Methods of extracting high-quality protein concentrates, isolates and fiber fractions, compositions, and their applications in food products

Abstract

The present invention relates to extracting and processing protein concentrates, isolates, and a protein-fiber fraction from cluster bean, also known as guar beans (botanical name Cyamopsis tetragonoloba). The disclosed hexane-free, aqueous-ethanol extraction process, combined with controlled low-temperature (<75 °C) vacuum desolventization to preserve light color, bland flavor, and protein functionality, provides a sustainable, clean-label method for producing cluster bean protein concentrates and isolates suitable for direct human foods. This approach avoids the high heat and reverse-polarity extraction seen in prior art and enables food-grade, industrially scalable CPC and CPI.

Description

[0001] VHPM 00773. OOlWOl METHODS OF EXTRACTING HIGH-QUALITY PROTEIN CONCENTRATES, ISOLATES AND FIBER FRACTIONS, COMPOSITIONS PRODUCED THEREFROM,

[0002] AND THEIR APPLICATIONS IN FOOD PRODUCTS

[0003] CROSS-REFERENCE TO RELATED APPLICATION

[0004] This application claims priority to United States Provisional Application Number 63 / 724,110 that was filed on November 22, 2024. The entire content of the application referenced above is hereby incorporated by reference herein.

[0005] BACKGROUND

[0006] Cluster beans (Cyamopsis tetragonolobdy also known as Guar, primarily cultivated in India and Pakistan, are a high-value legume recognized for their use in producing guar gum, which finds widespread application across various industries, including food, cosmetics, pharmaceuticals, oil fracking, and other industrial sectors. The cluster bean plant is valued for its drought resistance and nitrogen-fixing capability, making it an important crop for sustainable agriculture, particularly in arid regions. The processing of cluster bean seeds to produce guar gum yields valuable byproducts, notably guar korma and churi, which are rich in protein and fiber. These byproducts hold considerable untapped potential for upscaling to human nutrition.

[0007] In traditional Indian cuisine, cluster beans are consumed in various forms, including ki dal (split bean soup), ki sabzi (stir-fried beans), and ke pakode (fried bean fritters). Despite these culinary uses, extracting high-purity protein concentrates and isolates from cluster bean has faced substantial challenges. Key commercialization obstacles include anti -nutritional factors (ANFs), undesirable odors, bitter taste, limited scalability, and difficulty in effectively removing impurities. Consequently, there is currently no large-scale production of Cluster Bean protein concentrates or isolates that meet the quality standards required for human consumption.

[0008] The Cluster Bean Germ or Guar korma, a nutrient-dense byproduct from guar gum processing, contains significant quantities of protein and fiber. Traditionally, guar korma has been mainly toasted and incorporated into animal feed formulations due to its approximately 50-58% protein content. The toasting process eliminates ANFs, such as trypsin inhibitors, to enhance its safety and digestibility as livestock feed. However, toasting also results in protein denaturation, compromising its solubility and functionality, rendering it unsuitable for high-quality, food-grade applications. Additionally, toasting adversely affects the sensory properties of guar korma, leading to a darker color, oxidized fats, and unfavorable flavors and odors, further limiting its potential for human food applications. (Sandhu, P.P., Bains, K., Singla, G. et al. Nutritional and Functional Properties of Defatted, Debittered and Off-Flavour Free HighVHPM 00773. OOlWOl Protein Guar (Cyamopsis tetragonoloba) Meal Flour. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 89, 695-701 (2019)).

[0009] While cluster bean protein has considerable potential as a nutritional ingredient due to its balanced amino acid profile (Kotnala, Bhavya et al. “Physicochemical, structural, and functional characterization of guar meal protein isolate (Cyamopsis tetragonoloba) ” Heliyon vol. 10,3 e24925. 20 Jan. 2024, doi: 10.1016 / j. heliyon.2024. e24925), it has not been commercialized for human consumption beyond preliminary academic research efforts. For example, M.M. Khalil’s study, Biochemical and Technological Studies on the Production of Isolated Guar Protein (Khalil, M.M. (2001), Biochemical and technological studies on the production of isolated guar protein. Nahrung, 45: 21-24), explored methods for producing isolated guar protein in a laboratory setting. Khalil’s work focused on biochemical extraction techniques, including the use of hexane as a petroleum-based solvent to remove oils during the isolation process.

[0010] However, the process relied on hexane, which is not suitable with recent demand for clean-label, hexane-free processing due to health and environmental concerns associated with petroleum solvents.

[0011] Another example is the series of patents assigned to Shree Ram Proteins Ltd. (e.g., WO 2022 / 144916 Al and related filings), which describe ethanol-based extraction of guar meal for producing protein concentrates. However, these processes rely on (i) an extraction polarity sequence that begins with aqueous or polar ethanol washes before non-polar (e.g., hexane or other solvents) extraction and (ii) desolventization at elevated temperatures. Such conditions cause increased pigment retention, lipid oxidation, protein denaturation, and heat-induced reactions that darken the material and generate off-flavors. The resulting heat-denatured concentrates exhibit reduced solubility, lower emulsification, and weak gelation functionality, rendering them unsuitable for direct human food applications.

[0012] Accordingly, improved processes are needed for extracting food-grade cluster bean protein as a sustainable, clean-label ingredient for incorporation into a variety of food products suitable for human consumption.

[0013] SUMMARY

[0014] In one aspect, provided herein is a method of producing a cluster bean protein concentrate (CPC), comprising:

[0015] (a) providing un-toasted cluster bean germ;

[0016] (b) contacting the germ with a first solvent comprising ethanol of at least 190 proof to separate out a first protein-rich solid by removing at least one of;VHPM 00773. OOlWOl (c) contacting the first protein-rich solid with a second solvent comprising aqueous ethanol of about 120-160 proof to separate out a second protein-rich solid; and (d) desolventizing and drying the second protein-rich solid under vacuum conditions at a temperature below about 75 °C to produce the CPC.

[0017] In one aspect, provided herein is a method of producing a cluster bean protein concentrate (CPC), comprising:

[0018] (a) providing un-toasted cluster bean germ;

[0019] (b) contacting the germ with an aqueous ethanol solvent of about 120-160 proof to produce a protein-rich solid; and

[0020] (c) desolventizing and drying the protein-rich solid under vacuum conditions at a temperature below about 75 °C.

[0021] In one aspect, provided herein is a method of producing a cluster bean protein isolate (CPI), comprising:

[0022] (a) hydrating the CPC at a CPC raw material solids-to-water ratio of about 1 :7- 1:14 to produce a first slurry;

[0023] (b) adjusting the first slurry to a pH of about 7.5-9.5 with a food-grade alkaline agent and applying high-shear mixing at 40-65 °C to produce solubilized proteins;

[0024] (c) separating the solubilized proteins from insoluble fiber to produce a proteinrich liquid;

[0025] (d) acidifying the protein-rich liquid to pH 4.3-4.8 to precipitate a protein solid; (e) resuspending and neutralizing the protein solid to a pH of about 6.0-8.0 to produce a second slurry;

[0026] (f) subjecting the second slurry to direct steam injection at about 120-145 °C for about 5-30 seconds, followed by rapid cooling below about 70 °C, and drying to yield the CPI.

[0027] In one aspect, provided herein is a cluster bean fiber-protein fraction (CFP) comprising the insoluble fiber, wherein the CFP comprises greater than about 25% protein on a dry basis, about 50-60% dietary fiber, and less than about 8% moisture.

[0028] In one aspect, provided herein is a method of producing a functionalized CPC (FCPC), the method comprising hydrating the CPC to produce a slurry, adjusting the pH to about 6-8, subjecting the slurry to direct steam injection at about 120-145 °C for about 5-30 seconds followed by rapid cooling below about 70 °C, and drying the slurry to yield the FCPC.

[0029] In one aspect, provided herein is a method of producing a texturized CPC (TCPC), comprising hydrating the CPC produced by the method of any of claims 1-6 or 9-12 to 10-40%VHPM 00773. OOlWOl moisture, extruding at 80-150 °C using a single- or twin-screw food extruder to form a fibrous expanded product, and drying to less than 10 % moisture.

[0030] In one aspect, provided herein is a method of producing a texturized CFP (TCFP), comprising hydrating the insoluble fiber to 10-40% moisture, extruding at 80-150 °C using a single- or twin-screw food extruder to form a fibrous expanded product, and drying to less than 10 % moisture.

[0031] In one aspect, provided herein is a method of producing high-protein extruded crisps (HPC), the method comprising blending 50-90% CPC with 10-50% starch, adjusting moisture to 10-20 %, extruding in a single- or twin-screw food extruder at 70-150 °C, and drying to 5-8% moisture to yield expanded crisps.

[0032] In one aspect, provided herein is a food product comprising the CPC.

[0033] In one aspect, provided herein is a food product comprising the CPI.

[0034] In one aspect, provided herein is a food product comprising the CFP.

[0035] In one aspect, provided herein is a food product comprising the FCPC.

[0036] In one aspect, provided herein is a food product comprising the TCPC.

[0037] In one aspect, provided herein is a food product comprising the TCFP.

[0038] In one aspect, provided herein is a food product comprising the HPC.

[0039] BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1. Process flow chart of a method for concentrating and isolating food-grade protein and fiber-protein fractions from cluster bean germ (guar korma).

[0041] Figure 2. Continuation of the process flow chart of Figure 1.

[0042] Figure 3. Continuation of the process flow chart of Figure 2.

[0043] Figure 4. Continuation of the process flow chart of Figure 1.

[0044] Figure 5. Continuation of the process flow chart of Figure 1

[0045] Figure 6. Photographic comparison of CPI product prepared by the dual-purity CPC process of Example 4. Left: 1:4 Gel showing firm, elastic structure. Right: 1:4:4 Emulsion showing stable, cohesive emulsion with no oil or water separation.

[0046] Figure 7. Photographic comparison of functional performance of untreated CPC (top) versus FCPC (bottom). Left: 1:4 Gel; Right: 1:4:4 Emulsion. The FCPC exhibits a self-supporting, elastic gel with reduced syneresis and a cohesive emulsion with minimal oil separation, whereas the untreated CPC forms a weak gel and a less stable emulsion.VHPM 00773. OOlWOl DETAILED DESCRIPTION

[0047] The present invention relates to extracting and processing protein concentrates, isolates, and a protein-fiber fraction from cluster beans, also known as Guar (botanical name Cyamopsis tetragonoloba). The disclosed hexane-free, aqueous-ethanol-based process for extracting cluster protein concentrate, followed by a high shear water extraction process, provides a sustainable, green, clean-label technique for producing cluster bean protein suitable for direct incorporation into food products for human consumption. The present invention also employs an optional initial high-purity, non-polar ethanol stage followed by a lower-purity aqueous ethanol stage, combined with low-temperature (< 75 °C) vacuum desolventization and drying. This approach preserves light color (e.g., by avoiding pigment retention, oxidation, and Maillard browning), a bland sensory profile, and functional protein quality while enabling clean-label, food-grade concentrates suitable for industrial production.

