composition according to embodiment 99, wherein the composition comprises:

[0559] (i) Foaming capacity of 0.1% w / v protein solution volume between 100% and 200%;

[0560] (ii) A minimum gelling concentration of at least 7g of protein-rich water-based loquat extract per 100g; or

[0561] (iii) Neutral, without bitter taste;

[0562] Or any combination of (i)-(iii).

[0563] 104. A method for producing a protein-rich water-based wampee composition, comprising:

[0564] An aqueous slurry for preparing aquatic citrus peel meal, wherein the aquatic citrus peel meal is defatted and debittered and has (i) less than 500 ppm of aquatic citrus peelin; or (ii) less than 500 ppm of aquatic citrus peel dione; or (iii) less than 500 ppm of a combination of aquatic citrus peelin and aquatic citrus peel dione.

[0565] Adjust the pH of the aqueous slurry to between 6 and 10;

[0566] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0567] Neutralizing, concentrating, and / or pasteurizing protein liquid fractions; and

[0568] The protein liquid fraction is dried to provide a protein-rich water-based wampee composition.

[0569] 105. The method according to embodiment 104, wherein the protein-rich water hibiscus composition comprises at least 50% water hibiscus protein based on dry weight.

[0570] 106. A method for producing a protein-rich water-based component of yellow peel, comprising:

[0571] An aqueous slurry for preparing aquatic citrus peel meal, wherein the aquatic citrus peel meal is defatted and debittered and has (i) less than 500 ppm of aquatic citrus peelin; or (ii) less than 500 ppm of aquatic citrus peel dione; or (iii) less than 500 ppm of a combination of aquatic citrus peelin and aquatic citrus peel dione.

[0572] Adjust the pH of the aqueous slurry to between 6 and 10.

[0573] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0574] At least a portion of the water-yellow peel protein was precipitated from the liquid protein fraction to obtain purified water-yellow peel protein solid;

[0575] Neutralization and pasteurization of purified water containing yellow peel protein solids; and

[0576] The purified water yellow peel protein solids are dried to provide a water yellow peel component rich in protein.

[0577] 107. The method according to embodiment 106 further includes washing the water-yellow peel protein solid before drying.

[0578] 108. The method according to embodiment 106 or 107, wherein the precipitation step is performed by isoelectric precipitation.

[0579] 109. The method according to any one of embodiments 106 to 108, wherein the protein-rich water yellow peel component comprises water yellow peel protein at least 70% by dry weight.

[0580] 110. The method according to any one of embodiments 106 to 109, further comprising:

[0581] Dehulling water-yellow-skinned beans to produce dehulled water-yellow-skinned beans; and

[0582] Pressing dehulled water-yellow-skinned beans removes at least a portion of the free oil from the beans to produce water-yellow-skinned meal, which has a reduced fat content.

[0583] 111. The method according to embodiment 110 further includes grinding water-processed yellow peel meal.

[0584] 112. The method according to any one of embodiments 106 to 111, further comprising:

[0585] Dehull water yellow-skinned beans to produce dehulled water yellow-skinned beans;

[0586] Pressing dehulled water-yellow-skinned soybeans to remove at least a portion of the free oil from the water-yellow-skinned soybeans to produce water-yellow-skinned soybean meal with reduced fat content; and

[0587] The fat-reduced water yellow peel meal is combined with a solvent to produce water yellow peel meal, wherein the water yellow peel meal is defatted and debittered.

[0588] 113. The method according to embodiment 112 further includes grinding the water-based yellow peel meal with reduced fat before combining it with a solvent.

[0589] 114. The method according to embodiment 112 or embodiment 113, wherein the solvent includes ethyl acetate, ethanol, or a combination thereof.

[0590] 115. A method for producing a protein-rich water-yellow peel component, comprising:

[0591] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0592] Adjust the pH of the aqueous slurry to between 6 and 10.

[0593] The slurry was separated into a protein liquid fraction and an insoluble wet filter cake fraction.

[0594] The liquid fraction of protein is passed through a membrane system to obtain a retention product including water-rich yellow peel protein;

[0595] Optional washing, neutralization, and / or pasteurization of the residue; and

[0596] The preserved material is dried to provide a protein-rich watery yellow peel component.

[0597] 116. The method according to embodiment 115, wherein the protein-rich water yellow peel component comprises at least 70% water yellow peel protein based on dry weight.

[0598] 117. The method according to embodiment 115 or embodiment 116, further comprising:

[0599] Dehulling water-yellow-skinned beans to produce dehulled water-yellow-skinned beans; and

[0600] Pressing dehulled water-yellow-skinned beans removes at least a portion of the free oil from the beans to produce water-yellow-skinned meal, which has a reduced fat content.

[0601] 118. The method according to embodiment 117 further includes grinding water-processed yellow peel meal.

[0602] 119. The method according to any one of embodiments 115 to 117, further comprising:

[0603] Dehull water yellow-skinned beans to produce dehulled water yellow-skinned beans;

[0604] Pressing dehulled water-yellow-skinned soybeans to remove at least a portion of the free oil from the water-yellow-skinned soybeans to produce water-yellow-skinned soybean meal with reduced fat content; and

[0605] The fat-reduced water yellow peel meal is combined with a solvent to produce water yellow peel meal, wherein the water yellow peel meal is defatted and debittered.

[0606] 120. The method according to embodiment 119 further includes grinding water-based yellow peel meal with reduced fat before combining with a solvent.

[0607] 121. A method for producing a protein-rich water-based wampee composition, comprising:

[0608] Preparation of an aqueous slurry from water-rich yellow peel meal;

[0609] The pH of the aqueous slurry was adjusted to between 4 and 5 to obtain water-based yellow peel protein solids;

[0610] Washing, neutralization, and pasteurization purify protein solids; and

[0611] The purified water yellow peel protein solid is dried to provide a protein-rich water yellow peel composition.

[0612] 122. A protein-rich water-yellow peel component produced by the method according to any one of embodiments 103 to 121.

[0613] 123. A food product, beverage product, dietary supplement product or other product comprising: the protein-rich water citrus peel ingredient as described in any one of embodiments 81 to 103 and 122.

[0614] 124. The product according to embodiment 123, wherein the product is a baked good, a protein supplement, a protein bar, or a non-dairy beverage.

[0615] 125. The product according to embodiment 123, wherein the product is a medical food, infant formula, cosmetic or pharmaceutical product.

[0616] 126. The product according to embodiment 123, wherein the product is a beverage product, a dairy substitute, a meat substitute product, or an egg substitute.

[0617] 127. The product according to embodiment 126, wherein the beverage product is a fruit smoothie, a meal replacement beverage, a protein drink, or an instant milkshake.

[0618] 128. The product according to any one of embodiments 123, 126 and 127, wherein the product is a beverage product comprising at least 20g of protein-rich water citrus peel ingredient per serving.

[0619] 129. The product according to embodiment 126, wherein the dairy substitutes are non-dairy milk, non-dairy cheese, non-dairy coffee whitening agent or creamer, non-dairy yogurt, non-dairy Greek yogurt, or non-dairy drinking yogurt.

