Plant protein isolates and use thereof

Abstract

The present invention is directed to plant protein isolates comprising a first protein fraction characterized by a molecular weight between 5kDa and 17kDa, and a second protein fraction characterized by a molecular weight between 85kDa and 105kDa, and methods of use thereof as whitening agents of food products.

Description

PLANT PROTEIN ISOLATES AND USE THEREOFFIELD OF THE INVENTION

[0001] The present invention relates to compositions of plant protein isolates and methods of use thereof as whitening agents of food products.BACKGROUND OF THE INVENTION

[0002] Plant-based proteins are widely exploited for a total or partial replacement of chemicals and animal proteins in human nutrition. As consumer awareness of the negative impacts of chemicals grows, the food industry is turning to ‘clean-label’ food products, which have minimal to no synthetic chemicals. Titanium dioxide (TiO2; E171) is widely used as a whitening food additive in a variety of food products, including bakery products, sauces, cheeses, edible ices, and sweets. Recently, its use as a food additive has been reevaluated by the regulatory authorities due to raised concerns for genotoxicity. Proteins derived from plant extracts can offer a natural alternative for chemical food additives as they are naturally digestible, breaking down into amino acids within the human gastric system, thereby posing no health risks due to toxicity. The properties of plant protein extracts produced by different methods are being evaluated for determining their potential applicability in industrial processes, including in the food industry.

[0003] Kornet et al., 2021 (Food Hydrocolloids 120 (2021) 106891) explored the gelling behavior of a pea protein concentrate, an albumin-fraction and a globulin-rich fraction, in comparison with and as substitute for whey protein isolate. Kornet et al., 2022 (Food Hydrocolloids 125 (2022) 107427) disclose that fractionation methods affect the gelling properties of pea proteins in emulsion-filled gels. Plant enzymes are typically used as commercially available legume flours (e.g., enzyme-active soybean flour). Another means for introducing enzymes such as lipoxygenases to bakery products is through the inclusion of an engineered yeast able to produce and secrete the active enzyme during the production process. EP 1638405 relates to a method for bleaching or whitening a dairy product, comprising adding a lipoxygenase to the dairy product, preferably a soy lipoxygenase. WO 2023 / 139590 relates to colorants of a natural origin having improved physical-chemicalproperties, such as light and thermal stability, compositions and articles comprising and / or made of such colorants.

[0004] Pea-based proteins are widely used in food as a plant alternative to animal proteins. These mainly include pea globulin fractions which constitute 65-80% of pea-protein isolates. Pea albumin fractions which constitute 10-25% of pea-protein isolates received less attention and are often considered by-products of a globulin fractionation process, that are not suitable for use in food products.

[0005] Pea globulin fractions can be classified into legumin, vicilin and convicilin, although the latter is sometimes considered part of the vicilin subgroup. At neutral pH, legumin is mostly present as a hexamer with a molecular weight of 320-380 kDa, while vicilin and convicilin are mostly present as trimers with molecular weights of 170 kDa and 290 kDa, respectively. Pea albumins refer to a class of proteins, including PAI (~6 kDa), PA2 (~26 kDa), lectin (~17 kDa), protease inhibitors, and lipoxygenase (97-100 kDa).

[0006] A commonly reported procedure for fractionating plant proteins involves aqueous fractionation, which includes a two-stage process: a solubilization step at alkaline pH followed by a precipitation step of globulin proteins at acidic pH. The precipitated globulins are then separated from the albumin-containing aqueous fraction by centrifugation. Direct albumin isolation from pea flour can be prepared by selective extraction procedures, based on Osborne protein classification.

[0007] However, this method is highly deficient, since it inevitably results in crosscontamination between albumin and globulin fractions, rendering it practically impossible to isolate an almost pure albumin fraction. Furthermore, separation of the precipitated globulins from the albumin-containing slurry by centrifugation or filtration is complicated thereby impeding the scale-up of the selective extraction methods to industrial level.

[0008] To this end, there is a great need for a new method for highly selective plant albumin extraction. Moreover, conventional protein isolates have an off-white or brown appearance (due to various plant by-products) and are further characterized by an unpleasant off-taste.