[0048] The present invention addresses limitations found in the prior art by implementing a novel, multi-step process that includes pre-cleaning and color sorting of un-toasted cluster bean germ (guar korma); optimized surface area by flaking or milling; a sequential ethanol extraction comprising an initial high-purity, substantially low-polarity ethanol stage (>90% v / v, e.g., 190-200 proof) to remove lipids and pigments, followed by a lower-purity, higher-polarity aqueous ethanol stage (60-80% v / v, 120-160 proof,) to remove saponins, sugars, and other polar impurities, and low-temperature desolventization, drying and milling stages to produce Cluster bean Protein Concentrate (CPC) consist of at least about 82% protein on a dry basis, less than about 4% fat, less than about 10% moisture, and about 3-6% ash; the CPC product is then rehydrated under controlled high-shear alkaline treatments to produce Functional CPC (FCPC), Cluster Bean Protein Isolate (CPI), and a Cluster bean Fiber-Protein mix (CFP) products. These innovations help preserve the protein’s native structure, functional properties, and sensory quality, producing stable, light-colored, mild-flavored protein concentrates and isolates. This approach allows for the production of high-yield and high-quality cluster bean proteins suitable for various food applications.

[0049] The present inventors discovered that a deliberate two-stage ethanol extraction — first with a high-purity, essentially low-polarity ethanol wash (e.g., 190-200 proof), followed by a lower-purity, higher-polarity aqueous ethanol wash (e.g., 120-160 proof) — is critical to obtaining light-colored, bland, food-grade protein from guar korma germ. Unlike prior art (e.g., WO 2022 / 144916 Al (Shree Ram)), which reverses solvent polarity or employs harsher polar solvents under higher heat, this sequence first strips lipids, pigments, and resinous compounds under mild conditions, then selectively removes saponins, sugars, and other polar impurities while preserving protein functionality. This order of operations also enables industrialVHPM 00773. OOlWOl scalability: the first step uses recyclable high -purity ethanol to remove non-polars; the second runs efficiently with aqueous ethanol that is readily recovered and recycled.

[0050] In addition to the solvent polarity sequence, conventional high-heat desolventization (>90°C) causes Maillard browning, protein denaturation, lipid oxidation, and off-flavor development that limit food use. By applying vacuum-assisted low-temperature desolventization and drying (<75 °C, preferably <65 °C) immediately after extraction, the process preserves native protein structure, bland flavor, and light color while preventing oxidation and browning. These qualities make the CPC and derived isolates and fiber fractions more functional and acceptable for food applications at industrial scale.

[0051] The present invention provides a practical solution for unlocking the nutritional potential of the cluster bean germ (guar korma) for human consumption. The disclosed process offers new applications for cluster bean protein in plant-based foods, including Texturization of the CFP and CPC products to be used as textured vegetable proteins (TCFP and TCPC, respectively) in meat and meat-analogs applications or formulated into high-protein extruded crisps by blending with starches or other puffing agents, and the use of the CFP, CPC, FCPC, and CPI in dairy meat analogs, baked goods, high protein plant-based beverages, and other functional food products.

[0052] The present invention provides a novel method for concentrating and isolating foodgrade protein and fiber-protein fractions from cluster bean germ (guar korma), expanding the use of cluster bean seed derivatives beyond animal feed. The process steps are described in detail below in conjunction with Figures 1-5.

[0053] As shown in Figure 1, cluster bean seeds are processed, via milling and sieving, into guar split (guar gum), guar churi (hulls) and non-toasted guar korma which consists mainly the cluster bean germ. While guar korma has traditionally been toasted and incorporated into animal feed, the present process begins with un-toasted cluster bean germ (guar korma), yielding high-quality plant proteins and fiber suitable for human consumption.

[0054] Step l.a. Raw Material: Un-Toasted Cluster Bean Germ (Guar Korma)

[0055] The use of un-toasted cluster bean germ (guar korma) as the raw material is central to producing high-quality food-grade protein and fiber. Un-toasted cluster bean germ, derived from cluster bean seeds after gum extraction, preserves the nutritional profile, including the protein’s native structure, which would otherwise be compromised by toasting. Toasting is a conventional practice to reduce anti -nutritional factors (ANFs) such as trypsin inhibitors, but it denatures the protein, reducing solubility, flavor, and functionality. In contrast, un-toasted cluster bean germ (guar korma) retains the protein's functional properties, making it ideal for human nutrition. The un-toasted cluster bean germ ’s native protein and fiber compositionVHPM 00773. OOlWOl supports the production of high-quality, minimally processed ingredients for human food applications, catering to the increasing consumer demand for sustainable, clean-label food sources.

[0056] Step l.b. Optional Deep Cleaning

[0057] An optional deep cleaning step is employed prior to extraction to significantly improve the quality of the cluster bean germ (guar korma). This stage focuses on physically removing impurities such as residual hulls, rods, and other foreign materials from the germ. The deep cleaning process enhances the appearance, flavor, and sensory characteristics of the final product by eliminating unwanted materials that could affect quality. Additionally, this step contributes to the food safety of the cluster bean germ by selectively removing foreign contaminants, thus ensuring that the raw material is better suited for food-grade applications.

[0058] One key benefit of the deep cleaning stage is the improvement in the protein content of the cluster bean germ (guar korma). Feed-grade guar korma typically contains a wide range of residual hulls and foreign material, with protein content varying from approximately 50% to 60%. By applying this deep cleaning stage, it is possible to standardize the protein purity to a higher range, thereby improving the overall yield and process consistency in the downstream processing stages. This ensures that the cluster bean germ, after deep cleaning, is better suited for food-grade processing and can contribute to more efficient extraction of proteins and fiber fractions.

[0059] Step l.c. Optional Preprocessing: Color Sorting

[0060] An optional color-sorting step improves the quality of the cluster bean germ (guar korma) prior to extraction. This step removes darker or discolored particles that may negatively affect the final product’s appearance, flavor, and sensory characteristics. The color sorting process is performed using advanced optical or digital sorting technologies, such as RGB optical sorters with HD imaging and Al-driven software, which can detect and separate particles based on their hue, brightness, and intensity. Eliminating impurities like oxidized or degraded particles ensures that the final protein and fiber fractions have consistent visual appeal and meet high-quality standards — this quality control benefits premium food applications where sensory attributes such as color and flavor are critical.

[0061] Step l.d. Preparation - Flaking / Coarse Milling / Pelletizing

[0062] In this stage, the cluster bean germ is prepared for efficient extraction. This is typically achieved through conditioning, which involves heating the cluster bean germ to about 40-85°C and adjusting the moisture content to about 10-15%, softening the seed's cell walls. The conditioning process lasts about 30 to about 90 minutes and may involve adding steam or water to ensure uniform moisture distribution. After conditioning, the moist cluster bean germ passesVHPM 00773. OOlWOl through a smooth roller flaker to flatten it into thin flakes (e.g., about 0.1-0.5 mm thick), which increases the surface area and facilitates more effective penetration of the ethanol solution in the subsequent extraction stage. Alternatively, the cluster bean germ can be milled to a particle size of about 200-1,000 microns, improving surface area exposure to the solvent and increasing the extraction efficiency, or pelletized using a low-heat pelletizer. The choice between flaking, milling or pelletizing depends on the specific extraction equipment used and the desired outcome. In some cases, the cluster bean germ may be processed as small grits, which allows for soaking and softening in the solvent prior to milling, optimizing the dissolution and extraction of proteins and other compounds.

[0063] Step l.e. Optional High Purity Ethanol Extraction

[0064] An optional initial extraction step uses high-purity, substantially non-polar ethanol (>90% v / v, preferably 92-96% and up to 200 proof), including specially denatured alcohols (SDA) such as methanol, isopropanol, or n-propanol-denatured ethanol, before any lower-purity aqueous ethanol stage. This non-polar-first sequence efficiently removes lipids, pigments, and other hydrophobic impurities while maintaining protein functionality, and contrasts sharply with prior art such as WO 2022 / 144916 Al, which employs the opposite polarity order and higher heat. The extraction occurs at temperatures between about 20°C and about 60°C, preferably about 40°C to 55°C, more preferably about 50-55 °C (40-55 °C operable) to optimize the removal of solubles, with solvent-to-solid ratios per wash ranging from 10:1 to 2:1 (w / w), preferably 5:1 to 3:1 for efficiency. During extraction, the ethanol primarily removes fat and polar fats, which are undesirable solubles.

[0065] The extraction is typically performed in multiple stages, with fresh ethanol used for successive washing cycles or a counter-current flow design, until at least about 6-12% of the raw material’s dry matter is extracted into the miscella phase. This staged approach ensures maximum removal of soluble impurities without excessive protein loss. Extraction can be performed using various techniques, such as counter-current percolation with equipment like the CROWN Iron Works continuous loop Model II and Model III percolation extractors and / or Model IV immersion extractor, or filtration systems like the Nutsch Filter. High-force decanter centrifugation may also be employed to perform the process.

[0066] Step l.f. Aqueous Ethanol Extraction

[0067] Following the initial high-purity ethanol defatting stage, or, in an alternative embodiment, directly after cleaning and preconditioning — the un-toasted, cleaned cluster bean germ or the partially defatted solids are subjected to a lower-purity aqueous ethanol extraction (60-80% v / v, preferably 65-72%). This stage efficiently removes saponins, sugars, and other polar solubles while retaining proteins and fiber. The process uses a mixture of water andVHPM 00773. OOlWOl ethanol, typically in the range of about 60% to about 80% ethanol (the ethanol may also be provided as SDA containing methanol, isopropanol, or n-propanol) by weight, with an optimal concentration of about 65-72%. The extraction occurs at temperatures between about 20°C and about 60°C, preferably about 40°C to 60°C, to minimize protein denaturation and optimize the removal of solubles and with solvent-to-solid ratios per wash ranging from 10:1 to 2:1 (w / w), preferably 5:1 to 3:1 for efficiency. During extraction, the ethanol causes the protein to temporarily refold and become insoluble, allowing it to remain in the extraction matrix while undesirable solubles are washed away. The extraction is typically performed in multiple stages, with fresh ethanol used for successive washing cycles or a counter-current flow design, until at least about 20-30% of the raw material’s dry matter is extracted into the miscella phase. This staged approach ensures maximum removal of soluble impurities without excessive protein loss, resulting in a purified protein and fiber fraction that is ideal for food-grade applications.

[0068] Extraction can be performed using various techniques such as counter-current percolation with equipment like the CROWN Iron Works continuous loop Model II and Model III percolation extractors and / or Model IV immersion extractor, or filtration systems like the Nutsch Filter. High-force decanter centrifugation may also be employed to separate the solid fraction efficiently, optimizing solvent recovery and increasing overall process efficiency. This stage can be operated batchwise in Nutsche-type filter-dryers or in continuous counter-current immersion / percolation systems to reduce overall solvent-to-solid ratios.

[0069] After extraction, vacuum low-temperature drying (<75 °C, preferably <65 °C) minimizes heat-induced protein denaturation, oxidation and Maillard reactions, keeping the protein lightcolored, bland, and highly functional. This contrasts with high-heat steam stripping, which impairs sensory quality and protein performance in foods.

[0070] Step l.g. Low- Temperature Desolventization and Drying of Cluster Bean Protein Concentrate (CPC)

[0071] After the aqueous ethanol extraction, the residual protein-fiber solids, now in the form of an insoluble cake, undergo a pressing stage to remove any residual liquids. This can be achieved using a Dewatering Screw Press (e.g., Vincent-Type press). Alternatively, if a decanter centrifuge or batchwise Nutsche-type filter-dryers is used, the cake’s moisture content may already be sufficiently reduced, and additional pressing may not be required. The semi-dry cake is then subjected to a low-temperature drying process to achieve a final product with less than about 12% moisture content.

