Legume protein product and process of preparing the same

Abstract

The present disclosure relates to a legume protein product and a process for preparing the same. The process comprises the steps of providing a legume protein containing material; purifying the legume protein containing material to obtain a purified material; treating the purified material with one or more enzymes, followed with one or more encapsulating agents, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and removing the encapsulated flavor compounds from the mixture of treated components to obtain the legume protein product. The legume protein product has a reduced flavor compound content and / or an increased protein content compared to an equivalent untreated legume protein product.

Description

LEGUME PROTEIN PRODUCT AND PROCESS OF PREPARING THE SAMECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of European Application No. 24176070.1, filed May 15, 2024, which is hereby incorporated by reference in its entirety.FIELD OF INVENTION

[0002] This invention relates to the field of plant-based protein products, in particular legume protein products.BACKGROUND

[0003] Soybean (Glycine max) is a rich legume source of proteins exhibiting a high nutritional value, in addition to bioactive compounds with potential health benefits. Although the demand for plant-based protein foods has been increasing considerably in the past years, improving sensory aspects, in particular flavor, is still a major challenge for the food industry.

[0004] With the target of producing palatable plant-based protein foods with better quality, there exists strong interests to apply advanced interventions to improve the nutritional, sensory, and functional properties of soy-based products, in particular soy flour.

[0005] Typically, soy flour is classified as a food grade enzyme active flour prepared from clean, whole soybeans, which have been dehulled, solvent extracted, desolventized, and ground, having a protein content of less than 50 wt% (dry basis). It can also be referred and labeled as “defatted soy flour”.

[0006] To overcome the unpleasant sensory defects, commercial soy -based products can be supplemented with artificial colorants, flavor enhancers, and other molecules. However, considering the presence of artificial ingredients in the products, soy products mixed with natural plant and / or animal-based ingredients, or soy products without any of such ingredients, are becoming the latest trend.

[0007] Products made from soy protein tend to have off-notes, which are usually described by consumers as “beany”, "grassy", "leafy", "malty", "fatty", "fishy", among other aroma attributes.

[0008] The off-notes and their precursors (e.g., flavor precursors) can exist as protein-bound complexes present in soy flour and can be attributed to the volatile compounds (e.g., short-chain aliphatic alcohols, ketones, and aldehydes).

[0009] More specifically, residual phospholipids and free fatty acids in soy flour are oxidized and decomposed by endogenous lipoxygenase. During soybean oil extraction along with soy flourproduction (as subproduct), soybeans are subjected to several stages such as soaking, squeezing, and grinding, which cause the exposure of lipoxygenase to soy lipids leading to deterioration.

[0010] The volatile compounds are mainly divided into fatty aldehydes, fatty alcohols, fatty ketones, furans, furan derivatives, and aromatic compounds. Hexanal is one of the most important volatile compounds with the highest threshold. However, non-volatile compounds, including flavonoids, tannins, saponins, phenolic acids and phenolic compounds, and certain amino acids, can promote the formation of unpleasant odorants.

[0011] Therefore, there is a need for a plant protein product (e.g., soy protein product) having off notes reduced or completely removed, and an improved process for preparing such plant protein product.SUMMARY

[0012] The present disclosure provides a process for preparing a legume protein product comprising the steps of: (a), providing a legume protein containing material; (b). purifying the legume protein containing material to obtain a purified material; (c). treating the purified material with one or more enzymes, followed with one or more encapsulating agents, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and (d). removing the encapsulated flavor compounds from the mixture of treated components to obtain the legume protein product. The legume protein containing material is optionally treated with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes. The legume protein product has a reduced flavor compound content and / or an increased protein content compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (d).

[0013] The present disclosure provides a process for reducing flavor compound content of a legume protein product, comprising the steps of: (a), purifying a legume protein containing material, optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; (b). treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; (c). removing the encapsulated flavor compounds to obtain the legume protein product. The legume protein product has a flavor compound content reduced by at least 50%, preferably at least 90%, more preferably at least 98%, even more preferably 100%, compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).

[0014] The present disclosure provides a process for increasing protein content of a legume protein product, comprising the steps of: (a), purifying a legume protein containing material,optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; (b). treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; (c). removing the encapsulated flavor compounds to obtain the legume protein product. The legume protein product has an increased protein content by at least 10% compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).

[0015] The present disclosure provides a legume protein product having a reduced flavor compound content and / or an increased protein content compared to an equivalent untreated legume protein product that has not been subject to any process of the present disclosure.

[0016] The present disclosure provides a food product comprising the legume protein product of the present disclosure.

[0017] The present disclosure also provides a use of the legume protein product of the present disclosure in preparing a food product.