[0009] Accordingly, there is a great unmet need for white-colored plant extracts with improved taste, concomitantly with desirable nutritional features, such as high levels of essential amino acids, and low lipid levels, for application in the food industry. There is also an unmet need for whitening additives in the food industry that are eco-friendly, sustainable, and safe as alternatives to chemical food colorants.SUMMARY OF THE INVENTION

[0010] The present invention provides compositions comprising protein isolates derived from legume extracts which are characterized by a total color difference (AE) in CIELAB coordinates of less than 4, relative to titanium dioxide powder. The compositions are useful as food whitening additives.

[0011] The present invention is based, in part, on the unexpected finding of compositions comprising protein isolates derived from legume extracts which are characterized by a white color, similar to the TiCh whitening food additive. The compositions can be safely consumed as natural alternatives to TiCh whitening agent without inflicting any health risks to the consumers.

[0012] According to a first aspect, the present invention provides a food whitening additive comprising a plant protein isolate derived from a legume extract, wherein the plant protein isolate has a protein content of at least 80% and is characterized by: (i) a first protein fraction having a molecular weight between 5 kDa and 17 kDa, (ii) a second protein fraction having a molecular weight between 85 kDa and 105 kDa in a weight portion relative to the total plant proteins of the plant protein isolate of at least 1%, and (iii) a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction, of 2.25 or less, and wherein the plant protein isolate is further characterized by a total color difference (AE) in CIELAB coordinates of less than 4, relative to titanium dioxide powder.

[0013] In one embodiment, the plant protein isolate is characterized by L* CIELAB value in a range between about 90 and about 95; a* CIELAB value in a range between about (- 0.1) and about 0.4; and b* CIELAB value in a range between about 4.0 and about 10.0, including each value within the specified ranges. In another embodiment, the plant proteinisolate is characterized by L* CIELAB value in a range between about 80 and about 95; a* CIELAB value in a range between about (-2) and about 1.5; and b* CIELAB value in a range between about 3 and about 10.0, including each value within the specified ranges.

[0014] In some embodiments, the plant protein isolate is in the form of a powder (also referred to herein as a powderous composition). In other embodiments, the plant protein isolate (e.g., in the form of a powder) is characterized by Dv(50) between about 10 pm and about 20 pm, including each value within the specified range. In yet other embodiments, the plant protein isolate (e.g., in the form of a powder) is characterized by Dv(50) between about 35 pm and about 55 pm, including each value within the specified range.

[0015] In certain embodiments, the plant protein isolate (e.g., in the form of a powder) is characterized by Dv(90) between about 30 pm and about 50 pm; Dv(10) between about 1 pm and about 5 pm; and Dv(50) between about 10 pm and about 20 pm, including each value within the specified ranges. In other embodiments, the plant protein isolate (e.g., in the form of a powder) is characterized by Dv(90) between about 100 pm and about 150 pm; Dv(10) between about 4 pm and about 10 pm; and Dv(50) between about 35 pm and about 55 pm, including each value within the specified ranges. In particular embodiments, the plant protein isolate (e.g., in the form of a powder) is characterized by at least one of: uniformity of between about 0.6 and about 1.2; span of between about 2 and about 3.7; D[3,2] of between about 4 pm and about 9 pm; and D[4,3] of between about 15 pm and about 25 pm, including each value within the specified ranges. In other particular embodiments, the plant protein isolate (e.g., in the form of a powder) is characterized by at least one of: D[3,2] of between about 15 pm and about 20 pm; and D[4,3] of between about 50 pm and about 65 pm, including each value within the specified ranges.

[0016] In further embodiments, the plant protein isolate in the form of a powder is characterized by a specific surface area of at least 300 m2 / kg. In various embodiments, the plant protein isolate in the form of a powder is characterized by a specific surface area of at least 700 m2 / kg.

[0017] In specific embodiments, the plant protein isolate is derived from seeds of a legume plant.

[0018] In other embodiments, the legume is selected from the group consisting of pea, soy, chickpea, lentil, bean, peanut, and any combination thereof. Each possibility represents a separate embodiment. In specific embodiments, the legume is selected from the group consisting of pea, soy, chickpea, and lentil. Each possibility represents a separate embodiment. In one embodiment, the legume is green pea. In another embodiment, the legume is yellow pea. In yet another embodiment, the legume is red lentil. In a further embodiment, the legume is soy.