[0072] The Low-temperature desolventization is performed under vacuum at product temperatures maintained below 75 °C, preferably below 65 °C. These conditions minimize lipid oxidation, heat-induced protein denaturation, pigment retention, and Maillard-type browningVHPM 00773. OOlWOl reactions that can darken color and create off-flavors, while preserving protein solubility, emulsification, and gelation functionality. Maintaining low temperature is critical to preserve bland sensory characteristics and protein functionality required for human food applications.

[0073] The drying process can be carried out using systems such as the Crown Iron Works Vacuum Desolventizer Stripper, which operates under vacuum conditions to remove ethanol and other volatiles while minimizing protein degradation. This process can also be performed using Nutsch-type vacuum dryers or other suitable drying technologies, ensuring the product’s drying temperature remains below about 75°C, ideally under about 65°C, to prevent protein denaturation and maintain the quality of the final product.

[0074] The final dried cluster bean Protein Concentrate (CPC) typically contains 80-90% protein (dry basis), with fat <3.5% for dual -purity CPC and less than about 6% for single-purity CPC on a dry matter basis, making it a high-protein, functional ingredient suitable for a range of food formulations.

[0075] Step l.h. Optional Milling

[0076] Once the CPC is dried, it may undergo a further milling stage to reduce its particle size to between about 40 and about 500 microns. This step enhances its use in food applications or other downstream processing, providing a versatile ingredient with improved dispersibility. Step l.i. Food Applications and Usages of CPC

[0077] The dry CPC is characterized by its protein content (> about 80% on a dry basis), mild flavor, light color, and low levels of anti -nutritional factors (ANFs), making it suitable for a wide range of food applications. The CPC can be used directly in baked goods, beverage powders, ready-to-drink shakes, and high-protein nutrition bars. It also serves as the feedstock for Functional CPC (FCPC), Cluster Bean Protein Isolate (CPI), Textured CPC (TCPC), and high-protein crisps (HPC). Its neutral flavor and clean label properties make it ideal for protein fortification in beverages, baked goods, nutritional supplements and bars, meat analogs, and various other food products.

[0078] Since the protein in the CPC is temporarily insoluble, further functionalization can be performed by rehydrating the CPC and adjusting the pH. The CPC may also be treated with crosslinking agents, such as phosphates, or enzymes to improve texture, viscosity, emulsification, and gelation. Once functionalized, the CPC can be pasteurized and spray-dried, with optional particle agglomeration to enhance dispersibility and reduce dustiness. As shown in Figure 2, the process can be continued beginning with the CPC produced in Figure 1.

[0079] Step 2.a. High Shear Alkaline Extraction

[0080] Following the aqueous ethanol extraction, high shear alkaline extraction is the next step in isolating the cluster bean Protein Concentrate (CPC) into cluster bean Protein Isolate (CPI).VHPM 00773. OOlWOl This secondary extraction stage is crucial for purifying and solubilizing the protein while separating it from the remaining insoluble fiber fraction. The CPC is mixed with water at a ratio of about 1 :7 to about 1 : 14, ideally between about 1 :9 to about 1 : 12. An alkaline agent, such as sodium hydroxide (NaOH) or calcium hydroxide (CaOH), is added to adjust the slurry's pH to a range of about 7.5 to about 9.5, with an optimal pH range of about 8.0 to about 9.0. This alkaline environment enhances the protein’s solubility and promotes protein separation from insoluble fiber.

[0081] High-shear mixing uses industrial high-shear mixers with rotor-stator mechanisms to break down fibrous structures and maximize protein solubilization. The mixing conditions, including speed and duration, are carefully controlled to ensure maximum protein release. The temperature is maintained between about 40°C and about 65°C, preferably around about 45°C to about 55°C, to enhance protein solubility while preventing heat-induced denaturation. The mixing typically lasts between about 30 minutes to about 3 hours, with an optimal range of about 60 minutes to about 2 hours to achieve maximum protein yield and solubility.

[0082] The slurry is then fed to a horizontal decanter centrifuge and / or high-speed disc clarifier centrifuge to separate the liquid fraction from the solid insoluble fiber-protein material.

[0083] Following the primary high-shear alkaline extraction, the fiber solids, which may still contain some residual protein, undergo a secondary high-shear alkaline extraction to ensure maximum protein recovery. In this secondary extraction, water is reintroduced to the fiber fraction at a dilution rate of about 1 :4-l : 10 based on CPC raw material solids to water, preferably 1 :6-l :8. This secondary extraction is subject to high shear mixing and pH adjustment in the same way as the first extraction is done, then held for about 30 to about 120 minutes, allowing for further disruption of the fiber matrix and ensuring additional protein recovery. After this, another solid-liquid separation is performed using a horizontal decanter centrifuge and / or high-speed disc clarifier centrifuge.

[0084] A third extraction stage can be done by re-dilution of the secondary fiber fraction at a dilution rate of about 1:4-1:10 based on CPC raw material solids to water, preferably 1 :6-l :8. This third extraction is subject to high shear mixing and pH adjustment in the same way as the first and second extraction stages, then held for about 30 to about 120 minutes, allowing for further disruption of the fiber matrix and ensuring additional protein recovery. After this, another solid-liquid separation is performed using a horizontal decanter centrifuge and / or highspeed disc clarifier centrifuge.

[0085] The liquid fractions from both the primary, secondary and optional third extractions are either collected together for further processing or, in the case of counter-current extraction, the liquids from the second and third extractions are used in the front end of the first and secondVHPM 00773. OOlWOl extractions respectively, and only the more concentrated liquid from the first stage is then collected for further processing. This multi-stage, high-shear alkaline extraction process ensures maximum protein recovery and purity, enhancing the yield of high-quality, functional protein that can be used to produce cluster bean Protein Isolate (CPI)

[0086] Step 2.b. Protein Precipitation

[0087] Protein precipitation is a vital step in isolating and concentrating cluster bean proteins from the alkaline extraction solution. This process involves adjusting the pH of the solution to the isoelectric point of the cluster bean proteins, which is the pH range where the proteins are least soluble and thus precipitate from the solution. The pH is carefully lowered to a range of approximately 4.3 to 4.8 using food-grade acids such as citric acid, hydrochloric acid, or phosphoric acid. At this isoelectric pH, the cluster bean proteins lose their net charge, and promoting aggregation into stable protein particles.

[0088] Once the pH has been adjusted, the slurry undergoes an aging phase, where protein molecules reorganize into stronger, more stable aggregates. This aging phase is essential for improving the cohesion, size, and density of the precipitate, making it easier to separate in the subsequent centrifugation step. During aging, the slurry is gently stirred, allowing the protein aggregates to form more stable structures that settle more effectively when separated.

[0089] After aging, the slurry is transferred to a horizontal decanter centrifuge and / or a high-efficiency disc type clarifier centrifuge . The centrifugation process, which operates at high G-forces, separates the solid protein precipitate from the supernatant (liquid phase). The supernatant contains any remaining soluble sugars, residual proteins, minerals and other compounds, which may be set aside for further processing or potential reuse. The collected protein precipitate represents the isolated cluster bean protein, which is then ready for neutralization and functionalization to optimize its properties for food-grade applications.

[0090] Step 2.c. Neutralization, Functionalization, and Pasteurization of Protein Isolate After either the precipitation step (Step 2.c.) or, in the case of producing Functional CPC after hydration of the CPC (Step l.i.), the CPC or CPI protein curd or solids undergoes hydration / resuspension and neutralization to adjust the pH to a neutral or slightly alkaline level, tailored for the specific functionality requirements of the intended food application. Food-grade alkaline agents, such as sodium hydroxide (NaOH), calcium hydroxide (CaOH), or other approved bases, adjust the pH to a range of about 6.0 to about 8.0, depending on the target application.

[0091] Following neutralization, the protein undergoes functionalization to enhance its properties for diverse food applications. Various functionalization treatments can be employed, including:VHPM 00773. OOlWOl • Enzyme Reactions: Enzymatic modification can hydrolyze the protein into smaller peptide sizes, improving solubility, digestibility, emulsification, and foaming properties. This enhances the protein’s behavior in formulations requiring specific textures or stability, such as protein beverages and whipped products.

[0092] • Crosslinking: Crosslinking agents, such as transglutaminase and other binding agents, can strengthen the protein’s molecular structure, improving gel strength for applications such as egg albumin replacement or plant-based meat analogs.

[0093] • High-Pressure Homogenization: High-pressure homogenization can further improve the smoothness, gel strength, and emulsification stability of the protein, ensuring uniform particle size and dispersion. This enhances the texture and mouthfeel in dairy alternatives, protein beverages, sauces, and other food products.

[0094] After functionalization, the FCPC or CPI undergoes pasteurization to ensure food safety. High-Temperature Short Time (HTST) pasteurization at temperatures ranging from about 120°C to about 145°C is typically employed. In this process, the protein slurry is rapidly heated to the specified temperature using direct steam injection and held for a duration of about 5 seconds to 30 seconds to eliminate microbial contaminants. Following this, the slurry is quickly cooled down to a temperature range of about 40°C to about 70°C, preferably between about 50°C and about 60°C, to prevent further protein denaturation. This pasteurization process ensures the product's safety while preserving its functionality, texture, and flavor.

[0095] Step 2.d. Drying of CPI / FCPC

[0096] Once pasteurized, the FCPC or CPI is converted into a powdered form suitable for various applications. Spray drying is the preferred method for this process (other drying methods can also be used, such as vacuum drying, freeze drying, fluid-bed drying, and ring drying, for example), where the protein slurry is atomized into a heated chamber, allowing rapid moisture evaporation and producing a fine powder. Additional treatments, such as coating and agglomeration, can be applied after drying to address specific product needs. The coating can reduce dustiness and improve flowability, while agglomeration enhances solubility and functionality for different applications. This versatility makes the FCPC and CPI suitable for a wide range of food and beverage applications, such as protein-enriched formulations, dairy and meat analogs, and nutritional supplements.

[0097] Step 2.e. Food Applications and Uses of FCPC

[0098] FCPC, characterized by its high protein content (> about 80% on a dry basis), is an ideal functional ingredient for high-protein foods, meat analogs, sports nutrition products, and mealVHPM 00773. OOlWOl replacements. It offers a competitive alternative to other plant-based proteins. Its mild flavor and light color make it particularly advantageous for formulations where protein and fiber enhancement are desired without altering the product's sensory characteristics.

[0099] Step 2.f. Food Applications and Uses of CPI

[0100] The CPI, characterized by its rich essential amino acid profile and high protein content (> about 90%), is an ideal functional ingredient for high-protein foods, plant-based beverages, non-dairy yogurts, sports nutrition products, and meal replacements. CPI exhibits superior emulsification and viscosity properties, offering a competitive alternative to other plant-based proteins. Its versatility enhances its appeal across various food applications.

[0101] Step 2.g. Optional Fiber Pre-Treatments

[0102] The drying process for the CFP is crucial to achieving a high-quality, food-grade product that meets specific functional and nutritional requirements. Various pre-treatment techniques may be applied to optimize the quality and stability of the CFP before drying. These pretreatments help the CFP meet the demands of sensory, nutritional, and functional performance in different food applications. In certain embodiments, pre-treatment techniques prior to drying can include one or more of the following:

[0103] Heat Treatment for Micro- and Nutrient-Stabilization:

[0104] A heat treatment, typically in the range of about 72°C to about 135°C for a period of about 5 seconds to about 1 hour, stabilizes nutrients and reduces microbial load. This treatment improves the solubility and digestibility of the fiber, making it suitable for food-grade applications while preserving the integrity of the fiber-protein fraction.