[0018] The present disclosure also covers the following aspects:Clause 1. A process for preparing a legume protein product, comprising the steps of: a. providing a legume protein containing material; b. purifying the legume protein containing material to obtain a purified material; c. treating the purified material with one or more enzymes, followed with one or more encapsulating agents, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and d. removing the encapsulated flavor compounds from the mixture of treated components to obtain the legume protein product; wherein the legume protein containing material is optionally treated with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes; and wherein the legume protein product has a reduced flavor compound content and / or an increased protein content compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (d).Clause 2. The process of clause 1, wherein the enzyme and encapsulating agent are not simultaneously added to the purified material.Clause 3. The process of any of the preceding clauses, wherein the legume protein containing material comprises legume protein selected from the group consisting of soy protein, fava protein, pea protein, chickpea protein, lupin protein, beans protein, mung bean protein, lentil protein, and any combinations thereof.Clause 4. The process of any of the preceding clauses, wherein the enzyme is lipase.Clause 5. The process of any of the preceding clauses, wherein the enzyme is a phospholipase selected from the group consisting of phospholipase A2 (PLA2), phospholipase Al (PLA1), phospholipase C (PLC), phospholipase D (PLD), and any combinations thereof.Clause 6. The process of any of the preceding clauses, wherein the encapsulating agent is selected from the group consisting of cyclodextrin, starch, starch derivatives, cellulose, cellulose derivatives, plant exudates, plant extracts, marine extracts, microbial polysaccharides, animal polysaccharides, and any combinations thereof.Clause 7. The process of any of the preceding clauses, wherein the cyclodextrin is selected from the group consisting of alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, and any combinations thereof.Clause 8. The process of any of the preceding clauses, wherein the flavor compounds are selected from the group consisting of phospholipid, phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.Clause 9. The process of any of the preceding clauses, wherein the purifying step (b) further comprising the steps of: i. mixing the legume protein containing material with an alkaline at a pH from 7 to 9 to obtain a protein-water mixture; ii. treating the protein-water mixture with ultrasound for 20 to 40 minutes at 2,500 W to obtain an ultrasonic-treated mixture; iii. incubating the ultrasonic-treated mixture at an incubation temperature from 35 to 50°C for an incubation period from 0.5 to 2 hours to obtain an incubated material; and iv. precipitating the incubated material by an acid to a pH from 4 to 6 to obtain the purified material.Clause 10. The process of any of the preceding clauses, wherein the one or more enzymes is phospholipase and the one or more encapsulating agents is cyclodextrin.Clause 11. A process for reducing flavor compound content of a legume protein product, comprising the steps of: a. purifying a legume protein containing material, optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; b. treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and c. removing the encapsulated flavor compounds to obtain the legume protein product; wherein the legume protein product has a flavor compound content reduced by at least50%, preferably at least 90%, more preferably at least 98%, even more preferably 100%,compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).Clause 12. The process of clause 11, wherein the flavor compounds are selected from the group consisting of phospholipid, phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.Clause 13. The process of any of clauses 11 to 12, wherein the flavor compounds are selected from the group consisting of phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.Clause 14. The process of any of claims 11 to 13, wherein the legume protein product has an aldehyde content, a ketone content, an alcohol content, and / or a furan content reduced by at least 50% compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).Clause 15. A process for increasing protein content of a legume protein product, comprising the steps of: a. purifying a legume protein containing material, optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; b. treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and c. removing the encapsulated flavor compounds to obtain the legume protein product; wherein the legume protein product has a protein content increased by at least 10% compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).Clause 16. The process of any of clauses 11 to 15, wherein the legume protein containing material comprises legume protein selected from the group consisting of soy protein, fava protein, pea protein, chickpea protein, lupin protein, beans protein, mung bean protein, lentil protein, and any combinations thereof.Clause 17. The process of any of the clauses 11 to 16, wherein the one or more enzymes is phospholipase and the one or more encapsulating agents is cyclodextrin.Clause 18. A legume protein product prepared by the process of any of the preceding clauses or by a process comprising or consisting of the steps of: a. providing a legume protein containing material; b. purifying the legume protein containing material to obtain a purified material, optionally treating the legume protein containing material with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes;c. adding one or more enzymes, followed by one or more encapsulating agents, to the purified material to obtain a mixture of treated components; and d. removing encapsulated flavor compounds from the mixture of treated components to obtain the legume protein.Clause 19. A legume protein product having a reduced flavor compound content as compared to an equivalent untreated legume protein product.Clause 20. A legume protein product having an increased protein content as compared to an equivalent untreated legume protein product.Clause 21. The legume protein of any of clauses 18 to 20, wherein the equivalent untreated legume protein product has not been subject to the process of any of clauses 1 to 18.Clause 22. The legume protein product of any of clauses 18 to 21 comprising one or more encapsulating agents, wherein the one or more encapsulating agents are used to prepare the legume protein product.Clause 23. The legume protein product of any of clauses 18 to 22 being free of any encapsulating agents.Clause 24. The legume protein product of any of clauses 18 to 23, wherein the encapsulating agent is selected from the group consisting of cyclodextrin, starch, starch derivatives, cellulose, cellulose derivatives, plant exudates, plant extracts, marine extracts, microbial polysaccharides, animal polysaccharides, and any combinations thereof.Clause 25. The legume protein product of any of clauses 18 to 24, having content of one or more flavor compounds reduced by at least 50%, preferably at least 90%, more preferably at least 98%, even more preferably 100%, as compared to an equivalent untreated legume protein product.Clause 26. The legume protein product of any of clauses 18 to 25, wherein the flavor compounds are selected from the group consisting of phospholipid, phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.Clause 27. The legume protein product of any of clauses 18 to 26, wherein the flavor compounds are selected from the group consisting of phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.Clause 28. The legume protein product of any of clauses 18 to 27, which has an aldehyde content reduced by at least 50% as compared to an equivalent untreated legume protein product, wherein the aldehyde is selected from the group consisting of pentanal, hexanal, 2-hexenal, heptanal, 2-heptanal, benzaldehyde, nonanal, 2-nonenal, and any combinations thereof.Clause 29. The legume protein product of any of clauses 18 to 28, which has a ketone content reduced by at least 50% as compared to an equivalent untreated legume protein product,wherein the ketone is selected from the group consisting of 2-heptanone, l-octen-3-one, and any combinations thereof.Clause 30. The legume protein product of any of clauses 18 to 29, which has an alcohol content reduced by at least 50% as compared to an equivalent untreated legume protein product, wherein the alcohol is selected from the group consisting of l-penten-3-ol, 1-pentanol, 1-hexanol and / or l-octen-3-ol, and any combinations thereof.Clause 31. The legume protein product of any of clauses 18 to 30, which has a furan content reduced by at least 50% as compared to an equivalent untreated legume protein product, wherein the furan is 2-pentylfuran.Clause 32. The legume protein product of any of clauses 18 to 31, being free of one or more flavor compounds selected from a group consisting of l-penten-3-ol, pentanal, 1-pentanol, 2- hexenal, 1-hexanol, 2-heptanone, heptanal, 2-heptenal, l-octen-3-ol, 2-pentylfuran, l-octen-3- one, 2-nonenal, and any combinations thereof.Clause 33. The legume protein product of any of clauses 18 to 32, having a non-detectable amount of one or more flavor compounds selected from a group consisting of l-penten-3-ol, pentanal, 1-pentanol, 2-hexenal, 1-hexanol, 2-heptanone, heptanal, 2-heptenal, l-octen-3-ol, 2- pentylfuran, l-octen-3-one, 2-nonenal, and any combinations thereof.Clause 35. The legume protein product of any of clauses 18 to 34, having a protein content increased by at least 10% as compared to an equivalent untreated legume protein product. Clause 36. A food product comprising the legume protein product of any of clauses 18 to 35. Clause 37. The food product of clause 36, wherein the food product is selected from a group consisting of meat substitute products, dairy substitute products, bakery products, beverage products, and convenience food products.Clause 38. Use of the legume protein product of any of clauses 18 to 35 in preparing a food product.DETAILED DESCRIPTION

[0019] Reference will now be made in detail to certain aspects of the disclosed subject matter. While the disclosed subject matter will be described in conjunction with the enumerated claims, it will be understood that the exemplified subject matter is not intended to limit the claims to the disclosed subject matter.

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this invention belongs. As used herein, each of the following terms has the meaning associated with it as defined below.

[0021] Unless expressly stated, ppm (parts per million), percentage, and ratios are based on a dry weight basis. Percentage based on a dry weight basis is also referred to as wt% below.

[0022] The term "for example," "for instance," "such as," or "including" as used herein is meant to introduce examples that further clarify more general subject matter. Unless otherwise specified, these examples are provided only as an aid for understanding the applications illustrated in the present disclosure and are not meant to be limiting in any fashion.