[0019] In some embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 2%. In other embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 3%. In yet other embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 5%. In additional embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 10%. In certain embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 12.5%. In particular embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 15%. In specific embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 20%.

[0020] In further embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa comprises a lipoxygenase. In one embodiment, the lipoxygenase is enzymatically inactive. In additional embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa comprises a combination of 9s-lipoxygenase-3 and linoleate 9s-lipoxygenase-2. In specific embodiments, the second protein fraction having a molecular weight between 85 kDa and 105 kDa consists essentially of a combination of 9s-lipoxygenase-3 and linoleate 9s-lipoxygenase-2. In one embodiment,the weight ratio between the 9s-lipoxygenase-3 and the linoleate 9s-lipoxygenase-2 is in the range between about 3: 1 and about 2: 1, including each value within the specified range.

[0021] In some embodiments, the plant protein isolate has an albumin content of at least about 20% w / w. In additional embodiments, the plant protein isolate has an albumin content of at least about 30% w / w. In certain embodiments, the plant protein isolate has an albumin content of at least about 40% w / w. In other embodiments, the plant protein isolate is substantially devoid of globulins. In yet other embodiments, the plant protein isolate is substantially devoid of the reference protein fraction.

[0022] In various embodiments, the plant protein isolate is water-insoluble. In further embodiments, the plant protein isolate is capable of forming a suspension in an aqueous medium.

[0023] In additional embodiments, the plant protein isolate comprises trace amounts of a hydrocolloid.

[0024] In some embodiments, the present invention provides an edible composition comprising the food whitening additive disclosed herein. In one embodiment, the edible composition comprises at least 0.1 w / w of the food whitening additive disclosed herein. In further embodiments, the edible composition is in the form of a water-in-oil emulsion or an oil-in-water emulsion. Each possibility represents a separate embodiment. In additional embodiments, the edible composition is in the form of an oil-in-water emulsion comprising 0.03-3% w / w of the food whitening additive disclosed herein; 20%-98% w / w of an oily phase; and 2-80% w / w of an aqueous phase, including each value within the specified ranges.

[0025] In various embodiments, the present invention provides a food product comprising between 0.1 and 30% w / w of the food whitening additive disclosed herein. In certain embodiments, the food product comprises between 0.1 and 5% w / w of the food whitening additive disclosed herein.

[0026] In particular embodiments, the present invention provides a method of whitening a food product, comprising combining the product or a premix or intermediate thereof with the food whitening additive or the edible composition disclosed herein. In other particularembodiments, the food product is suitable for human consumption or for non-human animal consumption. Each possibility represents a separate embodiment.

[0027] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and / or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

[0028] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE FIGURES

[0029] Figures 1A-1B: (1A) An image of SDS-PAGE demonstrating the distribution of proteins according to their molecular weight in the Gavan green pea protein (GPP) extract, compared to a “full extract” extracted at pH 9.0, and the albumin fraction obtained by globulins precipitation. (IB) An image of SDS-PAGE demonstrating the distribution of proteins according to their molecular weight in the Gavan GPP isolate, compared to two commercial pea protein isolates (commercial 1 corresponds to a pea protein isolate (85%) of Dormeco Ltd.; and commercial 2 corresponds to a pea protein isolate (80%) of Pisumpro- 800).

[0030] Figures 2A-2B: Images demonstrating the whitening property of the Gavan GPP powder (right) compared to a commercial pea protein powder (left; 2A), and aqueous solutions consisting of 5% w / v Gavan GPP powder (right) compared to aqueous solutions consisting of 5% w / v commercial pea protein 85 (middle) and to 5% w / v commercial soy protein 90 (left; 2B).

[0031] Figure 3: A diagram of color coordinates measurements using HunterLab EasyMatchQC software showing L*, a*, and b* CIELAB values of Gavan GPP powder (“4”) compared to commercially available pea protein isolates of global brands and manufactures (“1”, “2”, and “3”).