[0105] Enzymatic Cleavage for Prebiotic Fiber Production:

[0106] Enzymatic treatment, such as the use of Cell Wall Degrading Enzyme Complexes (e.g., Viscozyme® L, which includes beta-glucanases, pectinases, hemicellulases, and xylanases), selectively breaks down fiber components to produce prebiotic fibers. These prebiotic fibers can enhance digestive health, adding value to the CFP for functional food products targeting gut health.

[0107] Chemical Treatments to Modify Fiber Solubility:

[0108] Aqueous or mild chemical treatments, such as hydrogen peroxide, may be applied to modify the solubility and functional properties of the fiber. This enhances hydration properties, making the fiber more versatile for applications requiring specific water-holding capacity.

[0109] Step 2.h. Low Temperature Drying

[0110] Once pre-treatments are applied, low-temperature drying methods are employed to ensure that the product maintains its quality and stability while preserving its color, flavor, andVHPM 00773. OOlWOl nutritional profile. In certain embodiments, low-temperature drying methods include one or more of the following:

[0111] Spray Drying:

[0112] Spray drying is used to rapidly dry the CFP fraction while preserving its functional and sensory qualities. This method is effective for producing uniform, high-quality powders suitable for a variety of food applications.

[0113] Ring Dryer:

[0114] Ring dryers use indirect heating and efficient air circulation to dry the CFP fraction, helping to preserve its color and flavor. The gentle heating conditions prevent browning, and the controlled drying process ensures nutrient retention, making ring drying ideal for producing functional, nutritionally valuable ingredients.

[0115] Fluidized Bed Dryer:

[0116] This method uses an upward airflow to ensure even drying while maintaining the structural integrity of the protein and fiber components. It is particularly suitable for producing consistent, high-quality products that require a light color and mild flavor.

[0117] Air-Swirl Dryer:

[0118] The air-swirl dryer employs centrifugal force combined with heated air to minimize direct heat exposure, helping to retain the natural color and nutritional profile of the CFP. This method is particularly well-suited for processing sensitive materials and ensuring a high-quality, consistent dried product.

[0119] By combining these pre-treatment and drying technologies, the CFP fraction achieves superior food-grade quality, enhanced functionality, and versatility. The resulting product can be used in a wide range of applications, including health-oriented foods, bakery items, meat and dairy analogs, and high-fiber applications, making it a valuable ingredient for diverse food formulations.

[0120] Step 2.i. Food Applications and Uses of CFP

[0121] This invention highlights the versatile applications of the CFP fraction within the food industry, showcasing its novel uses in human-grade products. CFP's unique properties position it as a valuable ingredient for a wide range of food applications, addressing the growing demand for plant-based proteins and functional fibers.

[0122] Fiber-Enhanced Applications:

[0123] The CFP, containing about 40-50% protein and about 25-35% total dietary fiber, offers an optimal nutritional balance for health-focused products. The dietary fiber in CFP supports digestive health, making it a valuable ingredient in products such as bakery items, cereals, snacks, and beverages targeting dietary fiber claims. Additionally, the high fiber contentVHPM 00773. OOlWOl contributes to structure and moisture retention, which is particularly beneficial in gluten-free baking and other texture-sensitive food applications.

[0124] Health and Clean Label Benefits:

[0125] CFP is naturally free from common allergens such as gluten and soy, making it highly compatible with the clean-label trend in food manufacturing. This absence of significant allergens increases the appeal of cluster bean-based ingredients in products designed for consumers with dietary sensitivities, including gluten intolerance. As a result, CFP offers opportunities for inclusion in allergen-free and specialty food products, catering to the needs of health-conscious and allergen-sensitive consumers.

[0126] As shown in Figure 3, the process can be continued beginning with the CFP produced in Figure 2. The processing of Textured Cluster Fiber Protein (TCFP) begins with a preconditioning stage, where water or steam is added to the protein powder (CFP) to adjust its moisture content to 10-40%. At this stage, other additives, such as food-grade colors, flavors, proteins, starches, and oils, can be incorporated based on the desired product outcome.

[0127] Once the pre-conditioning is complete, the material is fed into an extruder. The extrusion process can use either a single or twin-screw extruder, depending on the product requirements. A twin-screw extruder offers better control of mixing and processing, while a single-screw extruder is often simpler and more economical. The extruder works by forcing the conditioned material through a die, which can be designed in various shapes and sizes based on the finished product's requirements.

[0128] Following extrusion, the material is cut into the desired shapes using shredders, slicers, or dicers. Machines such as the ones from Urschel®, designed for cutting extruded foods, are used to slice or dice the extruded product into uniform pieces, depending on the specifications of the finished product.

[0129] Finally, the product is dried to achieve the desired moisture content of 7-12%, which is ideal for storage and product stability. This drying process can be carried out using multi-stage dryers or fluidized bed dryers, which ensure uniform heat distribution and effective moisture removal without compromising the product’s quality.

[0130] This process ensures the production of high-quality, consistent TCFP, suitable for a wide variety of food applications, from meat analogs to plant-based snacks and protein-enriched formulations.

[0131] Step 3.a. Food Applications and Uses of CFP as Raw Material for Textured Cluster bean germ Protein (TCP) Production

[0132] The CFP derived from guar korma can be processed into Textured Cluster bean germ Protein (TCP), a high-protein (>45%), high-fiber (>25%) ingredient ideal for meat analogs,VHPM 00773. OOlWOl plant-based burgers, and other food products where texture replication is critical. Through processes such as extrusion and other texturizing techniques, TCP achieves a fibrous, meat-like structure. Extrusion forces the CFP through a die under high temperature and pressure, which aligns the protein and fiber components to form a texture similar to that of animal-based meat. Additional texturizing techniques, such as High Moisture Extrusion (EIME) or dry spinning, may be used to refine the texture and improve the product's final mouthfeel. This development supports the growing alternative protein market, providing a plant-based meat substitute with a desirable texture and nutritional profile.

[0133] As shown in Figure 4, the process can continue beginning with the CPC produced in Figure 1. The processing of Textured Cluster Bean Protein Concentrate (TCPC) begins with a pre-conditioning stage, where water or steam is added to the protein powder (CPC) to adjust its moisture content to 10-40%. At this stage, other additives, such as food-grade colors, flavors, proteins, starches, and oils, can be incorporated based on the desired product outcome.

[0134] Once the pre-conditioning is complete, the material is fed into an extruder. The extrusion process can use either a single or twin-screw extruder, depending on the product requirements. A twin-screw extruder offers better control of mixing and processing, while a single-screw extruder is often simpler and more economical. The extruder works by forcing the conditioned material through a die, which can be designed in various shapes and sizes based on the finished product's requirements.

[0135] Following extrusion, the material is cut into the desired shapes using shredders, slicers, or dicers. Machines such as the ones from Urschel®, designed for cutting extruded foods, are used to slice or dice the extruded product into uniform pieces, depending on the specifications of the finished product.

[0136] Finally, the product is dried to achieve the desired moisture content of 7-12%, which is ideal for storage and product stability. This drying process can be carried out using multi-stage dryers or fluidized bed dryers, which ensure uniform heat distribution and effective moisture removal without compromising the product’s quality.

[0137] This process ensures the production of high-quality, consistent TCPC, suitable for a wide variety of food applications, from meat analogs to plant-based snacks and protein-enriched formulations.

[0138] Step 4.a. Food Applications and Uses of CPC as Raw Material for Textured Cluster Bean Protein Concentrate (TCPC) Production

[0139] The CPC derived from guar korma can be processed into Textured Cluster bean germ Protein Concentrate (TCPC), a high-protein (>80% on a dry basis) and high-fiber (>9%) ingredient suitable for meat analogs, plant-based burgers, and other food products where textureVHPM 00773. OOlWOl replication is critical. TCPC achieves the desired fibrous, meat-like structure through extrusion and other texturizing techniques. Extrusion involves forcing the CPC through a die under high temperature and pressure, which aligns the protein and fiber components to create a texture resembling animal-based meat. Additional texturizing methods such as High Moisture Extrusion (HME) or dry spinning may also be employed to refine the texture and improve the final mouthfeel of the product. This advancement supports the growing alternative protein market, offering a plant-based meat substitute with a desirable texture and nutritional profile.

[0140] As shown in Figure 5, the process can continue beginning with the CPC produced in Figure 1. The production of High -Protein Crisps (HPC) begins with a pre-conditioning stage, where the CPC is blended with starches or other puffing agents (e.g., com / potato tapioca starch) to adjust composition and water content to about 10%-40%. Water or steam may be added to reach the target feed moisture. At this stage, other food-grade ingredients, such as colors, flavors, or oils, can be incorporated depending on the desired sensory and nutritional profile.

[0141] Once pre-conditioning is complete, the material is fed into an extruder (single- or twin-screw, depending on scale and control requirements). The extrusion step subjects the mixture to controlled heat and shear to promote expansion and crisp structure. The extruder forces the conditioned material through a die that determines the final shape and size of the crisps.

[0142] Following extrusion, the expanded strands are cut to size using cutting devices such as rotary cutters or dicing blades (e.g., Urschel® cutters). The extruded pieces are then dried to achieve a final moisture content of about 5-10%, providing shelf stability and crisp texture. Drying can be performed using multi-stage hot air dryers or fluidized-bed systems to ensure even heat distribution and preserve the light color and mild flavor of the product.

[0143] This process yields high-protein crisps (HPC) with a light, , less than 4 % fat, less than 10 % moisture airy, crunchy texture and excellent rehydration and flavor performance. HPC serves as a clean-label, high-protein snack or inclusion for bars, cereals, and ready-to-eat applications. Step 5.a. Food Applications and Uses of CPC for High-Protein Crisps (HPC) Production CPC-based HPC, with protein contents typically above 50% (dry basis), offers a versatile, allergen-friendly alternative to soy- or pea-based crisps. These crisps can be used as high-protein snacks, inclusions in nutrition bars, breakfast cereals, or toppings for plant-based meals. The mild flavor, light color, and clean-label profile of CPC-based crisps make them particularly attractive for formulating premium plant-protein snacks and functional foods. As used herein, the terms “guar bean” and “cluster bean” refer to Cyamopsis lelragonoloba the terms are used interchangeably herein.

[0144] As used herein, in certain aspects, the term “comprising” can be substituted with the term “consisting of,” or “consisting essentially of’ with respect to the invention.VHPM 00773. OOlWOl The invention will now be illustrated by the following non-limiting Examples.

[0145] EXAMPLES

[0146] While the following examples describe CPC preparation using a batch-operated Nutsche-type filter-dryer, the same sequence of high-purity followed by a low-purity ethanol extractions can also be performed in continuous counter-current extraction systems. In such systems, equivalent removal of non-polar and polar impurities is achieved, while reducing solvent-to-solid ratios and improving solvent recovery efficiency. The counter-current configuration is therefore considered within the scope of the described methods and is applicable to each of the CPC extraction examples provided below.

[0147] Example 1: Preparation of Cluster Bean Protein Concentrate (CPC) by Dual-Purity Ethanol Extraction

[0148] Un-toasted guar korma germ (supplied by Manish Agro, India) was sieved to remove hulls, stems, and other foreign matter, and passed through a color sorter to remove dark particles. This step reduced polyphenol and pigment load, contributing to a cream-colored final product. The cleaned germ was milled in a Fitzpatrick Fitzmill JT6 hammer mill, a compact pilot-scale unit equipped with a #1050 perforated plate, producing a coarse flour. The flour was characterized by 8.6-9.0% moisture, 61.0% protein (dry basis), and 6.0% fat.