[0023] As used herein, “room temperature” or “RT” refers to a temperature between 20°C to 25°C.

[0024] In the processes described herein, the acts can be carried out in any order without departing from the principles of the disclosure, except when a temporal or operational sequence is explicitly recited. Furthermore, specified acts can be carried out concurrently unless explicit claim language recites that they be carried out separately. For example, a claimed act of doing X and a claimed act of doing Y can be conducted simultaneously within a single operation, and the resulting process will fall within the literal scope of the claimed process.

[0025] Described herein is a legume protein product and a process for preparing the legume protein product. The legume protein product has less flavor intensity and / or an increased protein content and is suitable for use as a protein source for incorporation into foods for human and / or animal consumption.Process for preparing a legume protein product

[0026] The present disclosure provides a process for preparing a legume protein product. The process comprises the steps of purifying a legume protein containing material; purifying the legume protein containing material to obtain a purified material; treating the purified material with one or more enzymes, followed with one or more encapsulating agents, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and removing the encapsulated flavor compounds from the mixture of treated components to obtain the legume protein product. The resulting legume protein product prepared by the process has one or more improved attributes compared to an equivalent untreated legume protein product; preferably, the one or more improved attributes may include, but may not be limited to, a reduced flavor compound content, an increased protein content, or any combinations thereof.

[0027] As described herein, an “equivalent untreated legume protein product” refers to an equivalent legume protein product that is obtained from conventional process technology, which may involve steps including soaking, separation, drying, grinding, or any combinations thereof, and that the equivalent legume protein product has not been subjected to any process of the instant invention as described in the present disclosure. An “equivalent legume protein product” refers toa legume protein product prepared from the same starting material used in the instant invention (e.g., a soy protein concentrate from the same batch).

[0028] The legume protein containing material serves as a starting material to the process and can include legume protein that may include, but may not be limited to, soy protein, fava protein, pea protein, chickpea protein, lupin protein, beans protein, mung bean protein, lentil protein, and any combinations thereof.

[0029] Preferably, the legume protein containing material can include legume protein with different concentrations (e.g., pulse, flour, concentrate, isolate, slurry, cake, low-protein stream, high-protein stream, or any intermediate products of the process as described the present disclosure) obtained from different botanical sources that may include, but may not be limited to, soy, fava, pea, chickpea, lupin, beans, mung bean, lentil, or any combinations thereof. More preferably, soy protein concentrate can be used as the starting material.

[0030] In one aspect, the legume protein containing material can have a protein content (initial protein content) in a range from 18 to 65 wt%, preferably from 40 to 60 wt%, more preferably from 45 to 55 wt%, on a dry basis.

[0031] The process can start with purifying (or pre-treating) the legume protein containing material in which surface-bound compounds, majority of which are water soluble compounds (e.g., phospholipids, off-note precursors such as phenolics, flavonoids), are removed from the legume protein containing material. Most of these removed compounds are responsible for carrying flavors (e.g., off-notes). Since water soluble carbohydrates, among other water-soluble compounds, can be washed out, protein content of the resulting purified material may be increased.

[0032] In one aspect, during the purifying step, the legume protein containing material is mixed with an alkaline solution at a pH from 7 to 9, from 7.5 to 9, from 7 to 8.5, or from 7 to 8, to obtain a protein-water mixture. The protein-water mixture is then incubated, preferably with stirring, at an incubating temperature from 35 to 50°C, from 40 to 50°C, or from 35 to 45°C, for an incubation period from 0.5 to 2 hours, from 0.8 to 2 hours, or from 0.5 to 1.5 hours to obtain an incubated material. The purified material is recovered by isoelectric precipitation.

[0033] The pH values of the isoelectric precipitation may vary dependent on the properties of the starting material. For example, when the starting material is a pea protein containing material, a soy protein containing material, a lentil protein containing material, a chickpea protein containing material, or a faba protein containing material, the corresponding pH for isoelectric precipitation may range from 3 to 7, from 4 to 7, from 4 to 6, from 4.5 to 6, from 4 to 5.5, or from 4 to 5.

[0034] Preferably, the purifying step can include an ultrasonic treatment to increase the efficiency of removing surface-bound compounds from the legume protein containing material. In such case, the protein-water mixture, obtained from mixing the legume protein containing material withwater at a pH from 7 to 9, from 7.5 to 9, from 7 to 8.5, or from 7 to 8, is treated with ultrasonic for an ultrasonic-treatment period from 20 to 40 minutes, from 25 to 40 minutes, or from 30 to 40 minutes to obtain an ultrasonic-treated mixture. The ultrasonic-treated mixture is then incubated, preferably with stirring, at an incubating temperature from 35 to 50°C, from 40 to 50°C, or from 35 to 45°C, for an incubation period from 0.5 to 2 hours, from 0.8 to 2 hours, or from 0.5 to 1.5 hours to obtain an incubated material. The incubated material undergoes isoelectric precipitation by acid to a pH from 4 to 6, from 4.5 to 6, from 4 to 5.5, or from 4 to 5, to recover the purified material.

[0035] The purified material is contacted with one or more enzymes to further remove compounds, most of which are responsible for carrying flavors (e.g., off-notes), from the purified material; preferably, flavor compounds are removed from the purified materials by the enzyme treatment.

[0036] An enzyme-treated material, which is free of, preferably completely free of, flavor compounds is obtained. One or more encapsulating agents are then added to the enzyme-treated material in which the free flavor compounds can then be trapped / encapsulated by the encapsulating agents.

[0037] Preferably, the enzymes are added to the purified material, followed by the addition of the encapsulating agents. More preferably, the enzymes and the encapsulating agents are not simultaneously added to the purified material.

[0038] Preferably, the enzyme may be lipase; more preferably, the lipase may include phospholipase superfamily; even more preferably, the phospholipase may include, but may not be limited to, phospholipase A2 (PLA2), phospholipase Al (PLA1), phospholipase C (PLC), phospholipase D (PLD), or any combinations thereof.

[0039] Preferably, the encapsulating agent may include, but may not be limited to, typical food grade wall material; more preferably food grade wall materials may include, but may not be limited to, cyclodextrin, starch and starch derivatives (e.g., maltodextrin, dextrins, poly dextrose), cellulose and cellulose derivatives (e.g., methylcellulose, citrus fiber), plant exudates and extracts (e.g., gum tragacanth, guar, locust bean, and gum arabic), marine extracts (e.g., carrageenan, alginates), microbial and animal polysaccharides (e.g., xanthan, gellan, curdlan, dextran, chitosan). More preferably, the cyclodextrin may include, but may not be limited to, alphacyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, or any combinations thereof; even more preferably, the cyclodextrin used in the process can be beta-cyclodextrin.