[0032] Figure 4: A diagram of color coordinates measurements using HunterLab EasyMatchQC software showing L*, a*, and b* CIELAB values of Gavan GPP powder (“4”), and Gavan red lentils protein (“3”) powder compared to a commercial pea protein powder (“2”), a commercial soy protein powder (“1”), and a reference TiO2 powder (“5”). Delta E values are calculated as compared to the reference TiO2 powder.

[0033] Figures 5A-5B: Images of histograms demonstrating a particle size distribution analysis using Master Sizer 2000 by Malvern. (5A) Particle size before sonication of the Gavan GPP powder. (5B) Particle size after 5 seconds of sonication of the Gavan GPP powder. Results are presented as volume density (%) as a function of size (pm).

[0034] Figure 6: An image of SDS-PAGE demonstrating the distribution of proteins according to their molecular weight in the Gavan soy protein (SP) isolate, compared to a commercial soy protein isolate (“Solpro 922 isolate”).

[0035] Figures 7A-7B: Images of histograms demonstrating the particle size distribution analysis using Master Sizer 2000 by Malvern. (7A) Particle size before sonication of the Gavan SP powder. (7B) Particle size after 5 seconds of sonication of the Gavan SP powder.

[0036] Figure 8: An image of SDS-PAGE demonstrating the distribution of proteins according to their molecular weight in the Gavan red lentils isolate, compared to a full extract of red lentils proteins.

[0037] Figure 9: A diagram of color coordinates measurements using HunterLab EasyMatchQC software showing L*, a*, and b* CIELAB values of Gavan RLP powder samples.

[0038] Figures 10A-10B: Images of histograms demonstrating the particle size distribution analysis using Master Sizer 2000 by Malvern. (10A) Particle size before sonication of the Gavan RLP powder. (10B) Particle size after 5 seconds of sonication of the Gavan RLP powder.DETAILED DESCRIPTION OF THE INVENTION

[0039] The present invention in some embodiments thereof, refers to plant-derived protein compositions (or isolates) characterized by white color (almost identical to that of titanium dioxide), devoid of off-taste and characterized by improved organoleptic properties compared to conventional isolates. The plant-protein isolates of the invention have an increased albumin and total protein contents, and have improved nutritional value, as compared to isolates obtained from conventional plant protein extracts. Furthermore, it has now been surprisingly found that the plant-protein isolates of the invention are characterized by a capability of whitening food products (e.g., liquid products, such as plant based “milk”, etc.), similar to commercial inorganic food whitening agents such as TiC) . To this end, the plant-protein isolates of the invention can be utilized as a protein source for manufacturing of food product, and / or as a whitening agent providing a healthy alternative for toxic nanoparticulate TiCh.

[0040] According to a first aspect, there is provided a plant protein composition comprising a first protein fraction characterized by a molecular weight (Mw) between 5 kDa and 17 kDa, and a second protein fraction characterized by a Mw between 85 kDa and 105 kDa, wherein:I. a weight portion of the second protein fraction relative to the total plant protein content of the plant protein composition is at least 1 % ;II. a protein content of the plant protein composition is at least 75%; and,III. the plant protein composition is characterized by a total color difference (AE) in CIELAB coordinates of less than 4, relative to titanium dioxide (TiCh) powder.

[0041] As used herein, the term “plant protein” including any grammatical form thereof encompasses any protein that is naturally present or is derived from a plant or a plant part, including a root, a stem, a leaf, a flower, a fruit, and a seed. Each possibility represents a separate embodiment. In some embodiments, the plant protein is derived from a plant. In some embodiments, the plant protein has an amino acid sequence identical to the amino acid sequence of a natural plant protein. One skilled in the art will appreciate that the plant protein may further comprise modifications, such as glycosylation, phosphorylation, methylation,etc.; and / or the plant protein may be covalently conjugated or physically complexed to a polysaccharide.

[0042] In some embodiments, the plant protein composition is a plant protein enriched composition, i.e., any composition that is enriched by plant proteins, as disclosed herein. In some embodiments, the plant protein composition is a plant extract. In some embodiments, the plant protein composition is a plant isolate. In some embodiments, the plant protein composition is a plant isolate derived from a plant extract. In some embodiments, the plant protein composition comprises or consists essentially of synthetic plant proteins. In some embodiments, the plant protein composition comprises or consists essentially of recombinant plant proteins expressed by an expression system (i.e., a microorganism such as bacterium, yeast, etc., or any other organism such as a plant). In some embodiments, the extract or isolate is a liquid. In some embodiments, the extract or isolate is a solid. In some embodiments, the plant protein composition is a dried extract or isolate. In some embodiments, the plant protein composition is a powderous composition, i.e., in the form of a powder.