[0620] Example

[0621] The subject matter of this disclosure will be better understood by referring to the following embodiments, which are provided as examples of the invention and not by way of limitation.

[0622] Example 1

[0623] Preparation of defatted and debittered water yellow peel meal

[0624] This embodiment demonstrates the preparation of defatted and debittered water-processed yellow peel meal, and generally follows... Figure 1The method described herein involves pressing water citrus beans through an oil press to remove free oil, thereby producing a reduced-fat water citrus meal with approximately 15-25% fat. The reduced-fat water citrus meal is then extracted with ethanol (5:1 solvent:solvent ratio) at 50-65°C for 3 hours using an oil extractor. Residual solvent is removed by drying. Various determinations are performed on the solvent-extracted meal to analyze various components of the water citrus meal (e.g., moisture, crude fat, protein, carbohydrates, ash, fiber, amino acids, sugars, etc.). The determinations used in this disclosure to evaluate the water citrus meal are derived from AOAC international analytical methods. A brief summary of the determinations is provided and is shown in Table 1 below.

[0625] Table 1. Similarity analysis and amino acid analysis of Phellodendron amurense peel meal and protein-rich Phellodendron amurense peel composition.

[0626]

[0627] Similarity analysis was performed as follows: Total protein content was determined by placing the water-wheat peel meal sample in the combustion chamber of a protein analyzer, measuring the total nitrogen content of the gases produced by combustion, and calculating the protein content from the observed nitrogen content (protein content = 6.25 × nitrogen content). Total fat content was determined by solvent extraction with petroleum ether under reflux (AOCS BA3-38 reference method, modified).

[0628] Total carbohydrate content was calculated as the remaining percentage (100%) of water-wheat peel meal minus the sum of total ash content (%), total protein content (%), total moisture content (%), and total fat content (%). Total ash content was determined by placing a 2g sample of water-wheat peel meal in a crucible, drying the sample in an oven, ashing the sample in a muffle furnace at 600°C, and measuring the weight of the ash (AOAC 942.05 reference method). Total moisture content was determined by heating the weighed sample in a forced-ventilation oven at 130°C for 2 hours and determining the difference in sample weight, where the percentage difference was calculated as moisture content (AOCS BA 2A-38 reference method).

[0629] The contents of quercetin and quercetin dione in the samples of *Phellodendron chinense* were determined by solvent extraction of quercetin and quercetin dione from the samples, followed by HPLC analysis as described in this article.

[0630] The crude fat, protein, hydroflavin, and hydroflavin dione content of the solvent-extracted meal were analyzed. The results are shown in Table 2. The defatted, debittered hydroflavin meal was observed to have less than 0.5% fat and less than approximately 10 ppm hydroflavin and hydroflavin dione levels. The treated meal was observed to have no off-odor and no bitterness.

[0631] Table 2. Approximate Composition of Various Forms of Water-Soybean Meal Raw Materials

[0632]

[0633] Example 2A

[0634] A protein concentrate of water-yellow peel was prepared by solubilization.

[0635] This embodiment demonstrates the extraction of protein from defatted and debittered water citrus peel meal and the production of water citrus peel protein concentrate, and generally follows... Figure 2A The exemplary method described herein.

[0636] Using ethyl acetate as a solvent, defatted and debittered water citrus peel meal was obtained according to the method described in Example 1 above. The ethyl acetate-extracted water citrus peel meal was used as the starting material for protein extraction in this example. An aqueous slurry of the defatted and debittered water citrus peel meal was prepared using water (1:6; 15% solids) in a high-shear mixer. The pH of the slurry was adjusted to pH 8 with NaOH (10M NaOH(aq), about 40% aqueous solution) and continuously stirred at 25°C for 2 hours. The slurry was separated into a protein-containing liquid phase and a wet filter cake by centrifugation. The pH of the protein solution was adjusted to neutral pH (7.0) and freeze-dried to produce water citrus peel protein concentrate. This method resulted in the extraction and recovery of 70-75% total protein by weight from the meal. The water citrus peel protein concentrate contained 50% protein by weight.

[0637] Protein content was determined by total nitrogen using a general conversion factor of 6.25. Nitrogen content was determined using the combustion analysis method described in Example 1 above.

[0638] Tables 3, 4, and 5 below provide the approximate composition, relative amino acid profile, and protein digestibility corrected amino acid score (PDCAAS) of the aquamarine protein concentrate. The approximate composition and amino acid profile were determined using the method described in Example 1 above. The PDCAAS was calculated using a reference amino acid pattern from breast milk as the reference protein. It was observed that the aquamarine protein concentrate produced in this example possessed similar characteristics to soybean protein (USDA Food Data Center database, soybean protein concentrate produced by acid washing (item 16420)) and pea protein (PURIS). TM The amino acid profile is similar and comparable to that of pea protein 870.

[0639] Table 3. Composition of water-wampee protein concentrate, dry weight (excluding moisture).

[0640] parameter ％ Moisture 4.35 Crude fat 0.99 Crude protein (based on dryness) 51.42 Total carbohydrates 39.16 crude fiber 0.30 Total sugar 24.58 sucrose 22.15 fructose 2.43 glucose <0.16 lactose <0.16 maltose <0.16 Ash 8.42

[0641] Table 4. Relative amino acid profiles – proteins from water citrus, soybean, and pea.

[0642]

[0643]

[0644] Note: * indicates essential amino acids.

[0645] Table 5. Amino acid score of water croton protein concentrate

[0646]

[0647] Restricting amino acid = threonine

[0648] Amino acid restriction score = 99

[0649] Digestibility factor = 0.87; PDCAAS = 0.87

[0650] Example 2B

[0651] Protein isolates from water-wampee were prepared by isoelectric precipitation.

[0652] This embodiment demonstrates the extraction of protein from defatted and debittered water citrus peel meal and the production of a protein-rich water citrus peel composition (water citrus peel protein isolate), and generally follows... Figure 2B The exemplary method described herein.

[0653] Method A

[0654] In this embodiment, defatted and debittered water citrus peel meal was used as the starting material for protein extraction. Except for using ethyl acetate as a solvent, the defatted and debittered water citrus peel meal was obtained according to the method described in Example 1 above.

[0655] Aqueous slurry of defatted and debittered water citrus peel meal was prepared using water (1:6; 15% solids) in a high-shear mixer. The pH of the slurry was adjusted to pH 8 with 10M NaOH and continuously stirred at 25°C for 2 hours. The slurry was separated into a protein-containing liquid phase and a wet filter cake by centrifugation. The pH of the protein solution was adjusted to pH 4.5 with phosphoric acid (85% aqueous solution) and stirred for 30 minutes to form a protein precipitate. The precipitated protein was collected by centrifugation, resuspended in water to 40% solids, adjusted to pH 7.0 with 1M NaOH, and freeze-dried into a protein isolate powder. This method resulted in the extraction and recovery of approximately 70-75% by weight of total protein and approximately 40-50% by weight from the defatted and debittered water citrus peel meal. The water citrus peel protein isolate contained approximately 70% by weight of protein. Approximately 38% of the total protein in the starting material was recovered. The approximate composition and amino acid profile of the water citrus peel protein isolate were determined according to the scheme described in Example 1 above. Table 6 shows the moisture, crude fat, protein, carbohydrate, and ash content of the protein isolate from *Phellodendron amurense*. Table 7 provides the relative amino acid profile of the protein isolate from *Phellodendron amurense*.