[0040] Upon completion of the reaction of the enzymes and the encapsulating agents, a mixture of treated components is obtained, which may include, but may not be limited to, encapsulated flavor compounds, unused encapsulating agents, and the legume protein product. Preferably, themixture of treated components may include encapsulated flavor compounds and the legume protein product. Then, encapsulated flavor compounds are removed to obtain the legume protein product; preferably, any unused encapsulating agents, if present, are removed together with the encapsulated flavor compounds.

[0041] In one aspect, unused encapsulating agents may be present in the legume protein product; preferably, the legume protein product may be free of any unused encapsulating agents.Process for reducing flavor compound content of a legume protein product

[0042] The present disclosure provides a process for reducing flavor compound content of a legume protein product. Preferably, the process may reduce or completely remove off-notes from the legume protein product. The process may also remove flavor precursors from the legume protein product before they develop into off-notes.

[0043] As described herein, an “off-note” or “off-taste” is an undesirable and / or unwanted flavor (e.g., taste, odor) and / or present in food products. An “off-note” may be originated from starting materials, derived from chemical changes, and / or developed by flavor precursors during food processing. A flavor precursor may not impart any off-notes when it is first added to a food product but may develop into one or more off-notes through reactions (e.g., oxidation, degradation) food processing. Unsaturated fatty acids (e.g., linoleic acid, linolenic acid) may be an example of a flavor precursor. Unsaturated fatty acids can be oxidized (by enzymes such as lipoxygenases) to form hydroperoxides, which are subsequently degraded into aldehydes, ketones, alcohols, and among other compounds that can generate “off-note”.

[0044] Examples of compounds generating “off-note” may include, but may not be limited to, volatile compounds (e.g., aldehydes, ketones, alcohols, alkaloids, phospholipids, phenolics), nonvolatile compounds (e.g., saponins), small peptides, free amino acids, or any combinations thereof. Such compounds may be responsible for contributing negative perception (e.g., bitterness, astringency) to the legume protein products.

[0045] The process comprises the steps of (a), purifying a legume protein containing material, optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; (b). treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and (c). removing the encapsulated flavor compounds to obtain the legume protein product. If unused encapsulating agents are present in the legume protein product, the process may then include an additional step of removing the unused encapsulating agents from the legume protein product.

[0046] The process may reduce content of one or more flavor compounds by at least 50%, preferably at least 90%, more preferably at least 98%, or even more preferably 100% in the resulting legume protein product as compared to an equivalent untreated legume protein product.

[0047] In one aspect, the legume protein product treated by the process has content of one or more flavor compounds reduced by at least 50%, preferably at least 90%, more preferably at least 98%, or even more preferably 100%, as compared to an equivalent untreated legume protein product.

[0048] In one aspect, at least 50%, preferably at least 90%, more preferably at least 98%, of one or more flavor compounds are removed in the legume protein product prepared by the process of the present disclosure; even more preferably, the legume protein product is completely free of any flavor compounds.

[0049] The starting material - legume protein containing material - can include legume protein with different concentrations (e.g., pulse, flour, concentrate, isolate, or intermediate products of the process described in the present disclosure) obtained from different botanical sources that may include, but may not be limited to, soy, fava, pea, chickpea, lupin, beans, mung bean, lentil, or any combinations thereof. Preferably, soy protein concentrate can be used as the starting material.

[0050] Preferably, the enzyme is lipase that may include, but may not be limited to, phospholipase A2 (PLA2), phospholipase Al (PLA1), phospholipase C (PLC), phospholipase D (PLD), or any combinations thereof.

[0051] Preferably, the encapsulating agent is cyclodextrin, that may include, but may not be limited to, alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, or any combinations thereof; more preferably, the cyclodextrin used can be beta-cyclodextrin.Process for increasing protein content of a legume protein product

[0052] The present disclosure provides a process for increasing protein content of a legume protein product.

[0053] The process comprises the steps of (a), purifying a legume protein containing material, optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; (b). treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and (c). removing the encapsulated flavor compounds to obtain the legume protein product. If unused encapsulating agents are present in the legume protein product, the process may then include an additional step of removing the unused encapsulating agents from the legume protein product.

[0054] The process may increase protein content of the resulting legume protein product by at least 10%, at least 20%, at least 25%, at least 50%, at least 75%, at least 90%, or at least 100% as compared to an equivalent untreated legume protein product.

[0055] The legume protein product treated by the process may have a protein content increased by at least 10%, at least 20%, at least 25%, at least 50%, at least 75%, at least 90%, or at least 100% as compared to an equivalent untreated legume protein product.

[0056] The starting material - legume protein containing material - can include legume protein with different concentrations (e.g., pulse, flour, concentrate, isolate, slurry, cake, low-protein stream, high-protein stream, or intermediate products of the process described in the present disclosure) obtained from different botanical sources that may include, but may not be limited to, soy, fava, pea, chickpea, lupin, beans, mung bean, lentil, or any combinations thereof. Preferably, soy protein concentrate can be used as the starting material.

[0057] Preferably, the enzyme is lipase that may include, but may not be limited to, phospholipase A2 (PLA2), phospholipase Al (PLA1), phospholipase C (PLC), phospholipase D (PLD), or any combinations thereof.

[0058] Preferably, the encapsulating agent is cyclodextrin, that may include, but may not be limited to, alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, or any combinations thereof; more preferably, the cyclodextrin used can be beta-cyclodextrin.Legume protein product

[0059] The legume protein product of the instant invention, prepared by any process described in the present disclosure, has one or more improved attributes as compared to an equivalent untreated legume protein product. Examples of an equivalent untreated legume protein product may include, but may not be limited to, a soy protein concentrate that has not been prepared or treated by any process described in the present disclosure.

[0060] In one aspect, one or more encapsulating agents are used in a process for preparing the legume protein product as described in the present disclosure. Upon completion of the process, most of the encapsulating agents may have been used to encapsulate flavor compounds present in the starting material, where some encapsulating agents may remain unused and be retrieved with the legume protein product; thus, the legume protein product may comprise one or more encapsulating agents. Preferably, the legume protein product may be completely free of any encapsulating agents.

[0061] In one aspect, the legume protein product described in the present disclosure may comprise one or more encapsulating agents, in which such encapsulating agents may be added in a process for preparing the legume protein product described in the present disclosure and may remainunused upon completion of such process (i.e., unused encapsulating agents). In other words, the legume protein product may comprise one or more unused encapsulating agents. Preferably, the legume protein product may not comprise any unused encapsulating agents.

[0062] Cyclodextrin is an example of the encapsulating agent, which may include, but may not be limited to, alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, or any combinations thereof; preferably, the cyclodextrin used can be beta-cyclodextrin.