[0149] The prepared flour was charged into a Nutsche-type filter-dryer manufactured by Paul Mueller Company (0.2 m2filtration area, S / N 142886, built 1995). The vessel was constructed of 316L stainless steel with a 304 stainless steel jacket and rated for 90 PSIG and full vacuum at 350 °F. It was equipped with a 150 pm perforated filter plate at the base and an internal agitator for slurry mixing, cake smoothing, and displacement washing. The jacketed design enabled both heating and cooling, and the closed system allowed solvent contact, washing, solid-liquid separation, and vacuum drying within the same equipment. Miscella displacement was assisted by applying nitrogen head pressure (10-45 psi). Although the unit used here was a 0.2 m2pilot system, the geometry is directly scalable to industrial Nutsche filter-dryers with filtration areas of 2-20 m2or larger.

[0150] High-purity ethanol stage

[0151] The flour was contacted with 200-proof ethanol under slow agitation at 50 °C. Two successive washes were carried out at solvent-to-solid ratios in the range of 5: 1 to 3.5: 1 (w / w). After each contact period, the miscella was displaced through the filter by applying nitrogen head pressure. This stage removed non-polar lipids, pigments, and resinous impurities.

[0152] Low-purity aqueous ethanol stageVHPM 00773. OOlWOl The partially defatted solids were then contacted sequentially with 140-proof ethanol at 52-55 °C. Three successive washes were performed at solvent-to-solid ratios of 4:1 to 3:1 (w / w), with nitrogen pressure applied after each to expel the miscella. This stage removed residual saponins, sugars, and polar solubles while preserving protein functionality.

[0153] Desolventization and drying

[0154] The washed solids were desolventized and dried in the same Nutsche-type filter-dryer under -24 inHg vacuum, with the jacket temperature maintained below 75 °C. Drying was continued until the product reached <10% final moisture. These mild drying conditions were key to preventing off-flavor and color darkening, resulting in food-grade CPC suitable for beverages, bars, high protein crisps and meat analogues. The dried CPC was subsequently milled in a Prater Sterling Rotomill 1300 operating at 50 Hz to a particle size of D(90) - 180 pm, yielding a fine, uniform powder.

[0155] Product

[0156] The resulting CPC was cream-colored, flavorless, odorless (per internal sensory panel as defined in Analytical Methods), and readily dispersible in water, with 90% protein (dry basis), 8-9% moisture, 0.35-0.78% fat, and 4.67% ash. This outcome was reproducible across replicate runs using the same method, consistently producing CPC with >84% protein (dry basis) and <1% fat.

[0157] Example 2: Preparation of Cluster Bean Protein Concentrate (CPC) by Dual-Purity Ethanol Extraction Using Industrial-Grade Solvent

[0158] Un-toasted guar korma germ (supplied by Manish Agro, India) was sieved to remove hulls, stems, and other debris, and passed through a color sorter to eliminate darker particles. The cleaned germ was conditioned to -12% moisture by controlled water addition and tumble mixing, then pre-warmed at -37 °C under -70% relative humidity for 40 minutes. The tempered germ was flaked on a Creason smooth roller mill adjusted to a 0.006" (0.152 mm) gap, producing thin, uniform flakes. The flakes were subsequently dried with warm air at -37 °C to reduce the moisture content to -9.5%. Proximate analysis of the flakes confirmed 9.55% moisture, 61.0% protein (dry basis), and 6.0% fat.

[0159] The flakes were extracted in a Nutsche-type filter-dryer manufactured by Paul Mueller Company (0.2 m2, S / N 142886) as described in Example 1. In this run, the high-purity stage used recovered anhydrous ethanol at approximately 195 proof, reflecting the practical solvent purity achievable in industrial recovery systems. In contrast, Example 1 employed 200-proof reagent ethanol, which requires a more robust anhydrous ethanol recovery system.

[0160] High-purity ethanol stageVHPM 00773. OOlWOl The flakes were contacted with -195-proof ethanol at 50-60 °C under slow agitation. Two successive washes were carried out at solvent-to-solid ratios in the range of 5: 1 to 3.5: 1 (w / w), with miscella displaced by nitrogen pressure after each wash. This stage effectively removed non-polar lipids and pigments, ensuring a clean sensory profile.

[0161] Low-purity aqueous ethanol stage

[0162] The partially defatted solids were then contacted sequentially with 140-proof ethanol at -52 °C. Three successive washes were performed at solvent-to-solid ratios of 4:1 to 3:1 (w / w), with miscella displaced after each wash. This stage further reduced saponins, sugars, and polar solubles.

[0163] Desolventization and drying

[0164] The washed solids were desolventized and dried in the same Nutsche-type filter-dryer under -24 inHg vacuum, with the jacket temperature maintained below 75 °C. Drying was continued until the product reached <10% final moisture (-3.7% in this run). These mild drying conditions were key to preventing off-flavor and color darkening, resulting in food-grade CPC suitable for beverages, bars, high protein crisps, and meat analogues. The dried CPC was subsequently milled in a Prater Sterling Rotomill 1300 operating at 50 Hz to a particle size of D(90) - 180 pm.

[0165] Product

[0166] The resulting CPC was cream-colored, dispersible in water, and flavor-neutral (per internal sensory panel as defined in Analytical Methods), with 85.0% protein (dry basis), 3.7% moisture, 3.2% fat, and 4.9% ash. This outcome was reproducible across replicate runs using the same method, consistently producing CPC with >83% protein (dry basis) and <3.5% fat.

[0167] Example 3: Preparation of Cluster Bean Protein Concentrate (CPC) by Single-Purity Ethanol Extraction

[0168] Flakes of guar korma germ, prepared as described in Example 2 (conditioning to -12% moisture, pre-warming, flaking on a Creason smooth roller mill at 0.006" gap, and drying to -9.5% moisture), were used as the feedstock. These flakes were characterized by 9.55% moisture, 61.0% protein (dry basis), and 6.0% fat.

[0169] The flakes were extracted in a Nutsche-type filter-dryer manufactured by Paul Mueller Company (0.2 m2, S / N 142886) as described in Example 1. In contrast to Examples 1 and 2, which included a high-purity ethanol stage, this process employed only a low-purity 140-proof aqueous ethanol for extraction.

[0170] Single-purity ethanol extractionVHPM 00773. OOlWOl The flakes were contacted sequentially with 140-proof ethanol at ~52 °C under slow agitation. Four successive washes were carried out at solvent-to-solid ratios in the range of 6: 1 to 3.5:1 (w / w), with miscella displaced by nitrogen pressure after each wash. This sequence effectively removed saponins, sugars, and polar solubles, but left more residual fat compared to dual-purity processes.

[0171] Desolventization and drying

[0172] The washed solids were desolventized and dried in the same Nutsche-type filter-dryer under -24 inHg vacuum, with the jacket temperature maintained below 75 °C. Drying was continued until the product reached <10% final moisture (-7.7% in this run). The dried CPC was subsequently milled in a Prater Sterling Rotomill 1300 operating at 50 Hz to a particle size of D(90) - 180 pm.

[0173] Product

[0174] The resulting CPC was cream-colored and dispersible in water, with 79.9% protein (dry basis), 7.7% moisture, 4.8% fat, and 5.6% ash. It exhibited a slight residual fatty note in flavor (per internal sensory panel as defined in Analytical Methods), which could limit its inclusion rate or require flavor masking in sensitive applications compared to dual-purity CPC. Although this single-stage approach yields slightly higher residual fat and less neutral sensory profile than the dual-stage process, the controlled low-temperature drying maintains sufficient functionality and clean-label appeal for certain applications. This outcome was reproducible across replicate runs using the same method, consistently producing CPC with >80% protein (dry basis) and <6% fat.

[0175] Example 4: Preparation of Cluster Bean Protein Isolate (CPI) and Cluster Bean Fiber -Protein Fraction (CFP) from Dual-Purity CPC (Pilot Scale)

[0176] Feed material

[0177] Cluster bean protein concentrate (CPC) was prepared by dual-purity ethanol extraction as described in Example 2. The CPC was cream-colored, bland in flavor, and dispersible in water, with 86.9% protein (dry basis), 6.53% moisture, 0.26-0.49% fat, and -4-5% ash.

[0178] Alkaline extraction

[0179] A batch of 8.4 kg CPC was hydrated with water at a solids-to-water ratio of -1 : 12.5 (w / w). The slurry was adjusted to pH 8.5-9.0 with sodium hydroxide and mixed at -50 °C using a high-shear rotor-stator mixer until proteins were fully solubilized. The slurry was then fed into a Tolhurst® Center-Siu basket centrifuge (20-inch model, S / N TA-26115, Ametek Inc., East Moline, Illinois), fitted with a 1 pm food-grade filter bag. The centrifuge was operated at 1690 RPM, generating centrifugal forces of approximately 700 * g, thereby separating the clearVHPM 00773. OOlWOl protein-rich liquids from insoluble solids.. This extraction sequence was repeated two additional times at water ratios between 1:8 and 1 : 12 and temperatures of 50-55 °C. The liquid phases from all three stages were combined for further processing, while the insoluble solids were retained for CFP production.

[0180] Isoelectric precipitation

[0181] The combined alkaline extracts were acidified to pH 4.2-4.8 with hydrochloric acid or citric acid at ~40 °C to precipitate the proteins. The slurry was aged briefly under mild agitation and centrifuged in the same basket centrifuge to recover the protein curd, while the supernatant was discarded.

[0182] Neutralization, Functionalization, and Pasteurization

[0183] The protein curd was resuspended in water with high-shear mixing, neutralized to pH 7.0 with sodium hydroxide, and pasteurized by direct steam injection at 130 °C for 5-10 seconds, followed by rapid cooling in a vacuum chamber to -55 °C.

[0184] Spray drying

[0185] The neutralized slurry was spray dried in a tail-form dryer equipped with a high-pressure nozzle (operated at -4000 psi; inlet 187 °C, outlet 92 °C), yielding a fine powder with <6% moisture.

[0186] CPI Product

[0187] The resulting CPI was light in color, bland in flavor, and readily dispersible in water, with >96% protein (dry basis), 3.24% moisture, <0.4% fat, and 4.38% ash. The isolate demonstrated excellent solubility, emulsification capacity, foaming, and gelation properties, making it suitable for dairy alternatives, plant-based beverages, and high-protein food applications. Comparable results were obtained in repeat pilot-scale runs, consistently producing CPI with >94% protein (dry basis) and <0.5% fat. Representative functionality of a CPI produced according to Example 4 is shown in Figure 6, which was evaluated in the 1 :4 Gel and 1:4:4 Emulsion tests (see Analytical Methods). The gel exhibited a firm, elastic texture resistant to fracture, while the emulsion was cohesive and stable with no oiling-off or water separation after hot-cold processing.

[0188] CFP Product

[0189] The insoluble solids separated during alkaline extraction were freeze-dried and milled to a flour with particle size <250 pm. The CPF comprised -28% protein (dry basis), -60% dietary fiber, <2% fat, and -5% ash, with <8% moisture. The CFP was suitable for use as a flour in baked goods, pasta, gluten-free cookies, and as a raw material for producing texturized vegetable proteins (TVP / TCPC).VHPM 00773. OOlWOl Example 5: Preparation of Cluster Bean Protein Isolate (CPI) and Cluster Bean Fiber-Protein Fraction (CFP) from Single-Purity CPC (Pilot Scale)

[0190] Feed material

[0191] Cluster bean protein concentrate (CPC) was prepared by single-purity aqueous ethanol extraction as described in Example 3. The CPC was cream-colored and dispersible, with 79% protein (dry basis), 8.83% moisture, 5.30% fat, and 4.56% ash.