[0656] Table 6. Composition of protein isolate from water-wampee peel, dry weight (excluding water content)

[0657] parameter ％ Moisture-forced ventilation oven 5.96 Crude fat extracted by petroleum ether <0.11 Protein-burning 76.70 Carbohydrates, calculated 14.08 Ash 9.22

[0658] A watery yellow peel protein isolate was prepared from the meal extracted from ethyl acetate by isoelectric precipitation.

[0659] Table 7. Relative amino acid profiles of protein isolates from *Phellodendron amurense*.

[0660] amino acids g / 100g protein glutamic acid 16.26 Aspartic acid 12.26 Leucine* 9.93 Lysine (total)* 8.69 Phenylalanine* 6.61 Arginine 5.66 Serine 6.03 proline 5.26 Valine* 4.94 glycine 3.70 alanine 3.67 Tyrosine* 3.95 Isoleucine* 3.68 Threonine* 3.22 Histidine* 2.61 Cysteine* 1.31 Tryptophan* 1.27 Methionine* 0.96

[0661] Note: * indicates essential amino acids.

[0662] Method B

[0663] In this embodiment, defatted and debittered water yellow peel meal was used as the starting material for protein extraction. Ethanol was used as the solvent, and the defatted and debittered water yellow peel meal was obtained according to the method described in Example 1 above.

[0664] Aqueous slurry of defatted and debittered water-based yellow peel meal was prepared using water (1:6; 15% solids) in a high-shear mixer. The pH of the slurry was adjusted to pH 8 with 10% NaOH and continuously agitated at 25°C for 1 hour. The slurry was separated into a protein-containing liquid phase and a wet filter cake using a decanter centrifuge. The wet filter cake was resuspended in water, adjusted to pH 8, and agitated at 25°C for another hour. The slurry was again separated into a protein-containing liquid phase and a wet filter cake using a decanter centrifuge. The two protein-containing liquid phases were combined and adjusted to pH 4.5 with phosphoric acid (85% aqueous solution) and agitated for 30 minutes to form a protein precipitate. The precipitated protein was collected by centrifugation, washed with water, resuspended in water (~16% solids), adjusted to pH 7.0 with 10% NaOH, pasteurized, and finally spray-dried into a protein isolate powder.

[0665] During the first extraction, approximately 45% of the total protein present in the starting meal was extracted. An additional 10-14% of the starting protein was recovered by washing the insoluble material (wet filter cake) from the first extraction, resulting in a combined total of 54-59% of the extracted starting protein. Approximately 35-47% of the extracted protein was recovered during the acid precipitation step, resulting in a total yield of 15-26% by weight of the total protein from the defatted and debittered water yellow skin meal.

[0666] The above method was performed twice to obtain two samples of *Phellodendron amurense* protein isolates. Both isolates contained approximately 80% protein by weight. The approximate composition and amino acid profile of the *Phellodendron amurense* protein isolates were determined according to the protocol described in Example 1 above. Table 8 shows the moisture, crude fat, protein, carbohydrate, and ash content of the *Phellodendron amurense* protein isolates. Table 9 provides the relative amino acid profiles of the *Phellodendron amurense* protein isolates.

[0667] Table 8. Composition of protein isolate from water-rich yellow peel, dry weight (excluding moisture).

[0668]

[0669] Table 9. Relative amino acid profiles of the water-wampee protein isolate produced on a pilot-scale basis.

[0670]

[0671]

[0672] * indicates an essential amino acid.

[0673] Example 2C

[0674] Water-wheat peel protein isolate was prepared by membrane filtration.

[0675] This embodiment demonstrates the extraction of protein from defatted and debittered water pomegranate pulp via membrane filtration and the production of a protein-rich water pomegranate composition (water pomegranate protein isolate), and generally follows... Figure 2C The exemplary method described herein.

[0676] In this embodiment, defatted and debittered water yellow peel meal was used as the starting material for protein extraction. Except for ethyl acetate as the solvent, the defatted and debittered water yellow peel meal was obtained according to the method described in Example 1 above.

[0677] Aqueous slurry of defatted and debittered water-derived yellow peel pulp was prepared using water (1:6; 15% solids) in a high-shear mixer. The pH of the slurry was adjusted to pH 8 with 2M NaOH (~8% aqueous solution) and continuously stirred at 25°C for 2 hours. The slurry was separated into a protein-containing liquid phase and a wet filter cake using a decanter. A 10kDa molecular weight cutoff (MWCO) hollow fiber membrane module (420cm³) with a laboratory-scale membrane filtration device was used. 2 ) or 5kDa MWCO flat panel box (1000cm) 2 Filter the liquid phase containing proteins. Select the permeate flow rate and transmembrane pressure (~2.8 bar) to obtain a reasonable permeate flux. Perform membrane filtration at a concentration factor (CF) of 4–5 and a permeate factor (DF) of 2–4. The obtained retentate is further washed and freeze-dried into a protein isolate powder.

[0678] This method results in the extraction of approximately 70-75% by weight of the total protein from the meal and the recovery of approximately 30% by weight. The aqueous yellow peel protein isolate produced by membrane filtration contains approximately 80% by weight of protein.

[0679] In one experiment, tests involving 5kDa or 10kDa membranes were conducted on a protein-containing liquid phase (generated according to the protocol described above). Conditions and results are provided in Table 10 below.

[0680] Table 10. Summary of protein purity and yield after membrane filtration

[0681]

[0682] Example 3

[0683] Molecular weight characteristics of water huangpi protein

[0684] This embodiment illustrates the molecular weight characteristics of proteins present in (i) water-yellow peel soybeans, (ii) cold-pressed water-yellow peel meal, (iii) defatted and debittered water-yellow peel meal obtained according to the method described in Example 1 above, and (iv) a protein-rich water-yellow peel composition obtained according to the method described in Examples 2A-2C above. The molecular weights of water-yellow peel proteins obtained after different processing stages are shown compared to the molecular weight characteristics of soybean protein isolates and proteins extracted from partially defatted soybean meal.