[0063] The one or more attributes that are improved compared to an equivalent untreated legume protein product may include, but may not be limited to, a reduced flavor compound content, an increased protein content, or any combinations thereof.

[0064] Preferably, the legume protein product described in the present disclosure may have a reduced flavor compound content and an increased protein content compared to an equivalent untreated legume protein product. More preferably, the legume protein product described in the present disclosure may have a flavor compound content reduced by at least 50%, preferably at least 90%, more preferably at least 98%, even more preferably 100%, and a protein content increased by at least 10% compared to an equivalent untreated legume protein product.

[0065] In one aspect, the legume protein product described in the present disclosure may have a reduced flavor compound content compared to an equivalent untreated legume protein product; more preferably, the legume protein product may have off-notes partially or completely removed. In other words, the content of one or more flavor compounds in the legume protein product is reduced, preferably completely eliminated, as compared to an equivalent untreated legume protein product. Flavor compounds are substances present in legume protein products that impart one or more flavors (e.g., an earthy flavor, a savory flavor, a meaty flavor, a brothy flavor, a grainy flavor, a cereal flavor, a malty flavor, a toasted flavor, a beany flavor, a green flavor) to the legume protein product. Examples of the flavor compounds may include, but may not be limited to, phospholipid, phenolic, furan, pyran, organic acid, aldehyde, alcohol, ketone, pyrazine, lactone, thiol, sulfide, carboxylic acid, or any combinations thereof. Preferably, the flavor compounds may include, but may not be limited to, phospholipid, phenolic, aldehyde, ketone, alcohol, furan, or any combinations thereof.

[0066] Preferably, the legume protein product described in the present disclosure may have content of one or more flavor compounds reduced by at least 50%, preferably at least 90%, more preferably at least 98%, even more preferably 100%, as compared to an equivalent untreated legume protein product.

[0067] In one aspect, some flavor compounds in the legume protein product described in the present disclosure may be completely eliminated and non-detectable. Examples of such flavor compounds may include, but may not be limited to, l-penten-3-ol, pentanal, 1 -pentanol, 2-hexenal, 1 -hexanol, 2-heptanone, heptanal, 2-heptenal, l-octen-3-ol, 2-pentylfuran, l-octen-3- one, 2-nonenal, or any combinations thereof.

[0068] As used herein, the term “non-detectable” refers to an amount of a compound that cannot be quantified or detected by conventional analytical chemistry (e.g., use of gas chromatography (GC) / mass spectrometry (MS)).

[0069] In one aspect, content of some flavor compounds in the legume protein product described in the present disclosure may be reduced by at least 98% as compared to an equivalent untreated legume protein product. Examples of such flavor compounds may include, but may not be limited to, l-penten-3-ol, pentanal, 1 -pentanol, hexanal, 2-hexenal, 1 -hexanol, 2-heptanone, heptanal, 2- heptenal, l-octen-3-ol, 2-pentylfuran, l-octen-3-one, 2-nonenal, or any combinations thereof.

[0070] In one aspect, content of some flavor compounds in the legume protein product described in the present disclosure may be reduced by 90% as compared to an equivalent untreated legume protein product. Examples of such flavor compounds may include, but may not be limited to, 1- penten-3-ol, pentanal, 1 -pentanol, hexanal, 2-hexenal, 1 -hexanol, 2-heptanone, heptanal, 2- heptenal, l-octen-3-ol, 2-pentylfuran, l-octen-3-one, 2-nonenal, or any combinations thereof.

[0071] In one aspect, content of some flavor compounds in the legume protein product described in the present disclosure may be reduced by 50% as compared to an equivalent untreated legume protein product. Examples of such flavor may include, but may not be limited to, l-penten-3-ol, pentanal, 1 -pentanol, hexanal, 2-hexenal, 1 -hexanol, 2-heptanone, heptanal, 2-heptenal, benzaldehyde, l-octen-3-ol, 2-pentylfuran, l-octen-3-one, nonanal, 2-nonenal, or any combinations thereof.

[0072] The legume protein product described in the present disclosure may have an aldehyde content reduced, by at least 50%, as compared to an equivalent untreated legume protein product. The aldehyde may include, but may not be limited to, pentanal, hexanal, 2-hexenal, heptanal, 2- heptanal, benzaldehyde, nonanal, 2-nonenal, or any combinations thereof. Aldehydes detectable by humans upon tasting may be so potent that they cannot be identified or quantified by conventional analytical chemistry and yet be decreased by the process of this invention.

[0073] The legume protein product described in the present disclosure may have a ketone content reduced, by at least 50%, as compared to an equivalent untreated legume protein product. The ketone may include, but may not be limited to, 2-heptanone, l-octen-3-one, or any combinations thereof. Ketones detectable by humans upon tasting may be so potent that they cannot be identified or quantified by conventional analytical chemistry and yet be decreased by the process of this invention.

[0074] The legume protein product described in this disclosure may have an alcohol content reduced, by at least 50%, as compared to an equivalent untreated legume protein product. Thealcohol may include, but may not be limited to, l-penten-3-ol, 1 -pentanol, 1 -hexanol, l-octen-3- ol, or any combinations thereof. Alcohols detectable by humans upon tasting may be so potent that they cannot be identified or quantified by conventional analytical chemistry and yet be decreased by the process of this invention.

[0075] The legume protein product described in the present disclosure may have a furan content reduced, by at least 50%, as compared to an equivalent untreated legume protein product. The furan may include, but may not be limited to, 2-pentylfuran. Furans detectable by humans upon tasting may be so potent that they cannot be identified or quantified by conventional analytical chemistry and yet be decreased by the process of this invention.

[0076] Preferably, the legume protein product described in the present disclosure may be free of and / or have a non-detectable amount of one or more flavor compounds, where the flavor compound may include, but may not be limited to, l-penten-3-ol, pentanal, 1 -pentanol, 2-hexenal, 1 -hexanol, 2-heptanone, heptanal, 2-heptenal, l-octen-3-ol, 2-pentylfuran, l-octen-3-one, 2- nonenal, or any combinations thereof.

[0077] In one aspect, the legume protein product described in the present disclosure may have an increased protein content as compared to an equivalent untreated legume protein product. Preferably, the legume protein product may have a protein content increased by at least 10% as compared to an equivalent untreated legume protein product.

[0078] In one aspect, attributes other than flavor compound content and protein content of the legume protein product described in the present disclosure can be improved compared to an equivalent untreated legume protein product. Such attributes may include, but may not be limited to, taste, aroma, color, free amino acid concentration, metal ion concentration, carbohydrate concentration, or any combinations thereof.