[0192] Alkaline extraction

[0193] 13.5 kg of CPC was hydrated with water at a solids-to-water ratio of -1:11.5 (w / w). The slurry was adjusted to pH 8.5-9.0 with sodium hydroxide and mixed at -50-55 °C using a high-shear rotor-stator mixer until proteins were solubilized. The slurry was clarified in the same basket centrifuge (Tolhurst® Center-Siu basket centrifuge 20-inch model, S / N TA-26115, Ametek Inc., East Moline, Illinois) described in Example 4. This extraction was repeated twice at water ratios between 1 :6.7 and 1:10 and temperatures of 50-55 °C. The clarified liquids were combined for precipitation, while insoluble solids were retained for CFP production.

[0194] Isoelectric precipitation

[0195] The combined extracts were acidified to pH 4.2-4.8 with hydrochloric acid or citric acid under mild agitation at -40 °C, aged briefly, and centrifuged (basket centrifuge, Tolhurst® Center-Siu basket centrifuge 20-inch model, S / N TA-26115, Ametek Inc., East Moline, Illinois) to collect the protein curd. The supernatant was discarded.

[0196] Neutralization, Functionalization, Pasteurization, and spray drying

[0197] The protein curd was resuspended in water with high-shear mixing, neutralized to pH 7.0 with sodium hydroxide, and pasteurized by direct steam injection at -130 °C for 5-10 seconds in a holding tube, followed by flash vacuum cooling to -55 °C. The neutralized slurry was then spray dried in a tall -form dryer equipped with a high-pressure nozzle (-4000 psi; inlet 186 °C, outlet 91 °C), producing a pale yellow powder with <6% moisture.

[0198] CPI Product

[0199] The resulting CPI contained 87% protein (dry basis), 1.95% moisture, 8.56% fat, and 4.30% ash. It was light cream to pale yellow in appearance, bland in flavor, and dispersible in water. The isolate exhibited medium emulsification capacity and softer gel formation relative to the dual-purity CPI, making it suitable for non-dairy yogurts, cream-style spreads, and plantbased beverages.

[0200] CFP Product

[0201] The insoluble solids separated during alkaline extraction were dried as in Example 4, yielding a CFP with >25% protein (dry basis), 50-65% dietary fiber, <8% moisture.VHPM 00773. OOlWOl Example 6: Preparation of Functional Cluster Bean Protein Concentrate (FCPC) by pH Adjustment and Heat Treatment

[0202] Feed material

[0203] Cluster bean protein concentrate (90% protein dry basis, 8-9% moisture, <1% fat, and -5% ash) was prepared by the dual -purity ethanol extraction process described in Example 1. This CPC was cream -colored, bland in flavor, and readily dispersible in water, but showed limited ability to form stable gels or emulsions in functional tests.

[0204] pH adjustment and heat treatment

[0205] The CPC was dispersed in water at a solids-to-water ratio of 1 : 10 (w / w) and adjusted to pH 7.5 using sodium hydroxide solution. The slurry was then subjected to direct steam injection at 135 °C for 5-10 seconds in a holding tube, followed by rapid vacuum cooling to -55 °C.

[0206] Spray drying

[0207] The neutralized slurry was spray dried in a tail-form dryer equipped with a high-pressure nozzle (operated at -4000 psi; inlet 187 °C, outlet 92 °C), yielding a fine powder with a final moisture level of <6%. The resulting product was designated FCPC.

[0208] Functional evaluation

[0209] The untreated CPC and the FCPC were tested in the 1 :4 Gel and 1 :4:4 Emulsion systems (see Functional Characterization Methods).

[0210] - 1:4 Gel: The untreated CPC formed only a weak gel with water release, whereas the FCPC formed a firm, cohesive gel with visible elasticity, reduced syneresis, and maintained structural integrity when split with a knife.

[0211] - 1 :4:4 Emulsion: The untreated CPC emulsions were weak and prone to phase separation, whereas the FCPC produced stable emulsions with good body, uniform texture, and minimal oiling-off after cooling.

[0212] Product

[0213] The FCPC contained >84% protein (dry basis), with moisture <6% and ash -5%.

[0214] Compared to untreated CPC, the FCPC exhibited significantly enhanced gelation and emulsification properties, making it suitable for applications requiring structure formation, such as plant-based meat analogues, processed cheese, or high-protein spreads.

[0215] Visual documentation of the functionality comparison is provided in Figure 6, where the upper panel shows the untreated CPC and the lower panel shows the FCPC. The top images correspond to the 1 :4 Gel test and the bottom images correspond to the 1 :4:4 Emulsion test, both prepared and evaluated according to the Functional Characterization Methods.

[0216] Example 7: Preparation of Texturized Cluster Bean Protein Concentrates (TVP Form)VHPM 00773. OOlWOl Cluster bean protein concentrates (CPC) were mixed in equal proportions (~11 kg each) and milled together in a Prater Sterling Rotomill 1300. Analytical results from the two lots indicated protein levels of -84-85% (dry basis), -4-5% ash, and -3-4% fat. The combined mixed lot was estimated at -84.5% protein (dry basis), -4.5% ash, and -3.5% fat, with -7-8% moisture.

[0217] Extrusion conditions

[0218] Extrusion was performed on a lab-scale co-rotating twin-screw extruder (MPF-19, APV Baker Ltd., Peterborough, UK). The feed material was hydrated to 27% feed moisture (wet basis). The extrusion barrel temperature profile was set at 85-105-125-130-135 °C, with screw speed of 250 rpm and feed rate of 2.62 kg / h (dry basis). Extrudates were cut to size upon exit, dried in a hot-air oven at 40 °C overnight, and packaged.

[0219] Product

[0220] The resulting texturized CPC (TVP form) had 84.5% protein (dry basis), 4% ash, and -3.5% fat and formed expanded, porous, fibrous chunks typical of soy or pea texturized vegetable proteins. Internal structure (examined by knife-sectioning under microscope) showed well-developed fibrous alignment, high aeration, and high water-holding capacity. The material rehydrated rapidly and maintained spongy, fibrous texture, confirming suitability for meat analogue and savory applications.

[0221] Example 8: Preparation of Cluster Bean Protein-Tapioca Starch Extruded High-Protein Crisps (Figure 5)

[0222] Feed material

[0223] The same CPC80 mixed lot as described in Example 7 (84.5% protein dry basis) was blended with food-grade tapioca starch at a ratio of 70:30 (w / w). Based on this formulation, the calculated protein content of the dry blend was 59% protein (dry basis), with 3% fat and 4% ash.

[0224] Extrusion conditions

[0225] Extrusion was performed on the same lab-scale co-rotating twin-screw extruder (MPF-19, APV Baker Ltd., Peterborough, UK). The feed mixture was adjusted to 25% feed moisture (dry basis). The barrel temperature profile was 75-95-115-130-135 °C, with screw speed of 250 rpm and feed rate of 1.62 kg / h. Extrudates were cut to size upon exit, dried in a hot-air oven at 40 °C overnight, and packaged.

[0226] Product

[0227] The resulting extruded crisps (CPC-Tapioca 70:30) were light, airy, and expanded with uniform pore distribution. Analytical composition of the final product was estimated at 59% protein (dry basis), with <3% fat, 4% ash, and 7% moisture after drying. The crisps exhibited aVHPM 00773. OOlWOl crunchy, puffed texture characteristic of high-protein extruded snacks, comparable to soy-starch crisps.

[0228] Example 9: Preparation of a Ready-to-Drink (RTD) Protein Beverage Using Cluster Bean Protein Isolate (CPI)

[0229] A high-protein, ready -to-drink (RTD) beverage was formulated using cluster bean protein isolate (CPI) prepared according to the method of Example 4. The CPI used contained >90% protein (dry basis), <3.5% fat, 3-6% ash, and moisture <6%. The CPI exhibited a bland flavor, light color, and excellent dispersibility in water.

[0230] Table 1 — RTD Beverage Formula

[0231] Ingredient % w / w

[0232] Water 85.50

[0233] Cluster Bean Protein Isolate (CPI) 10.00

[0234] Tagatose 2.50

[0235] Sucrose 1.50

[0236] Sunflower oil 0.50

[0237] Lecithin (emulsifier) 0.30

[0238] Vitamins & Minerals premix 0.20

[0239] Natural flavors 0.50

[0240] The CPI was dispersed in water at 55-60 °C under moderate agitation using a Silverson L5M-high shear rotor / stator laboratory mixer at 5,000 rpm until fully hydrated. Tagatose and sucrose were dissolved separately in warm water (60 °C) and added to the CPI slurry. Sunflower oil and lecithin were pre-blended and emulsified into the protein base with speed increased to 10,000 rpm for high-shear homogenization. Vitamins, minerals, and natural flavors were added last. The beverage was then homogenized in a two stage GEA Panther Lab Homogenizer 3006 (200 / 50 bar), pasteurized at 85 °C for 2 minutes in an induction batch pasteurizer, cooled to 4 °C, and aseptically filled into bottles.

[0241] The final RTD beverage had 10 g protein / 100 ml, a smooth, low-grit mouthfeel, bland taste, and stable emulsion with no visible sediment after 14 days refrigerated storage.

[0242] Example 10: Preparation of a Dry-Blend Instant Protein Beverage Using CPC.

[0243] A dry instant beverage powder was produced using CPC prepared as described in Example 2 of this invention. The CPC contained >80% protein (dry basis), <3.5% fat, 3-6% ash, moisture <8%, and was fine-milled (<150 pm ) to improve dispersibility and reduce grittiness using aVHPM 00773. OOlWOl Prater- Sterling Rotomill 1300 connected to a Prater- Sterling MAC-0 Air classifier. The Rotomill 1300 was set to Mill Wheel Speed of 6960 rpm and a power level of 60 Hz. The MAC-0 Air Classifier was set to a power level of 5 Hz.

[0244] The fine-milled CPC was formulated according to Table 2:

[0245] Table 2 — Instant Beverage Powder Formula

[0246] Ingredient % w / w

[0247] CPC (<150 pm ) 47.15%

[0248] Sugar 36.04%

[0249] Gum Acacia 1.36%

[0250] Guar / Xanthan Blend 0.56%

[0251] Commercial Creamer Blend 10.48%

[0252] Natural Vanilla Flavors 4.42%

[0253] Total 100.00

[0254] A 56.5 g serving of the blended powder was mixed with 10 oz (-295 ml) of water, corresponding to approximately 20 g of protein per serving. The powder showed excellent dispersibility, achieving full solubilization within 30 seconds using a standard shaker bottle at 25 °C.

[0255] The resulting beverage was smooth and homogeneous, with no visible sedimentation or foam, a neutral, non-beany flavor, and a light cream suitable for flavored ready -to-drink or instant applications.

[0256] Example 11: Preparation of Gluten-Free Bread Fortified With CPC

[0257] Gluten-free bread was prepared by partially replacing starch-based flour with CPC to achieve a high-protein bread. The CPC contained >80% protein (dry basis), <3.5% fat, 3-6% ash, moisture <8%, and was fine-milled to <150 pm as described in example 10 to improve incorporation and mouthfeel.