[0043] In some embodiments, a protein content of the plant protein composition disclosed herein is at least 75%. In some embodiments, the protein content of the plant protein composition is at least 76%, at least 77%, at least 78%, at least 79%, or at least 80%. Each possibility represents a separate embodiment of the invention. In some embodiments, a protein content of the plant protein composition disclosed herein is at least 80%. In some embodiments, the protein content of the plant protein composition is 80-100%, 81-100%, 82-100%, 80-95%, 81-95%, 82-95%, 80-90%, 81-90%, 82-90%, 80-85%, 81-85%, 82-85%, 80-84%, 81-84%, or 82-84%, including any range between. Each possibility represents a separate embodiment of the present invention. In some embodiments, a protein content of the plant protein composition disclosed herein is about 82%. The term “protein content” as used herein encompasses any chemical modification of the plant protein, including any salt or any solvate thereof.

[0044] In some embodiments, a weight portion of the total albumin content (also referred to herein as “total albumin”) relative to the total protein content of the plant protein composition is at least about 20%, at least about 30%, or at least about 40%. Each possibilityrepresents a separate embodiment of the invention. In some embodiments, a weight portion of the total albumin relative to the total proteins of the plant protein composition is at least about 45%, at least about 46%, at least about 47%, at least about 48%, at least about 49%, or at least about 50%. Each possibility represents a separate embodiment of the invention. In some embodiments, the total albumin content (w / w) relative to the total proteins of the plant protein composition is between about 20% and about 100%, between about 25% and about 100%, between about 30% and about 100%, between about 35% and about 100%, between about 40% and about 100%, between about 45% and about 100%, between about 50% and about 100%, between about 20% and about 90%, between about 25% and about 90%, between about 30% and about 90%, between about 35% and about 90%, between about 40% and about 90%, between about 45% and about 90%, between about 50% and about 90%, between about 20% and about 80%, between about 25% and about 80%, between about 30% and about 80%, between about 35% and about 80%, between about 40% and about 80%, between about 45% and about 80%, between about 50% and about 80%, between about 20% and about 70%, between about 25% and about 70%, between about 30% and about 70%, between about 35% and about 70%, between about 40% and about 70%, between about 45% and about 70%, between about 50% and about 70%, between about 20% and about 60%, between about 25% and about 60%, between about 30% and about 60%, between about 35% and about 60%, between about 40% and about 60%, between about 45% and about 60%, between about 50% and about 60%, between about 20% and about 55%, between about 25% and about 55%, between about 30% and about 55%, between about 35% and about 55%, between about 40% and about 55%, between about 45% and about 55%, or between about 50% and about 55%. Each possibility represents a separate embodiment of the present invention.

[0045] As used herein, the term “albumin” encompasses plant proteins that are soluble in water as defined by Osborne classification method. It is mentioned that the term “albumin” as used herein is not limited to a specific type of proteins, but rather encompasses a family of proteins, herein termed “albumin proteins” (or “albumin species”). Exemplary albumins include, but are not limited to, pea albumins (e.g., PAI and PA2), lectin (~17 kDa), protease inhibitors, and lipoxygenase (97-100kDa).

[0046] The albumin disclosed herein is substantially devoid of a globulin protein. As used herein, the term “globulin” refers to proteins which are soluble in dilute salt solutions. In some embodiments, albumin comprises a mixture of seed-derived proteins. In some embodiments, albumin comprises seed storage protein(s).

[0047] In some embodiments, any one of the protein fractions disclosed herein comprises a plurality of (e.g., at least 2) proteins. In some embodiments, any one of the protein fractions disclosed herein comprises at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, or at least 9 proteins. Each possibility represents a separate embodiment. In some embodiments, any one of the protein fractions disclosed herein comprises 2-100, 2-75, 2-50, 2-25, 2-20, 2- 15, or 2-10 proteins, including any integer between. Each possibility represents a separate embodiment of the invention.