[0079] In one aspect, the legume protein product described in the present disclosure can be present with different concentrations (e.g., pulse, flour, concentrate, or isolate). The legume protein product can undergo one or more post-treatment processes to be transformed into a desirable form (e.g., an extrudate), in which the post-treatment process may include, but may not be limited to, extrusion.Food product

[0080] The legume protein product described in the present disclosure can be used to prepare a food product, which may include, but may not be limited to, a meat substitute product, a dairy substitute product, a meat and / or dairy containing product, a bakery product, a beverage product, products designed for sports and / or for health and nutrition (such as shakes, high-protein dairymeals, meal replacements) that may or may not be vegan / vegetarian, or a convenience food product.

[0081] Meat substitute products are also referred to meat alternative products, meat analogue products, meat mimicking products, meat replacement products, and the like. By “meat” it is meant herein not only substitutes for red meats (from Bovidae), such as beef, lamb, bison, goat, and mutton, but any animal meat, including poultry (e.g., chicken, turkey, duck, ostrich, pigeon), fish (e.g., whether farmed or wild-caught), and shellfish (e.g., shrimp, prawns, crab, crayfish, lobster, scallops).

[0082] Alternatively, the meat substitute product of the present invention is a pet food. The term “pet food” means any food composition intended to be consumed by a pet. A pet may be any domestic or tamed animal kept for companionship or pleasure. The pet can, preferably, be an avian, bovine, canine, equine, feline, hircine, lupine, murine, ovine, or porcine animal. Preferably, the pet is a dog or a cat.

[0083] Preferred meat substitute products are vegetarian or vegan seafood alternatives, poultry alternatives, but also burger patties, sausages, meatballs, minced meat, cold-cuts, nuggets, crumbles, breakfast meat, or meat-like toppings (e.g., for dishes such as pizzas, pies, flans, quiche, and the like). By mincemeat it is included herein also toppings for dishes such as pizzas, pies, flans, quiche, and the like.

[0084] Also disclosed herein is a meat extender food product that comprises the meat substitute product described in the present disclosure and one or more meat derived ingredients. Meat derived ingredients may include any animal protein such as minced meat and meat cut-offs.

[0085] As used herein, by “vegetarian” it is meant not comprising any animal meat products, including bovid meats, poultry, fish, crustaceans, mollusks, game, and the like.

[0086] As used herein, by “vegan” it is meant not comprising any animal meat products, nor any animal by-products, the latter referring to products such as eggs, honey, and dairy products.

[0087] As used herein, the term “meat extender food product” refers to a food product containing meat derived ingredients, in which a part of the meat derived ingredients has been replaced by a non-meat substance having a substantial protein content.

[0088] The meat substitute products can be frozen, fresh, or canned. The meat substitute products can be sold cooked, pre-cooked, or raw. The meat substitute products can undergo lethality cook step(s) to make the product fully cooked and ready to eat.

[0089] Dairy substitute products also refer to dairy alternative products, dairy analogue products, dairy mimicking products, dairy replacement products, and the like.

[0090] In one aspect, the dairy substitute products that can be made from the legume protein product described in the present disclosure may include, but are not limited to, dairy alternativesof cream cheese products, table cream products, sour cream products, heavy cream products, whipping cream products, light cream products, coffee cream products, spreadable cheese products, or spreadable processed cheese products. The dairy substitute products can be directly consumed (e.g., ready to use or ready to consume application) and / or used in savory or sweet cooking preparation. For example, the dairy substitute product can be applied with fruits and / or salads.Examples

[0091] The invention is further described in detail by reference to the following experimental examples. These examples are provided for purposes of illustration only and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.Example 11.1. Materials and Method

[0092] This method uses a combination of protein precipitation, enzyme-treatment, molecular- inclusion, and ultrasound technology, to remove off-flavors and their precursors from soy protein material such as soy protein concentrate (SPC).1.1.1 Pre-treatment without Ultrasonic

[0093] SPC samples were diluted at 50g / L in pure water and pH of the solution was adjusted to 8.0 with 2M NaOH aqueous solution. The resulting samples were incubated at 43°C with stirring at 200 RPM / min for 1 hour. Afterwards, the samples were acidified to pH 4.5 with a 4M HC1 aqueous solution. After protein precipitation, the recovered samples were centrifuged at 5000 RPM / min for 5 minutes and the supernatant was removed. An aliquot of the supernatant was stored for later analysis. The pellets were rinsed with 200mL of acidified water (pH 4.5 - adjusted with 4M HC1) and centrifuged at 5000 RPM / min for 5 minutes to remove the supernatant. The recovered pellets were diluted at 50g / L in pure water and pH of the resulting samples was adjusted to 8.0 with 2M NaOH aqueous solution.

[0094] Next, 90 ppm of phospholipase was added, and the mixture was incubated at 43 °C under stirring at 200 RPM / min for 90 minutes. Phospholipids in the samples were then hydrolyzed by phospholipase into lysophosphatides, lipophilic substances (LyP), and free fatty acids (FFA) in the mixture. Then, 12mM of beta-cyclodextrin (PCD) was added and the mixture was incubatedfor 60 minutes. PCD can form molecular-inclusion complexes with LyP and FFA such that LyP and FFA were encapsulated by PCD.

[0095] After treatment with PCD, the sample was acidified to pH 4.5 with a 4M HC1 solution and centrifuged at 5000 RPM / min for 5 minutes. Pellets were washed twice with 200mL of acidified water (pH 4.5 - adjusted with 4M HC1) and centrifuged at 5000 RPM / min for 5 minutes. The recovered protein pellets were collected and dried in an oven at 40°C to obtain the soy protein concentrate with reduced off-notes (treated samples).1.1.2 Pre-treatment with Ultrasonic

[0096] Alternatively, ultrasound treatment can be applied to further remove phospholipids and other off-note precursor molecules (e.g., phenolics and flavonoids) from the soy protein material. SPC samples were diluted at 50 g / L in pure water and pH of the solution was adjusted to 8.0 with 2M NaOH aqueous solution. The resulting samples were subjected to ultrasonic treatment for 0, 2.5, 10, 20, and 40 minutes at 2,500 W and with the sound frequency from 20 to 60 kHz The ultrasonic-treated samples were incubated at 43 °C with stirring at 200 RPM / min for 1 hour. Afterwards, the samples were acidified to pH 4.5 with a 4M HC1 aqueous solution. After protein precipitation, the recovered samples were centrifuged at 5000 RPM / min for 5 minutes and the supernatant was removed. An aliquot of the supernatant was stored for later analysis. The pellets were rinsed with 200 mL of acidified water (pH 4.5 - adjusted with 4M HC1) and centrifuged at 5000 RPM / min for 5 minutes to remove the supernatant. The recovered pellets were diluted at 50 g / L in pure water and pH of the resulting samples was adjusted to 8.0 with 2M NaOH aqueous solution.