[0685] refer to Figures 5A-5D The size distribution and relative abundance of proteins in water-yellow-skinned beans and derived materials were determined by SDS-PAGE. Molecular weights were typically determined using the following protocol. Figure 5A Protein extracts in these groups were prepared by mechanical disruption of specified materials in a protein extraction buffer containing 50 mM TRIS-HCl (pH 8.3), 100 mM NaCl, 2 mM EDTA, 1% SDS, and 1 mM PMSF. Protein concentration in each extract was determined using the Bradford assay with bovine serum albumin (BSA) as the standard. Extracts were diluted and mixed with denaturing SDS-PAGE sample buffer before loading onto 12% SDS-PAGE gels (approximately 30 μg per lane). Pure BSA from commercial stock was diluted directly into the SDS-PAGE sample buffer and included as an unstained molecular weight marker (approximately 66 kDa) and a protein content reference (approximately 6 μg per lane). Soy protein isolates were soybean protein products isolated from SUPRO XT40 from Solae (10002061). Reduced-fat soybean meal was prepared in-house using cold-pressed commercially available soybeans (soya). The prestained molecular weight standard (not shown) was ThermoScientific PageRuler Plus Prestained Protein Ladder (26619). Reference Figures 5B-5DProtein samples in these groups were taken from various stages of the preparation of compositions rich in Aquamarine protein. Lyophilized (FD) powders for Aquamarine protein concentrates, separated by isoelectric precipitation or membrane filtration, were reconstituted in water at 20 mg / ml (2% w / v). Protein concentrations in each sample were determined by Bradford or BCA assays using bovine serum albumin (BSA) as a standard. Aliquots were diluted in H2O and mixed with denaturing SDS-PAGE sample buffer before loading the protein samples onto SDS-PAGE gels. Note that for groups comparing the protein profiles in lyophilized isolates obtained by membrane filtration of a simple pH 8 extract with those of the corresponding “parental” lyophilized pH 8 extract (concentrate), relatively fewer proteins were loaded per lane, and different gel systems were used.

[0686] Several easily identifiable proteins were found in water hyacinth beans, ranging in size from about 10 kDa to 250 kDa. The single most abundant protein type (representing 30-40% of the total protein) was bimodal at about 55 kDa. In addition, five other significant categories were observed at 250 kDa, 130 kDa, 25 kDa, 15 kDa, and 10 kDa. These six categories, likely corresponding to the protein storage proteins in water hyacinth beans, have been found to have the greatest impact on the functionality of water hyacinth bean meal and flour, as well as the protein concentrates or isolates prepared from them. Throughout the processing steps for producing raw flour as described in the preceding examples, the major proteins (and most other proteins) found in water hyacinth beans were observed to remain largely intact. Importantly, these proteins were observed to be readily extracted from defatted and / or debittered water hyacinth bean meal using the aqueous extraction, isoelectric precipitation, or membrane filtration methods described herein (Examples 2A-2C).

[0687] Example 4

[0688] Functional properties of water huangpi protein

[0689] In this embodiment, the solubility, viscosity, and emulsification properties of the water yellow peel protein composition produced according to the method described in Example 2 above were characterized and compared with soybean, pea, lupin, and sunflower seed proteins.

[0690] Protein solubility

[0691] To measure the solubility of the water citrus protein composition, a 2% w / w protein solution (based on nitrogen*5.7) was prepared in water, adjusted to the specified pH using acid or base, and stirred at room temperature for 2 hours. The sample was centrifuged at 20,000 g for 10 min at 20°C, and the supernatant was collected. The nitrogen content of the supernatant was determined using the Kjeldahl method. Protein solubility (e.g., protein present in 20,000 g of supernatant) is expressed as a percentage of the initial amount of protein added to the solution. Protein solubility can also be expressed as the mass of solute dissolved per volume of solvent (g / L).

[0692] Figure 3 Protein solubility profiles of *Aquamarine* protein concentrates in water at various pH values ​​are shown. Solubility profiles were prepared by adjusting the pH of a 2% w / w protein aqueous solution (based on nitrogen* 5.7) to the desired value (pH 3 to pH 9) with HCl or NaOH. The suspensions were agitated for 2 hours at room temperature and then centrifuged to remove insoluble material. Figure 4A and 4D The solubility of proteins in concentrated or isolated *Phellodendron amurense* protein extracts was compared with that of proteins in commercial plant protein compositions at pH 7.0.

[0693] Viscosity

[0694] To measure the viscosity of the protein-rich water-based yellow peel composition, a 4% w / w protein solution (based on nitrogen*5.7) was prepared and stirred at room temperature for 30 minutes. The protein solution was then heated at 90°C for 15 minutes and cooled to room temperature. Viscosity was measured using a rheometer at 20°C in 0s. -1 up to 1000s -1 Shear rate measurement. Figure 4B and 4E A comparison was made between solutions prepared from concentrated or isolated *Phellodendron amurense* protein extract and solutions prepared from commercial plant protein compositions at 100 s. -1 Viscosity at the shear rate.

[0695] Protein emulsification

[0696] The emulsion was prepared using a protein-to-fat ratio of 1:10. A 1% protein concentration and a 10% sunflower oil aqueous solution were used. First, the protein was hydrated, and then the fat was added slowly while mixing at high shear (15,000 rpm) for 2 minutes. The water-protein-oil mixture was homogenized at 300 / 30 bar to form a stable emulsion. The droplet size of the emulsion was analyzed by laser diffraction. The observed droplet sizes are shown in... Figure 4C and 4E middle.

[0697] The functional properties results of solubility, viscosity, and emulsifying properties are shown in Figures 4A-4F In comparison with commercially available legume proteins tested, the aquaponica protein concentrate or isolate was found to have superior solubility (approximately 80%). The viscosity and emulsifying properties of the aquaponica protein (concentrate or isolate) were found to be comparable to those of pea and soybean proteins.

[0698] Example 5

[0699] Further functional studies of proteins in water huangpi

[0700] This embodiment details the functional evaluation of the aquatic yellow peel protein isolate prepared generally according to the protocols provided in Examples 2B, running A and B (sample B1). The emulsifying properties, viscosity, water-holding capacity, oil-holding capacity, gelling properties, foaming properties, powder dispersibility, and solubility at pH 7 of the resulting aquatic yellow peel protein isolate were evaluated compared to similar properties observed in commercially available soybean and pea protein isolates.

[0701] Protein solubility

[0702] Solubility was determined at pH 7 for protein suspensions with a 2% protein content, and the solubility of the supernatant after centrifugation at 15000g for 10 minutes was evaluated using the Kjedahl method.

[0703] Table 11 shows the observed solubility of the aquatic radix protein isolates. The two aquatic radix protein isolates exhibited high protein solubility of 38% and 57%, respectively. The observed solubility was significantly higher than that of pea protein and comparable to (or higher than) that of soybean protein in run B1 aquatic radix protein isolate.

[0704] Viscosity

[0705] To measure the viscosity of the protein-rich water-based yellow peel composition, a 10% w / w protein solution (based on nitrogen*5.7) was prepared and stirred. A rheometer was used at 25°C with a stirring speed of 0.1 s⁻¹. -1 up to 1000s -1 Viscosity was measured at a certain shear rate.