[0258] Table 3 — Gluten-Free High-Protein Bread Formula

[0259] Ingredient % w / w

[0260] Rice flour 40.00

[0261] Fine-milled Dual-Purity CPC 25.00

[0262] Tapioca starch 15.00Ingredient % w / w

[0263] Sunflower oil 6.00

[0264] Sucrose-Maple mix (50 / 50) 6.00

[0265] Whole egg powder 2.00

[0266] Salt 1.50

[0267] Hydroxypropyl methylcellulose (HPMC) 1.00

[0268] Yeast 1.00

[0269] Water 1.50 (adjusted for dough hydration)

[0270] Total 100.00

[0271] Dry ingredients were blended, oil and water were added to form dough, and yeast was incorporated. The dough was proofed for 60 min at 35 °C and baked at 180 °C for 35 min. The final bread showed 14-16 g protein / 100 g, good loaf volume, light cream color, and bland taste. The CPC incorporation provided higher protein content while maintaining acceptable texture and sliceability compared to conventional gluten-free bread.

[0272] Example 12: Preparation of Plant-Based Yogurt Using Cluster Bean Protein Isolate (CPI) A spoonable, dairy-free yogurt alternative was formulated using cluster bean protein isolate (CPI) obtained according to Example 4.

[0273] The CPI contained >90% protein (dry basis), <3.5% fat, 3-6% ash, and moisture <6%. The CPI was bland in flavor and easily hydrated.

[0274] Table 4 — Plant-Based Yogurt Formula

[0275] Ingredient % w / w

[0276] Water 78.00

[0277] Cluster Bean Protein Isolate (CPI) 9.00

[0278] Sunflower oil 4.00

[0279] Sucrose 6.00

[0280] Tapioca starch 1.50

[0281] Pectin 0.60

[0282] Locust bean gum 0.40

[0283] Starter culture 0.10

[0284] Vitamins & minerals 0.30

[0285] N atural fl avor s 0.10VHPM 00773. OOlWOl The CPI was dispersed in water at 55-60 °C a Silverson L5M-high shear rotor / stator laboratory mixer at 2,000 rpm until fully hydrated. Starch and hydrocolloids were hydrated separately and blended into the CPI slurry. Oil was pre-emulsified with sucrose and added while mixing. The mixture was homogenized at 10,000 rpm using the Silverson L5M-followed by high pressure homogenization using a two stage GEA Panther Lab Homogenizer 3006 (200 / 50 bar). The homogenized slurry was then pasteurized at 90 °C for 5 minutes in an induction batch pasteurizer, cooled to 40 °C, and inoculated with a standard yogurt starter culture (Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus). Fermentation proceeded until pH 4.6, after which the yogurt was cooled to 4 °C and flavored.

[0286] The final product exhibited 8 g protein / 100 g, smooth creamy texture, and mild, dairy -like flavor with good emulsion and gel stability.

[0287] Example 13: Preparation of Meat Analogue Patty Using Textured CPC (TCPC)

[0288] A plant-based burger analogue was formulated using TCPC (>80% protein dry basis, <6% fat) produced from CPC as described in example 7.

[0289] Table 5 — Meat Analogue Patty Formula

[0290] Ingredient % w / w

[0291] Hydrated TCPC (at 30% moisture) 50.00

[0292] Wheat gluten 11.00

[0293] Soybean oil 11.00

[0294] Sucrose 2.50

[0295] Methylcellulose 1.50

[0296] Natural color / flavor 0.50

[0297] Salt 1.20

[0298] Seasoning blend 1.80

[0299] Water (adjusted to dough moisture -55%) 20.00

[0300] TCPC was hydrated separately in warm water (40 °C) for 30 min. Hydrated proteins were blended with wheat gluten, methylcellulose, and seasonings in a planetary mixer to form a cohesive dough. Oil was emulsified in, followed by sweeteners and flavors. Patties were formed and cooked to an internal temperature of 72 °C.

[0301] The resulting patties had >23 g protein / 100 g, fibrous meat-like texture, good bite, and bland baseline flavor suitable for further seasoning or marination.VHPM 00773. OOlWOl

[0302] Example 14: Preparation of Processed Meat Sausage With Functionalized and Texturized CPC (FCPC + TCPC)

[0303] A hybrid meat product (reduced meat content) was formulated using functionalized CPC (FCPC) and TCPC as clean-label extenders.

[0304] FCPC contained >80% protein (dry basis), <3.5% fat, improved gelation and emulsion stability produced as described in example 6..

[0305] • TCPC contained >80% protein (dry basis), <6% fat, porous fibrous structure produced as described in example 7.

[0306] Table 6 — Reduced-Meat Sausage Formula

[0307] Ingredient % w / w

[0308] Ground pork (30% fat) 50.00

[0309] Hydrated FCPC (20% solids slurry) 20.00

[0310] Hydrated TCPC (30% moisture) 10.00

[0311] Lard Fat 6.00

[0312] Icewater 10.00

[0313] Salt 1.80

[0314] Phosphates 0.50

[0315] Spices & flavors 1.50

[0316] Sugar 0.20

[0317] FCPC slurry was prepared by hydrating FCPC powder to 20% solids. TCPC was hydrated separately to 30% moisture. Meat was chopped in a bench top food processor with salt, phosphates, and ice water to form an emulsion base. FCPC slurry and hydrated TCPC were incorporated under moderate shear, followed by oil, sweeteners, and spices. The mixture was stuffed into casings using a manual sausage stuffer and cooked to 72 °C for 60 minutes.

[0318] The finished sausages were lighter in color, bland in taste, with high protein content (~18 g / 100 g final product) and excellent water / oil retention with no visible fat separation.

[0319] Example 15: Preparation of High-Protein Chewy Nutrition Bar Using CPC

[0320] A chewy protein bar was formulated using CPC produced according to from Example 2. The CPC contained >80% protein (dry basis), <3.5% fat, 3-6% ash, moisture <8%, and was fine-milled to <150 pm as described in example 10 to improve incorporation and mouthfeel.Table 7 — Chewy Protein Bar Formula

[0321] Ingredient % w / w

[0322] Glucose syrup 25.00

[0323] Sucrose / Honey mix (60 / 40) 20.00

[0324] Glycerin 5.00

[0325] Sunflower oil 4.00

[0326] Cluster Bean Protein Concentrate (CPC) 32.00

[0327] Soluble corn fiber 8.00

[0328] Cocoa powder 2.00

[0329] Lecithin 1.00

[0330] Salt 0.50

[0331] Natural flavors 0.50

[0332] Vitamins & minerals 2.00

[0333] The CPC powder was pre-blended with cocoa, salt, and lecithin to improve dispersion. Glucose syrup, tagatose, sucrose, glycerin, and oil were heated to 65-70 °C under gentle stirring to form a binder syrup. Dry ingredients were gradually added to the syrup and mixed until a cohesive dough was formed. The mass was sheeted to ~15 mm thickness, cooled to <30 °C, and cut into 60 g bars.

[0334] The resulting bar contained 20 g protein per 60 g serving, had a soft, chewy texture, and a clean, bland protein taste suitable for flavored coatings or inclusions.

[0335] Example 16: Preparation of High-Protein Crunchy Nutritional Bar Using High-Protein Crisps (HPC) Produced From CPC

[0336] High-protein crisps (HPC) contains 70% protein were produced by extrusion of CPC (>80% protein, <3.5% fat) blended with tapioca starch as described in Example 8, and then incorporated into a layered nutrition bar.

[0337] Table 8 — Crunchy Protein Bar Formula

[0338] Ingredient % w / w

[0339] High-Protein Crisps (HPC, 70% protein) 35.00

[0340] Glucose syrup 25.00

[0341] Sucrose 10.00Ingredient % w / w

[0342] Peanut butter 10.00

[0343] Sunflower oil 4.00

[0344] Glycerin 5.00

[0345] Soluble corn fiber 7.00

[0346] Natural flavors & salt 1.50

[0347] Vitamin / mineral premix 2.50

[0348] Binder syrup was prepared by heating glucose syrup, sucrose, glycerin, oil, and peanut butter to ~65 °C until uniform. Crisps, soluble fiber, and dry fortification blend were gently folded into the binder to avoid crushing the HPC structure. The mixture was manually pressed into bars and cooled to <25 °C. The final product was crispy and light, supplying ~22 g protein per 60 g bar, with excellent structural integrity and clean, neutral flavor.

[0349] Example 17: Preparation of High-Protein Low-Sugar Breakfast Cereal Produced From CPC

[0350] A high-protein, low-sugar breakfast cereal was produced using CPC produced according to from Example 2. The CPC contained >80% protein (dry basis), <3.5% fat, 3-6% ash, moisture <8%, and was fine-milled to <150 pm as described in example 10.

[0351] Table 10 — High-Protein Extruded Cereal Formula

[0352] % (w / w,

[0353] Ingredient

[0354] dry basis)

[0355] Cluster Bean Protein Concentrate (CPC) 38.0

[0356] Tapioca starch 24.0

[0357] Potato fiber 10.0

[0358] Com fiber 8.0

[0359] Inulin fiber 6.0

[0360] Guar gum 1.0

[0361] Tagatose + sucrose blend (1:1) 7.0

[0362] Baking soda 0.4

[0363] Monocalcium phosphate 0.3

[0364] Calcium carbonate 0.3VHPM 00773. OOlWOl % (w / w,

[0365] Ingredient

[0366] dry basis)

[0367] Yeast extract (fl avor) 0.5

[0368] Salt 0.5

[0369] Total 100.0

[0370] Processing

[0371] All dry ingredients were premixed in a ribbon blender for 5 min to achieve uniform distribution. Moisture was adjusted to 18-22 % using water pre-heated to 50 °C.

[0372] The hydrated blend was extruded on a twin-screw extruder (MPF-19, APV Baker Ltd., Peterborough, UK) under the following representative profile:

[0373] • Barrel zones 1-5 temperatures: 80 °C — 95 °C — 120 °C — 130 °C — 135 °C

[0374] • Screw speed: 250 rpm

[0375] • Feed rate: 1.8 kg / h (dry basis)

[0376] The extrudate was cut to 8-10 mm pillow shapes at the die and dried in a forced-air oven at 40 °C overnight to a final moisture of < 6 %.

[0377] Product Characteristics

[0378] The resulting cereal was light-brown, crisp, and crunchy, with 41 % protein (dry basis), 4 % fat, and 4.5 % ash. It displayed clean, bland flavor and excellent crunch retention in milk and yogurt systems.

[0379] Analytical Methods

[0380] The following analytical methods were used in the above examples:

[0381] 1. Protein Content

[0382] Determined by Kjeldahl nitrogen analysis (AO AC 981.10) or Dumas combustion (AO AC 992.15), using a nitrogen-to-protein conversion factor of 6.25.

[0383] 2. Moisture Content

[0384] Determined by oven drying at 105 °C to constant weight (AO AC 925.10).

[0385] 3. Ash Content

[0386] Determined by muffle furnace incineration at 550 °C (AO AC 923.03).

[0387] 4. Crude Fat Content

[0388] Crude fat determined by Soxhlet extraction with petroleum ether (AO AC 920.39) or by acid-hydrolysis fat (AOAC 922.06), depending on the testing laboratory.

[0389] 5. Gelation Capacity

[0390] A 1 :4 (w / w) protein-to-water slurry was prepared in a standard food processor, heated toVHPM 00773. OOlWOl 80 °C for 40 minutes, then chilled overnight at 4 °C. Gel firmness and homogeneity were visually inspected and manually tested for firmness / break strength.