[0097] Then, the samples were divided into two sets in which the first set of samples were treated with 1 mL of phospholipase enzyme and 12 mM of PCD, while the second set was not treated with phospholipase or PCD. Next, both sets of samples were incubated at 43°C at 150 RPM / min for 3 hours.

[0098] After treatment, the samples were acidified to pH 4.5 with a 4 M HC1 solution and centrifuged at 5000 RPM / min for 5 minutes. The pellets were washed twice with 200 mL of acidified water (pH 4.5 - adjusted with 4 M HC1) and centrifuged at 5000 RPM / min for 5 minutes. The recovered pellets were collected and dried in an oven at 40°C to obtain the soy protein concentrate with reduced off-notes (treated samples).1.2 Phospholipids and Phenolic Compounds Quantification

[0099] The phosphorus content of the sample was determined based on the standard curve of monobasic potassium phosphate, KH2PO4. The phospholipid content was determined bymultiplying the phosphorus content by a factor based on the average molecular weight of soybean phospholipids.

[0100] The phospholipid content was expressed in mg / 100 g of soy protein concentrate. The content of phospholipids removed from the sample was measured by the differences between the values obtained for starting SPC and the final treated product. The measurements were made in duplicate.

[0101] The results for phospholipid content and phenolic content were expressed as mean ± standard deviation. Significant differences between the treatments in the comparison experiment were analyzed by ANOVA analysis of variance with p < 0.05 (phospholipids and phenolics removal from SPC). The means of the treatments were compared by Duncan’s test (p < 0.05). All analyses were carried out in duplicates.

[0102] Total compounds were quantified using the Folin-Ciocalteau spectrophotometric method, using gallic acid as a reference standard (R2= 0.9944). The absorbance was measured at 760 nm using a spectrophotometer. The results were expressed as mg of gallic acid equivalent per 100 g of sample (mg EAG / 100 g).1.3 Results and Discussion1.3.1 Pre-treatment without Ultrasonic

[0103] A validation experiment was carried out based on the combination of the factors including enzyme concentration (10 ppm) and PCD concentration (8 mM) for removing off-notes (e.g., phospholipids and phenolics). Control trials (2 to 4) were also carried out without the addition of phospholipase and / or PCD. The results for the removal of the phospholipid and phenolic content are presented in Table 1.Table 1* = glycolipase; ** = phospholipase

[0104] The results show that removals of phospholipids and phenolics is a function of concentration of the enzyme, concentration of cyclodextrin, and any combinations thereof.

[0105] Further, it can be observed from Table 1 that treatment with both phospholipase and PCD (i.e., Trial 1 or 7) has the most significant effects on the removals of phospholipids and phenolics as compared to the treatment without the use of phospholipase and PCD (i.e., non-treatment in Trial 2). Also, treatment with phospholipase alone (i.e., Trial 5 or 6) or with PCD alone (i.e., Trial 3) seems to have different effects on the removals. Thus, treatment with both phospholipase and PCD would maximize the removals of both phospholipids and phenolics.1.3,2 Pre-treatment with Ultrasonic

[0106] The kinetic studies showed a positive influence on the use of ultrasonic treatment to remove flavor compounds from SPC. For example, the yield of phospholipid removal in the samples with ultrasonic pre-treatment (125% yield) was about 2.5 times higher than the yield of phospholipid removal in the samples without ultrasonic pre-treatment (52% yield).Example 22.1 Materials and Method

[0107] 0.5 g of the treated samples was weighed directly in 20 mL vials for headspace solid-phase microextraction (Headspace- SPME) and subjected to heating for 60 minutes. The heated samples were then inserted to a gas-chromatography coupled to mass spectrometer (GCMS, Shimadzu) using SPME fiber assembly Carboxen / Polydimethylsiloxane (CAR / PDMS). Areas of the detected volatile compounds were calculated and compared among different treatment conditions. Identification of the detected volatile compounds present in the samples was performed using the mass spectra library (NIST database).2.2 Results and Discussion

[0108] As observed from Table 2, fifteen of the detected volatile compounds were identified and the treatment with phospholipase and PCD of the instant invention significantly reduced concentration of these identified volatile compounds. Some of these identified volatile compounds were completely eliminated by the treatment as they were non-detected in the measurements. Ultrasonic pre-treatment was also shown to increase the efficacy of reduction / removal of the volatile compounds.

[0109] A further observation from Table 2 revealed that the off-notes associated with these fifteen identified volatile compounds were significantly removed or completely eliminated in the treated soy protein product.Table 2Example 33.1 Materials and Method

[0110] A study was carried out to compare the sensory attributes of sausage substitutes made from soy protein products prepared by processes described in the present disclosure (i.e., preparation process without ultrasonic pre-treatment (treated without ultrasound) and preparation process with ultrasonic pre-treatment (treated with ultrasound)) with a sausage substitute made from an equivalent untreated soy protein product. Recipes of such sausage substitutes are shown in Table 3. Amounts of the ingredients are measured in wt% of the total weight of the sausage substitutes.Table 3

[0111] Following steps were taken to prepare the sausage substitute.

[0112] (1). Ingredients in Table 3 were weighed and mixed to form an emulsion.

[0113] (2). The emulsion was transferred to a grinder to form a sausage substitute.

[0114] (3). The sausage substitute was then dried at 55°C for 20 minutes.

[0115] (4). The heated sausage substitute was then steam-cooked at 80°C for 15 minutes.

[0116] (5). The cooked sausage substitute was cooled for further use.3.2 Sensory evaluation

[0117] An informal sensory evaluation was conducted to compare the sensory attributes of the different sausage substitutes of Table 3. Samples of the sausage substitutes were tasted independently by an informal panel of about 120 panelists for different attributes (e.g.,appearance, flavor, taste). Sensory evaluation was performed on boiled sausage substitutes according to Kelly’s repertory grid method in which rate-all-that-apply (RATA) and variation of check-all-that-apply (CATA) were applied for consumer characterization. Samples of the sausage substitutes were tasted independently by 120 panelists for different attributes (e.g., appearance, flavor, taste, and texture) and global evaluation. Scores, representing a stronger intensity / liking for their opinions on color, appearance, odor, taste, texture, and overall acceptability, were assigned using a hedonic scale as follows: Like extremely-9; Like very much-8; Like moderately- 7; Like slightly-6; Neither like nor dislike-5; Dislike slightly-4; Dislike moderately-3; Dislike very much-2; and Dislike extremely-1.