[0706] Table 11 shows the observed viscosity of the *Phellodendron amurense* protein isolate. The *Phellodendron amurense* protein isolate exhibited a viscosity with a relatively constant shear rate, corresponding to Newtonian behavior. The viscosity of the *Phellodendron amurense* protein isolate was observed to be relatively low, approximately 10. -2 Pa·s, which is slightly higher than the viscosity of water and comparable to that of pea protein isolate.

[0707] Figure 6AThe viscosity of the aqueous wampee protein isolate solution (obtained from run B1) measured at different shear rates is shown and compared with solutions of prepared pea protein isolate or soybean protein isolate. At all measured shear rates, the aqueous wampee protein isolate exhibited a lower viscosity than both the pea protein isolate and the soybean protein isolate.

[0708] Table 11. Protein solubility of water hyacinth protein, pea protein and soybean protein isolates

[0709]

[0710] Protein emulsification

[0711] The emulsifying properties of protein samples were measured by generating an oil-in-water emulsion. A solution containing 1% protein was prepared in water. The emulsion was generated by mixing the protein solution with oil at a 75 / 25 ratio followed by sonication. The oil droplet size distribution was then measured on a Mastersizer (Malvern) particle size analyzer using two dispersants (water and SDS) according to procedure PR-14010. The refractive index of sunflower oil was 1.46, and that of water was 1.33. The absorbance index of sunflower oil was 0.01.

[0712] For emulsification assessment, the emulsification characteristics of the water wampee protein isolate, pea protein isolate, and soybean protein isolate were evaluated immediately after preparation (day 0) and immediately after 7 days of storage (day 7). Table 12 shows the particle size distribution D50 values ​​of the water wampee protein isolate observed on day 0 and day 7. As provided herein, the D50 value represents the droplet size at which 50% of the particles in the sample are larger than a specified value.

[0713] The water citrus protein isolate was observed to form a fine emulsion (median size less than 5 μm) immediately after preparation and remained stable after 7 days of storage. The water citrus protein sample exhibited very good emulsifying properties, similar to those of soybean protein and casein. Figure 6B The droplet size distribution of the water yellow peel protein isolate emulsion (prepared on a pilot scale) is shown compared to emulsions prepared using sodium caseinate (as a reference), pea protein isolate, or soybean protein isolate. Figure 6B As shown, the droplet size distribution of the water-yellow peel emulsion is unimodal and similar to the median droplet size and droplet size distribution of sodium caseinate.

[0714] Table 12. Emulsion stability and D50 value

[0715]

[0716]

[0717] Water holding capacity and oil holding capacity

[0718] The water-holding and oil-holding capacities of the aquamarine protein isolate were measured by adding each sample to oil or water at a concentration of 20 mg / ml dry matter. The suspension was mixed with agitation for 1 hour. After centrifugation at 15000g for 10 minutes, the water and oil content in the precipitate was measured and compared with the initial weight of the material. The results are expressed as the number of times the sample was able to retain its weight in water or oil. As shown in Table 13, the aquamarine protein isolate exhibited moderate water-holding capacity, but lower than that of pea and soybean protein isolates; compared with soybean and pea proteins, the aquamarine protein isolate had slightly higher oil-binding properties.

[0719] Table 13. Water-holding capacity and oil-holding capacity

[0720]

[0721] Foaming properties

[0722] Foaming properties were assessed using Foamscan (Teclis Scientific) with a 0.1% w / v protein solution (60 mL) at pH 7. Foam was formed by aerating air into the solution at a flow rate of 200 mL / min for 30 seconds. Foam volume and its stability were then recorded over a 10-min period. Egg white was used as a reference for this test. Table 14 shows the results of egg white (as a reference) producing a large volume of foam, which was very stable over time. Water-based protein produced a large volume of foam, but the foam volume decreased significantly over time.

[0723] Table 14. Foaming Properties

[0724]

[0725] gelling properties

[0726] The minimum gel concentration was measured by preparing solutions with a protein content of 2% to 20% in test tubes. After solubilization, the solutions were heated in a water bath at 85°C for 1 hour and then cooled at 4°C for 2 hours. If the protein solution behaved like a liquid (i.e., free-flowing) before heating but did not flow when the test tube was inverted after heating, the protein solution was considered to have formed a gel.

[0727] Table 15 shows the minimum gelation concentration results for the water citrus peel protein isolate. The water citrus peel protein isolate was found to exhibit gelation comparable to that of pea and soybean proteins.

[0728] Powder dispersibility

[0729] The powder dispersibility was measured as follows. Five (5) g of sample were added to 100 ml of water under 500 rpm (vortex) mixing. The dispersion was mixed for 5 min and then filtered through a 30 μm pore size filter. The filter and any retained contents were dried at 105 °C for 4 h and weighed. The proportion of material (undispersed product) retained on the filter per g of sample was calculated.

[0730] Table 15 shows the dispersibility results of the water yellow peel protein isolate. As shown in Table 15, water yellow peel protein was observed to have excellent dispersibility compared to soybean and pea proteins.

[0731] Table 15. Dispersibility and gelling properties

[0732]

[0733] "--" indicates Example 6 that was not measured.

[0734] Various food uses and levels of protein-rich water chestnut extract

[0735] The protein-rich water chestnut compositions described herein (e.g., water chestnut protein concentrates or isolates) can be used as direct protein alternatives to animal or plant proteins in a wide variety of conventional food and beverage products across multiple categories. Exemplary food categories and levels of use are summarized in Table 16 below.

[0736] Table 16. Exemplary Foods and Usage Levels

[0737]

[0738]

[0739] Example 7

[0740] Plant-based milk beverages

[0741] This embodiment describes the preparation of a plant-based milk beverage using the protein-rich water citrus composition (e.g., water citrus protein concentrate or isolate) described herein, including the protein composition obtained according to the methods in Examples 2A-2C above.

[0742] A protein milk is prepared by hydrating a protein-rich *Amanita muscaria* (a type of wild fruit) composition, for example, 5.5-6% by weight of *Amanita muscaria* protein isolate (70-80 wt% protein) (per serving of 10 g protein), in hot water (e.g., 140-160°F) for approximately 15-20 minutes using a high-shear mixer. Rapeseed oil / soybean oil, sugar, thickener, and flavoring are added to the hydrated aqueous protein and mixed for another 5-10 minutes. The mixture is then homogenized to form a uniform emulsion and pasteurized.

[0743] Example 8

[0744] Plant-based yogurt

[0745] This embodiment describes the preparation of plant-based yogurt using a protein-rich water citrus composition (e.g., water citrus protein concentrate or isolate) as described herein, including protein compositions obtained according to the methods in Examples 2A-2C above.

[0746] Aquamarine protein milk is prepared by hydrating a protein-rich aquamarine composition, such as 9-10% by weight aquamarine protein isolate (70 wt% protein), in hot water (e.g., 140-160°F) for approximately 15-20 minutes using a high-shear mixer. Other optional ingredients, such as rapeseed / soybean oil, sugar, thickeners, and flavorings, are added to the hydrated aqueous protein and mixed for another 5-10 minutes. The mixture is then homogenized into a uniform emulsion, pasteurized, and cooled to approximately 100°F. A vegan yogurt culture is added and fermented for approximately 6-10 hours. Once the yogurt reaches the desired pH of 4.5, it is stirred, poured into containers, and refrigerated.