[0391] 6. Emulsion Stability / Capacity

[0392] A 1:4:4 (protein:water:oil) emulsion was prepared in a standard food processor, heated to 80 °C for 40 minutes, then chilled overnight at 4 °C. Texture, phase separation, and stability were visually examined and rated.

[0393] 7. Color

[0394] Assessed by visual comparison of powders and rehydrated slurries against a white standard background under daylight-equivalent lighting to describe lightness and absence of dark or brown hues.

[0395] 8. Flavor and Odor

[0396] Flavor and odor were evaluated by a trained internal sensory panel (n=3) under controlled conditions. Panelists scored overall odor intensity, off-notes (beany, bitter, saponin), and flavor neutrality.

[0397] Although the foregoing specification and examples fully disclose and enable the present invention, they are not intended to limit the scope of the invention, which is defined by the claims appended hereto.

[0398] All publications, patents and patent applications are incorporated herein by reference. While in the foregoing specification this invention has been described in relation to certain embodiments thereof, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein may be varied considerably without departing from the basic principles of the invention.

[0399] The use of the terms “a” and “an” and “the” and similar referents in the context of describing the invention are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0400] As used herein, the term “about,” when referring to a value is meant to encompass variations of, in some embodiments ± 50%, in some embodiments ± 20%, in some embodiments ± 10%, in some embodiments ± 5%, in some embodiments ± 1%, in some embodiments ± 0.5%, and in some embodiments ± 0.1% from the specified amount, as such variations are appropriate to perform the disclosed methods or employ the disclosed compositions.

[0401] The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to”) unless otherwise noted.

[0402] Recitation of ranges of values herein are merely intended to serve as a shorthand method ofVHPM 00773. OOlWOl referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. It is understood that embodiments described herein include “consisting of’ and / or “consisting essentially of’ embodiments.

[0403] The transitional phrase “consisting of’ excludes any element, step, or ingredient not specified in the claim.

[0404] “Consisting essentially of’ generally limits a feature, compound, composition or method to the recited elements and / or steps but does not exclude the possibility of additional elements and / or steps that do not materially affect the function, compound, composition and / or characteristics of the recited feature, compound, composition or method. The transitional phrase “consisting essentially of’ limits the scope of a claim to the specified materials or steps and those that do not materially affect the basic and novel character! stic(s) of the claimed invention.

[0405] All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0406] Embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein.

[0407] Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

What is claimed is:

1. A method of producing a cluster bean protein concentrate (CPC), comprising:(a) providing un-toasted cluster bean germ;(b) contacting the germ with a first solvent comprising ethanol of at least 190 proof to separate out a first protein-rich solid by removing at least one of;(c) contacting the first protein-rich solid with a second solvent comprising aqueous ethanol of about 120-160 proof to separate out a second protein-rich solid; and(d) desolventizing and drying the second protein-rich solid under vacuum conditions at a temperature below about 75 °C to produce the CPC.

2. The method of claim 1, wherein contacting the germ with the first solvent removes at least one of a non-polar lipid, a pigment, or a resinous impurity; and wherein contacting the first protein-rich solid with the second solvent comprises removing at least one of a saponin, a sugar, or a polar impurity.

3. The method of claim 1 or 2, wherein the contacting steps are carried out in a Nutsche-type filter-dryer or a continuous counter-current extractor at about 40°C -60C with solvent-to-solid ratios of about 10:1 to about 2:1 (w / w).

4. The method of any of claims 1-3, further comprising sieving and color sorting the germ prior to the contacting steps.

5. The method of any of claims 1-4, wherein the germ is milled, pelletized or flaked at a temperature below about 75 °C prior to the contacting steps.

6. The method of any of claims 1-5, wherein the ethanol or the aqueous ethanol is a specially denatured alcohol (SDA) comprising methanol, isopropanol, n-propanol, or another C1-C3 alkanol.

7. The CPC produced by the method of any of claims 1-6, the CPC comprising at least about 80% protein on a dry basis and less than about 3.5% fat.

8. The CPC of claim 7, wherein the CPC comprises greater than about 84% protein on a dry basis and less than about 1% fat.

9. A method of producing a cluster bean protein concentrate (CPC), comprising:(a) providing un-toasted cluster bean germ;(b) contacting the germ with an aqueous ethanol solvent of about 120-160 proof to produce a protein-rich solid; and(c) desolventizing and drying the protein-rich solid under vacuum conditions at a temperature below about 75 °C.

10. The method of claim 9, wherein the germ is pre-treated by sieving, color sorting, milling, pelletizing, or flaking.

11. The method of claim 9 or 10, wherein the aqueous ethanol solvent is a or 10, wherein the contacting steps are carried out in a Nutsche-type filter-dryer or a continuous counter-current extractor at about 40°C -60°C with solvent-to-solid ratios of about 10:1 to about 2:1 (w / w).

12. The method of claim 9 or 10, wherein the aqueous ethanol solvent is a specially denatured alcohol (SDA) comprising methanol, isopropanol, n-propanol, or another C1-C3 alkanol.

13. The CPC produced by the method of any of claims 9-11, the CPC comprising at least about 80 % protein on a dry basis and less than about 6 % fat.

14. A method of producing a cluster bean protein isolate (CPI), comprising:(a) hydrating the CPC produced by the method of any of claims 1-6 or 9-12 at a CPC raw material solids-to-water ratio of about 1 :7-l : 14 to produce a first slurry;(b) adjusting the first slurry to a pH of about 7.5-9.5 with a food-grade alkaline agent and applying high-shear mixing at 40-65 °C to produce solubilized proteins;(c) separating the solubilized proteins from insoluble fiber to produce a protein-rich liquid;(d) acidifying the protein-rich liquid to pH 4.3-4.8 to precipitate a protein solid;(e) resuspending and neutralizing the protein solid to a pH of about 6.0-8.0 to produce a second slurry;(f) subjecting the second slurry to direct steam injection at about 120-145 °C for about 5-30 seconds, followed by rapid cooling below about 70 °C, and drying to yield the CPI.

15. The method of claim 14, further comprising applying a functionalization treatment to the second slurry after step (e), wherein the functionalization treatment comprises at least one of an enzyme, a crosslinking agent, or high-pressure homogenization.

16. The method of claim 14 or 15, further comprising step (g) treating the dried CPI with coating or agglomeration.

17. The CPI produced by the method of any of claims 14-16, wherein the CPI comprises greater than about 94% protein on a dry basis and less than about 0.5% fat.

18. The CPI produced by the method of any of claims 14-16, wherein the CPI comprises greater than about 87% protein on a dry basis and less than about 8.5% fat.

19. A cluster bean fiber-protein fraction (CFP) comprising the insoluble fiber obtained in step (c) of claim 14, wherein the CFP comprises greater than about 25% protein on a dry basis, about 50-60% dietary fiber, and less than about 8% moisture.

20. A method of producing a functionalized CPC (FCPC), the method comprising hydrating the CPC produced by the method of any of claims 1-6 or 9-12 to produce a slurry, adjusting the pH to about 6-8, subjecting the slurry to direct steam injection at about 120-145 °C for about 5-30 seconds followed by rapid cooling below about 70 °C, and drying the slurry to yield the FCPC.

21. The FCPC produced by the method of claim 20, wherein the FCPC comprises at least 84 % protein on a dry basis, and less than 6 % moisture.

22. A method of producing a texturized CPC (TCPC), comprising hydrating the CPC produced by the method of any of claims 1-6 or 9-12 to 10-40% moisture, extruding at 80-150 °C using a single- or twin-screw food extruder to form a fibrous expanded product, and drying to less than 10 % moisture.

23. The TCPC produced by the method of claim 22, wherein the TCPC comprises at least 80 % protein on a dry basis, less than 6 % fat, less than 10 % moisture.

24. A method of producing a texturized CFP (TCFP), comprising hydrating the insoluble fiber obtained in step (c) of claim 14 to 10-40% moisture, extruding at 80-150 °C using a single- or twin-screw food extruder to form a fibrous expanded product, and drying to less than 10 % moisture.

25. The TCFP produced by the method of claim 24, wherein the TCFP comprises at least 25 % protein on a dry basis, and less than 10 % moisture.

26. A method of producing high-protein extruded crisps (HPC), the method comprising blending 50-90 % CPC produced from the method of any of claims 1-6 or 9-12 with 10-50% starch, adjusting moisture to 10-20%, extruding in a single- or twin-screw food extruder at 70-150 °C, and drying to 5-8% moisture to yield expanded crisps.

27. The HPC produced by the method of claim 26, wherein the HPC comprises greater than 50% protein on a dry basis, less than 4% fat, and less than 10% moisture.

28. A food product comprising the CPC of claims 7, 8, or 13.

29. The food product of claim 28, wherein the food product is selected from the group consisting of a high-protein beverage, a dry beverage mix, a baked product, a gluten-free bakery item, a high-protein nutrition bar, a chewy or crunchy bar, a breakfast cereal, a high-protein extruded snack, a meat analog, a hybrid meat or processed meat product, a pasta, a soup, a sauce, a plant-based dairy alternative, a non-soy plant milk, a liquid or powdered dietetic formula, a health or sports nutrition supplement, a meal replacement, a confectionery product, or a functional food ingredient.

30. A food product comprising the CPI of claim 17 or 18.

31. The food product of claim 30, wherein the food product is selected from the group consisting of a ready-to-drink (RTD) protein beverage, an instant beverage powder, a non-dairy yogurt, a dairy analog product, a non-soy plant milk, a high-protein smoothie, a frozen dessert, a protein shake, a meal replacement beverage, a dietetic or clinical nutrition formula, a sports nutrition drink, or a functional beverage.

32. A food product comprising the CFP of claim 19.

33. The food product of claim 32, wherein the food product is selected from the group consisting of a fiber-enriched baked product, a gluten-free bread or cracker, a high-fiber cereal, a fiber-enriched snack, a nutrition bar, a meat analog or extender, a pasta, a plant-based dough, a high-fiber beverage, a smoothie powder, a functional prebiotic food, or a dietary supplement.

34. A food product comprising the FCPC of claim 21.

35. The food product of claim 34, wherein the food product is selected from the group consisting of a meat analog, a processed meat product, a sausage, a hybrid meat formulation, a plant-based cheese, a spread or dip, an emulsified dressing, a high-protein dessert, a smoothie powder, or a functional binder in food formulations.

36. A food product comprising the TCPC of claim 23, wherein the TCPC exhibits a fibrous, meat-like texture.

37. The food product of claim 36, wherein the food product is selected from the group consisting of a plant-based burger, a meat analog patty, a textured vegetable protein (TVP) replacement, a plant-based sausage, a meat hybrid, a crumb or chunk analog, a high-moisture extruded product, a Pizza topping, a ready -to-cook plant protein base, or a refrigerated or frozen meal.

38. A food product comprising the HPC of claim 27.

39. The food product of claim 38, wherein the food product is selected from the group consisting of a ready-to-eat protein snack, a protein-enriched breakfast cereal, a high-protein bar inclusion, a baked granola, a trail mix, a functional food topping, or a crisp inclusion in nutrition bars or dairy analogs.

40. A food product comprising the TCFP composition of claim 25, wherein the TCFP comprises at least about 40% dietary fiber.

41. The food product of claim 40, wherein the food product is selected from the group consisting of a meat analog, a plant-based ground product, a crumb or filler for meat hybrid, asnack extrusion, a pasta, a baked savory snack, a pizza topping, a meat extender, or a high-fiber textured ingredient.

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