[0118] First, the overall acceptability test results showed that there were no differences between the treated samples and the in terms of consumer liking (Table 4) which may be an indication of a good fit for the sausage formulation since there were no differences in terms of appearance and aroma.Table 4

[0119] Scores for each sensory attribute were obtained and analyzed via analysis of variance (95% confidence interval). Additionally, Partial Least Squares Regression (PLSR) was utilized, employing the mean acceptance ratings of the sausage flavor as the dependent variable (Y) and the mean intensities of the attributes as independent variables (X). PLSR was employed to reduce data dimensionality and select the best variables based on covariance capable of explaining the acceptance ratings. In essence, PLSR reduced the independent explanatory variables (sensory attributes) to a smaller set of predictors that were then used to perform regression. Furthermore, Variable Importance in Projections (VIPs) was determined for each of the explanatory variables; unlike loadings and scores, VIPs were evaluated considering the entire model such that the VIP was the sum of all contributions of that variable across components, based on the variable's weight in each component combined with the variation captured in that component relative to other components. Two attributes were deemed significant in the VIP analysis: "like-sausage taste" and "like-crude grain taste," with the former having a positive association / effect on the acceptance of sausage samples and the latter a negative association / effect. Higher intensities of sausage flavor were associated with positive impacts on sausage acceptance. As both treated samples (treatedwith and without ultrasound) scored higher than the untreated samples in the "like-sausage taste" attribute, they exhibited stronger intensities of sausage flavor and acceptance scores. On the other hand, increased intensity of the "like-crude grain taste" imparted a more drastic and negative impact on the overall acceptance of the sausage substitutes (Tables 5 and 6). As both treated samples scored lower than the untreated samples in the "like-crude grain taste" attribute, they displayed a decreased intensity of like-crude grain taste, and thus a less drastic and negative impact on the flavor of sausage substitutes, compared to the untreated samples.

[0120] In summary, the treatment of the samples, including both with and without ultrasound, demonstrated a contribution to a positive effect on the flavor of the sausage substitutes.Table 5Table 6

Claims

CLAIMS1. A process for preparing a legume protein product, comprising the steps of: a. providing a legume protein containing material; b. purifying the legume protein containing material to obtain a purified material; c. treating the purified material with one or more enzymes, followed with one or more encapsulating agents, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and d. removing the encapsulated flavor compounds from the mixture of treated components to obtain the legume protein product; wherein the legume protein containing material is optionally treated with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes; and wherein the legume protein product has a reduced flavor compound content and / or an increased protein content compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (d).

2. The process of claim 1, wherein the enzyme and encapsulating agent are not simultaneously added to the purified material.

3. The process of any of the preceding claims, wherein the legume protein containing material comprises legume protein selected from the group consisting of soy protein, fava protein, pea protein, chickpea protein, lupin protein, beans protein, mung bean protein, lentil protein, and any combinations thereof.

4. The process of any of the preceding claims, wherein the enzyme is a phospholipase selected from the group consisting of phospholipase A2 (PLA2), phospholipase Al (PLA1), phospholipase C (PLC), phospholipase D (PLD), and any combinations thereof.

5. The process of any of the preceding claims, wherein the encapsulating agent is selected from the group consisting of cyclodextrin, starch, starch derivatives, cellulose, cellulose derivatives, plant exudates, plant extracts, marine extracts, microbial polysaccharides, animal polysaccharides, and any combinations thereof.

6. The process of any of the preceding claims, wherein the cyclodextrin is selected from the group consisting of alpha-cyclodextrin, beta-cyclodextrin, gamma-cyclodextrin, and any combinations thereof.

7. The process of any of the preceding claims, wherein the flavor compounds are selected from the group consisting of phospholipid, phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.

8. A process for reducing flavor compound content of a legume protein product, comprising the steps of:a. purifying a legume protein containing material, optionally treating with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes, to obtain a purified material; b. treating the purified material with phospholipase, followed with cyclodextrin, to obtain a mixture of treated components of encapsulated flavor compounds and the legume protein product; and c. removing the encapsulated flavor compounds to obtain the legume protein product; wherein the legume protein product has a flavor compound content reduced by at least 50%, preferably at least 90%, more preferably at least 98%, even more preferably 100%, compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).

9. The process of claim 8, wherein the flavor compounds are selected from the group consisting of phospholipid, phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.

10. The process of any of claims 8 to 9, wherein the legume protein product has an aldehyde content, a ketone content, an alcohol content, and / or a furan content reduced by at least 50% compared to an equivalent untreated legume protein product that has not been subject to the process having steps (a) to (c).

11. A legume protein product prepared by the process of any of the preceding claims or by a process comprising the steps of: a. providing a legume protein containing material; b. purifying the legume protein containing material to obtain a purified material, optionally treating the legume protein containing material with ultrasound for an ultrasonic-treatment period from 20 to 40 minutes; c. adding one or more enzymes, followed by one or more encapsulating agents, to the purified material to obtain a mixture of treated components; and d. removing encapsulated flavor compounds from the mixture of treated components to obtain the legume protein.

12. The legume protein product of claim 11 comprising one or more encapsulating agents.

13. The legume protein product of any of claims 11 to 12 being free of any encapsulating agents.

14. The legume protein product any of claims 11 to 13, wherein the encapsulating agent is selected from the group consisting of cyclodextrin, starch, starch derivatives, cellulose, cellulose derivatives, plant exudates, plant extracts, marine extracts, microbial polysaccharides, animal polysaccharides, and any combinations thereof.

15. The legume protein product of any of claims 11 to 14, having content of one or more flavor compounds reduced by at least 50%, preferably at least 90%, more preferably at least 98%,even more preferably 100%, as compared to an equivalent untreated legume protein product which has not subject to the process of any of claims 1 to 10.

16. The legume protein product of any of claims 11 to 15, wherein the flavor compounds are selected from the group consisting of phospholipid, phenolic, aldehyde, ketone, alcohol, furan, and any combinations thereof.

17. The legume protein product of any of claims 11 to 16, being free of and / or having a non- detectable amount of one or more flavor compounds selected from a group consisting of 1- penten-3-ol, pentanal, 1 -pentanol, 2-hexenal, 1 -hexanol, 2-heptanone, heptanal, 2-heptenal, l-octen-3-ol, 2-pentylfuran, l-octen-3-one, 2-nonenal, and any combinations thereof.

18. The legume protein product of any of claims 11 to 17, having an aldehyde content, a ketone content, an alcohol content, and / or a furan content reduced by at least 50% compared to an equivalent untreated legume protein product that has not been subject to the process of any of claims 1 to 10.

19. The legume protein product of any of claims 11 to 18, having a protein content increased by at least 10% as compared to an equivalent untreated legume protein product.

20. A food product comprising the legume protein product of any of claims 11 to 19 and other food ingredients, preferably wherein the food product is selected from a group consisting of meat substitute products, dairy substitute products, bakery products, beverage products, and convenience food products.

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