[0747] Example 9

[0748] Fortified white bread

[0749] This embodiment describes the production of white bread fortified with a protein-rich water citrus composition (e.g., water citrus protein concentrate or isolate) as described herein, including the protein composition obtained according to the methods in Examples 2A-2C above.

[0750] Water-fortified wampee protein concentrate: Two bread doughs were prepared: (1) a control dough without protein fortification (3 g protein per serving); and (2) a test dough with a protein-rich water-fortified wampee composition (6 g protein per serving). In the test dough, at least a portion (20 wt%) of the wheat flour was replaced with a protein-rich water-fortified wampee composition (e.g., concentrate). Other optional ingredients in the formulation included salt, sugar, yeast, oil, butter, skim milk, and water. The doughs were mixed, weighed, shaped, placed in a pan, proofed, and baked at approximately 420°F for approximately 25–30 minutes. The breads were then evaluated. The volume, texture, and flavor of the control and fortified breads were evaluated.

[0751] Aquamarine Protein Isolate: Two bread doughs were prepared: (a) Control – no added protein (b) Protein-Rich – containing aquamarine protein. In the protein-rich recipe, wheat flour was replaced with 10 wt% of a protein-rich aquamarine composition (aquamarine protein isolate, approximately 70-80 wt% protein). Optional ingredients included sugar, salt, butter, yeast, and water. The ingredients were mixed into a dough, weighed, shaped, and placed on a baking sheet, proofed, and baked at approximately 420℉ for approximately 25-30 minutes.

[0752] Example 10

[0753] Fortified biscuits

[0754] This embodiment describes the production of biscuits fortified with a protein-rich water citrus composition (e.g., water citrus protein concentrate or isolate) as described herein, including the protein composition obtained according to the methods in Examples 2A-2C above.

[0755] Two types of biscuit dough were prepared: (1) a control dough without protein fortification (3 g protein per serving); and (2) a test dough with a protein-rich aquamarine composition (5 g protein per serving). In the test dough, a portion (20%) of the wheat flour was replaced with a protein-rich aquamarine composition (e.g., aquamarine protein concentrate, approximately 50 wt% protein). Other optional ingredients in the formulation included salt, sugar, sesame seeds, oil, leavening agents (e.g., sodium bicarbonate), and water. The dough was prepared, pressed to the desired thickness (~1.5 mm), cut into the desired shapes, and baked. The texture and flavor of the control and fortified biscuits were evaluated.

[0756] Wheat biscuits with added citrus protein or soy protein

[0757] Three types of biscuit dough were prepared: (A) a control dough without protein fortification (2.1 g protein per serving); (B) a test dough with a protein-rich aquamarine composition (4.9 g protein per serving, including 2.9 g protein from aquamarine protein isolate); and (C) a test dough rich in soy protein (2.7 g protein from soy protein). The control dough was prepared using a 50:50 blend of whole wheat and all-purpose flour. In the test dough (B) using aquamarine protein isolate, the whole wheat / all-purpose flour blend was replaced by 8.7% aquamarine protein isolate; in the test dough (C) using a soy protein enrichment, the whole wheat / all-purpose flour blend was replaced by 7.4% soy protein isolate. Other ingredients in the formulation included salt, sugar, malt barley, canola oil, corn starch, baking soda, and water. Additional water was added to both test doughs to achieve a texture similar to the control dough; all other ingredients except flour remained unchanged in both the control and test doughs. Prepare the dough, roll it to the desired thickness (~1.5 mm), cut it into 1.5-inch squares, and bake. Evaluate the texture and taste of the control and fortified cookies. Table 17 below shows the sensory evaluation summary.

[0758] Fortified biscuits containing hyaluronic acid have a deeper color and a richer whole-wheat flavor than the control biscuits.

[0759] Table 17. Sensory Evaluation

[0760]

[0761]

[0762] Example 11

[0763] Plant-based ready-to-drink protein beverage

[0764] This embodiment describes the preparation of a plant-based ready-to-drink (RTD) chocolate protein beverage using a protein-rich water citrus composition (e.g., water citrus protein isolate) as described herein, including the protein composition obtained according to the methods in Examples 2A-2C above.

[0765] Three ready-to-drink chocolate beverages were prepared: (A) with 16 g / serving of aquamarine protein, (B) with 20 g / serving of aquamarine protein, and (C) with 20 g / serving of pea protein. First, cocoa was hydrated separately in hot water. Aquamarine protein was hydrated with dipotassium phosphate in warm water for 15 minutes. Hydrated cocoa, sunflower oil, lecithin, sugar, natural sweetener, thickener, and flavoring agent were added to the hydrated aquamarine protein and mixed for another 5 minutes. The mixture was then homogenized to form a uniform emulsion, pasteurized, bottled, and stored at refrigerated temperatures for further evaluation.

[0766] The following characteristics of the beverages were evaluated: visual appearance (including color, physical appearance, and stability), aroma, texture and mouthfeel (including creamy, smooth, gritty, chalky, thick, and thin), flavor and taste (including sweetness, saltiness, aftertaste, and off-flavors), and overall preference (acceptable / unacceptable). Table 18 below shows a summary of the sensory evaluation. Both water-based yellow-skin protein beverages were preferred over the pea protein beverage.

[0767] Table 18. Sensory Assessment Summary – Ready-to-Drink Chocolate Protein Beverage

[0768]

[0769] Example 12

[0770] Protein powder beverage mixture

[0771] This embodiment describes the preparation of a powdered chocolate protein beverage mixture using a protein-rich water citrus composition (e.g., water citrus protein isolate) as described herein, including a protein composition obtained according to the methods in Examples 2A-2C above.

[0772] This embodiment describes the preparation of a powdered chocolate protein beverage mixture using a protein-rich water citrus composition (e.g., water citrus protein isolate) as described herein, including the protein composition obtained according to the methods in Examples 2A-2C above.

[0773] Two chocolate protein powder mixtures were prepared – (1) 15g of aquamarine protein and (2) 15g of pea protein, per serving. Other ingredients included cocoa, sugar, natural sweeteners, salt, and flavorings. All ingredients were added to a blender and mixed for 10 minutes until all ingredients were homogeneous. The products were packaged in metallized bags for further use. The powdered products were mixed with 12 fl oz of water and used for sensory evaluation. The following characteristics of the reconstituted aquamarine and pea protein beverages were evaluated: visual appearance (including color, physical appearance, and stability), aroma, texture and mouthfeel (including creamy, smooth, gritty, chalky, thick, and thin), flavor and taste (including sweet, salty, aftertaste, and off-taste), and overall preference (acceptable / unacceptable). A summary of the sensory evaluation is shown in Table 19 below. The aquamarine chocolate powder was preferred over the chocolate pea protein powder.

[0774] Table 19. Sensory Evaluation Summary - Chocolate Protein Beverage Mixture (Reconstructed)

[0775]

[0776] As used herein, the term "about" refers to the typical range of error for various values ​​that is readily known to those skilled in the art. The reference herein to "about" a value or parameter includes (and describes) an implementation for that value or parameter itself. For example, "about x" includes and describes "x" itself. In some implementations, the term "about," when used in conjunction with a measurement or to modify a value, unit, constant, or range of values, refers to a variation of + / - 5% of said value or parameter.

[0777] The phrase "between two values ​​or parameters" as used herein includes (and describes) implementations that include the two values ​​or parameters themselves. For example, a description of "between x and y" includes a description of "x" and "y" themselves.

Claims

1. A method of producing a protein-enriched pongamia ingredient comprising: combining fat-reduced pongamia meal with an alkyl alkanoate solvent to produce a pongamia meal, wherein the alkyl alkanoate solvent comprises ethyl acetate, and wherein the pongamia meal is defatted and debittered and has (i) less than 200 ppm of pongamol; or (ii) less than 200 ppm of pongamidione; preparing an aqueous slurry of the pongamia meal; adjusting the pH of the aqueous slurry to a pH between 8 and 10; separating the slurry into a protein liquid fraction and an insoluble wet cake fraction; neutralizing, concentrating, and / or pasteurizing the protein liquid fraction; and drying the protein liquid fraction to provide a protein-enriched pongamia ingredient, wherein the pongamia ingredient comprises: at least 45% pongamia protein on a dry weight basis; less than 50% carbohydrate on a dry weight basis; less than 5% fat on a dry weight basis; less than 200 ppm pongamol; and less than 200 ppm pongamidione.

2. The method of claim 1, wherein the pongamia meal has less than 200 ppm of pongamol and pongamidione combined.

3. The method of claim 1, wherein the protein-enriched pongamia ingredient comprises at least 50% pongamia protein on a dry weight basis.

4. A method of producing a protein-enriched pongamia ingredient comprising: combining fat-reduced pongamia meal with an alkyl alkanoate solvent to produce a pongamia meal, wherein the alkyl alkanoate solvent comprises ethyl acetate, and wherein the pongamia meal is defatted and debittered and has (i) less than 200 ppm of pongamol; or (ii) less than 200 ppm of pongamidione; preparing an aqueous slurry of the pongamia meal; adjusting the pH of the aqueous slurry to a pH between 8 and 10, separating the slurry into a protein liquid fraction and an insoluble wet cake fraction; precipitating at least a portion of the pongamia protein from the protein liquid fraction to obtain purified pongamia protein solids; neutralizing and pasteurizing the purified pongamia protein solids; and drying the purified pongamia protein solids to provide a protein-enriched pongamia ingredient, wherein the pongamia ingredient comprises: at least 45% pongamia protein on a dry weight basis; less than 50% carbohydrate on a dry weight basis; less than 5% fat on a dry weight basis; less than 200 ppm pongamol; and less than 200 ppm pongamidione.

5. The method of claim 4, wherein the pongamia meal has less than 200 ppm of pongamol and pongamidione combined.

6. The method of claim 4, further comprising washing the pongamia protein solids prior to drying.

7. The method of any one of claims 4-6, wherein the precipitating step is performed by isoelectric precipitation.

8. The method of any one of claims 4 to 6, wherein the protein-enriched pongamia ingredient comprises at least 70% pongamia protein on a dry weight basis.

9. The method of any one of claims 4 to 6, further comprising: dehulling the pongamia beans to produce dehulled pongamia beans; and pressing the dehulled P. juliflora legumes to remove at least a portion of free oil in the P. juliflora legumes to produce a reduced fat P. juliflora meal.

10. The method of claim 9, further comprising grinding the reduced fat P. juliflora meal prior to combining with the solvent.

11. A method of producing a protein-enriched P. juliflora ingredient, comprising: combining the reduced fat P. juliflora meal with an alkyl ester of a chain alkanoic acid solvent to produce a P. juliflora meal, wherein the alkyl ester of a chain alkanoic acid solvent comprises ethyl acetate, and wherein the P. juliflora meal is defatted and debittered and has (i) less than 200 ppm picral; or (ii) less than 200 ppm phyllatone; preparing an aqueous slurry of the P. juliflora meal; adjusting the pH of the aqueous slurry to a pH between 8 and 10, separating the slurry into a protein liquid fraction and an insoluble wet cake fraction; passing the protein liquid fraction through a membrane system to obtain a retentate comprising P. juliflora protein; optionally washing, neutralizing, and / or pasteurizing the retentate; and drying the retentate to provide a protein-enriched P. juliflora ingredient, wherein the P. juliflora ingredient comprises: at least 45% P. juliflora protein on a dry weight basis; less than 50% carbohydrate on a dry weight basis; less than 5% fat on a dry weight basis; less than 200 ppm picral; and less than 200 ppm phyllatone.

12. The method of claim 11, wherein the protein-enriched P. juliflora ingredient comprises at least 70% P. juliflora protein on a dry weight basis.

13. The method of claim 11 or claim 12, further comprising: dehulling P. juliflora legumes to produce dehulled P. juliflora legumes; and pressing the dehulled P. juliflora legumes to remove at least a portion of free oil in the P. juliflora legumes to produce a reduced fat P. juliflora meal.

14. The method of claim 13, further comprising grinding the reduced fat P. juliflora meal prior to combining with the solvent.

15. A method of producing a protein-enriched P. juliflora ingredient, comprising: combining the reduced fat P. juliflora meal with an alkyl ester of a chain alkanoic acid solvent to produce a P. juliflora meal, wherein the alkyl ester of a chain alkanoic acid solvent comprises ethyl acetate, and wherein the P. juliflora meal is defatted and debittered and has (i) less than 200 ppm picral; or (ii) less than 200 ppm phyllatone; preparing an aqueous slurry of the P. juliflora meal; adjusting the pH of the aqueous slurry to a pH between 4 and 5 to obtain P. juliflora protein solids; washing, neutralizing, and pasteurizing the purified protein solids; and drying the purified P. juliflora protein solids to provide a protein-enriched P. juliflora ingredient, wherein the P. juliflora ingredient comprises: at least 45% P. juliflora protein on a dry weight basis; less than 50% carbohydrate on a dry weight basis; less than 5% fat on a dry weight basis; less than 200 ppm picral; and less than 200 ppm phyllatone.

16. A protein-enriched P. juliflora ingredient produced by the method of any one of claims 1-15, comprising: at least 45% P. juliflora protein on a dry weight basis; less than 50% carbohydrate on a dry weight basis; less than 5% fat on a dry weight basis; less than 5% fat on a dry weight basis; less than 200 ppm picraline; and less than 200 ppm picralidine.

17. The ingredient of claim 16, wherein at least 35% of the protein present in the ingredient is soluble in water at a pH of at least pH 6.

18. The ingredient of claim 16, wherein the ingredient has less than 100 ppm combined picraline and picralidine.

19. The ingredient of claim 16, wherein the ingredient has between 45% and 70% picraline protein on a dry weight basis.

20. The ingredient of claim 16, wherein the ingredient is a picraline protein concentrate.

21. The ingredient of claim 16, comprising at least 70% picraline protein on a dry weight basis.

22. The ingredient of claim 21, wherein the ingredient has less than 200 ppm combined picraline and picralidine.

23. The ingredient of claim 21, wherein the ingredient has between 70% and 90% picraline protein on a dry weight basis.

24. The ingredient of claim 23, wherein the ingredient is a picraline protein isolate.

25. The ingredient of any one of claims 16 to 24, wherein the ingredient is derived from picraline meal, wherein the protein-enriched picraline ingredient has at least 1.25 times greater picraline protein content than the picraline meal.

26. The ingredient of any one of claims 16 to 24, wherein the ingredient has less than 4% fat on a dry weight basis.

27. The ingredient of any one of claims 16 to 24, wherein the ingredient has less than 2% fat on a dry weight basis.

28. The ingredient of any one of claims 16 to 24, wherein the ingredient has a relative amino acid profile comprising at least 15% glutamic acid, at least 12% aspartic acid, at least 9% leucine, at least 8% lysine, at least 6% phenylalanine, or any combination thereof.

29. The ingredient according to any one of claims 16 to 24, wherein the ingredient has a viscosity of at least 2 mPa*s at a shear rate of 100 s -1 .

30. The ingredient of any one of claims 16 to 24, wherein the ingredient, when emulsified, produces an emulsion having an average droplet size of at least 1 μιη.

31. The ingredient of any one of claims 16 to 24, wherein the ingredient has a corrected amino acid score for protein digestibility of at least 0.

7.

32. The ingredient of any one of claims 16 to 24, wherein the ingredient has an average molecular weight of protein between 10,000 and 250,000 Daltons.

33. The ingredient of any one of claims 16 to 24, wherein the ingredient comprises seed storage proteins, and wherein 30-40% of the protein present is protein having a molecular weight between 45 kDa and 70 kDa as determined by SDS-PAGE.

34. The ingredient of claim 33, wherein the ingredient further comprises a seed storage protein having a molecular weight of 170-250 kDa, 115-160 kDa, 45-70 kDa, 19-25 kDa, 14-17 kDa, or 10-13 kDa, or any combination thereof.

35. The ingredient of any one of claims 16-24, wherein the ingredient has: (i) a viscosity between 2 mPa*s and 100 mPa*s at a shear rate of 100 s -1 - 1 mPa*s. (ii) a foaming capacity of between 100% and 200% of the volume of the 0.1% protein solution; (iii) a bulk density of at least 0.2 g / cm3 3 of at least 0.2 g / cm3 (iv) a protein solubility of at least 35% at pH 7; (v) a median emulsion droplet size of less than or equal to 5 pm; (vi) a median emulsion droplet size of less than or equal to 5 pm after 7 days of storage; (vii) a water holding capacity of at least 1.5 g water per g of the protein- enriched G. biloba ingredient; (viii) an oil holding capacity of at least 1.5 g oil per g of the protein- enriched G. biloba ingredient; (ix) a minimum gelling concentration of at least 10 g of the protein- enriched G. biloba ingredient per 100 g; (x) a powder dispersibility of at least 10%; or (xi) a neutral, non-bitter taste; or any combination of (i)-(x) thereof.

36. The ingredient of claim 35, wherein the ingredient has: (i) a viscosity between 2 mPa*s and 100 mPa*s at a shear rate of 100 s -1 - 1 mPa*s. (ii) a foaming capacity of between 100% and 200% of the volume of the 0.1% w / v G. biloba protein solution; (iii) a bulk density of at least 0.2 g / cm3 3 of at least 0.2 g / cm3 (iv) a protein solubility of at least 35% at pH 7; (v) a median emulsion droplet size of less than or equal to 5 pm; (vi) a median emulsion droplet size of less than or equal to 5 pm after 7 days of storage; (vii) a neutral, non-bitter taste; or any combination of (i)-(vii) thereof.

37. The ingredient of claim 35, wherein the ingredient has: (i) a viscosity between 2 mPa*s and 100 mPa*s at a shear rate of 100 s -1 - 1 mPa*s. (ii) a foaming capacity of between 100% and 200% of the volume of the 0.1% w / v protein solution; (iii) a bulk density of at least 0.2 g / cm3 3 of at least 0.2 g / cm3 (iv) a protein solubility of at least 35% at pH 7; (v) a median emulsion droplet size of less than or equal to 5 pm; (vi) a median emulsion droplet size of less than or equal to 5 pm after 7 days of storage; (vii) a water holding capacity of at least 1.5 g water per g of the protein- enriched G. biloba ingredient; (viii) a minimum gelling concentration of at least 10 g of the protein- enriched G. biloba ingredient per 100 g; or (ix) a neutral, non-bitter taste or any combination of (i)-(ix) thereof. (i) a bulk density of at least 0.2 g / cm3 3 of at least 0.2 g / cm3 38. The ingredient of claim 35, wherein the ingredient has: (ii) a protein solubility of at least 35% at pH 7; (iii) a water holding capacity of at least 1.5 g water per g of the protein- enriched G. biloba ingredient; (iv) an oil holding capacity of at least 1.5 g oil per g of the protein- enriched G. biloba ingredient; (v) a minimum gelling concentration of at least 10 g of the protein- enriched G. biloba ingredient per 100 g; or (vi) a neutral, non-bitter taste or any combination of (i)-(vi) thereof.

39. The ingredient of claim 35, wherein the ingredient has: (i) a foaming capacity of between 100% and 200% for 0.1% w / v protein solution volume; (ii) a minimum gel concentration of at least 7 g of protein-rich pongamia ingredient per 100 grams; or (iii) neutral, no bitter taste; or any combination of (i)-(iii) thereof.

40. A food product, beverage product, dietary supplement product, or other product, comprising: The protein-rich pongamia ingredient of any one of claims 16 to 39.

41. The product of claim 40, wherein the product is a baked good, a protein supplement, a protein bar, or a non-dairy beverage.

42. The product of claim 40, wherein the product is a medical food, an infant formula, a cosmetic, or a pharmaceutical product.

43. The product of claim 40, wherein the product is a beverage product, a dairy alternative, a meat alternative product, or an egg alternative.

44. The product of claim 43, wherein the beverage product is a fruit smoothie, a meal replacement beverage, a protein beverage, a instant milkshake.

45. The product of any one of claims 40, 43, and 44, wherein the product is a beverage product comprising at least 20 g of protein-rich pongamia ingredient per serving.

46. The product of claim 43, wherein the dairy alternative is a non-dairy milk, a non-dairy cheese, a non-dairy coffee whitener or creamer, a non-dairy yogurt, a non-dairy Greek yogurt, a non-dairy drinkable yogurt.

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