[0048] In some embodiments, a first protein fraction is characterized by a Mw between about 5 kDa and about 17 kDa, including each value within the specified range. In some embodiments, a first protein fraction is characterized by a Mw between 5 kDa and 16 kDa, between 5kDa and 15 kDa, or between 5 kDa and 14 kDa. Each possibility represents a separate embodiment of the invention.

[0049] In some embodiments, the first protein fraction comprises a plurality of (e.g., at least 2) proteins. In some embodiments, the first protein fraction comprises at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, or at least 9 proteins. Each possibility represents a separate embodiment. In some embodiments, the first protein fraction comprises 2-100, 2- 75, 2-50, 2-25, 2-20, 2-15, or 2-10 proteins, including any integer between. Each possibility represents a separate embodiment of the invention.

[0050] In some embodiments, the first protein fraction disclosed herein comprises at least one protein selected from: (i) a protein having a Mw between about 5 kDa and about 6 kDa, (ii) a protein having a Mw between about 9 kDa and about 11 kDa, (iii) a protein having a Mw between about 13 kDa and about 14 kDa, and (iv) any combination thereof. In some embodiments, the first protein fraction disclosed herein comprises at least one protein having a Mw between about 5 kDa and about 6 kDa, at least one protein having a Mw between about 9 kDa and about 11 kDa, and at least one protein having a Mw between about 13 kDa and about 14 KDa. In some embodiments, at least one protein having a Mw between about5 kDa and about 6 kDa is two proteins. In some embodiments, at least one protein having a Mw between about 8 kDa and about 11 kDa is three proteins. In some embodiments, at least one protein having a Mw between about 13 kDa and about 14 kDa is four proteins. In some embodiments, the first protein fraction comprises two proteins having a Mw between about 5 kDa and about 6 kDa, three proteins having a Mw between about 9 kDa and about 11 kDa, and four proteins having a Mw between about 13 kDa and about 14 kDa. In some embodiments, the first protein fraction consists essentially of two proteins having a Mw between about 5 kDa and about 6 kDa, three proteins having a Mw between about 9 kDa and about 11 kDa, and four proteins having a Mw between about 13 kDa and about 14 kDa.

[0051] In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition is at least 30%. In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition is at least 40%. In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition is at least 45%, at least 50%, or at least 55%. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition is 30-99%, 35-99%, 40-99%, 45-99%, 50-99%, 55-99%, 30-95%, 35-95%, 40-95%, 45-95%, 55-90%, 55-95%, 30-90%, 35-90%,40-90%, 45-90%, 50-90%, 55-90%, 30-80%, 35-80%, 40-80%, 45-80%, 50-80%, 55-80%,30-70%, 35-70%, 40-70%, 45-70%, 50-70%, 55-70%, 30-65%, 35-65%, 40-65%, 45-65%,50-65%, 55-65%, 30-60%, 35-60%, 40-60%, 45-60%, 50-60%, or 55-60%. Each possibility represents a separate embodiment of the invention.

[0052] In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition does not exceed 70%. In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition does not exceed 65%, 64%, 63%, 62%, 61%, 60%, 59%, 58%, 57%, 56%, 50%, 45% or 40% of the proteins in the plant protein composition. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins of the plant protein composition is 30-65%, 35-65%, 40-65%, 40-60%, 40-55%, 40-50%, 50-60%, 50- 64%, 50-63%, 50-62%, 50-61%, 50-60%, 50-59%, 50-58%, 50-57%, 50-56%, 51-64%, 51-63%, 51-62%, 51-61%, 51-60%, 51-59%, 51-58%, 51-57%, 51-56%, 52-63%, 52-62%, 52- 61%, 52-60%, 52-59%, 52-58%, 52-57%, 52-56%, 53-63%, 53-62%, 53-61%, 53-60%, 53- 59%, 53-58%, 53-57%, 53-56%, 54-64%, 54-63%, 54-62%, 54-61%, 54-60%, 54-59%, 54- 58%, 54-57%, 54-56%, 55-64%, 55-63%, 55-62%, 55-61%, 55-60%, 55-59%, 55-58%, 55- 57%, or 55-56% of the proteins in the plant protein composition. Each possibility represents a separate embodiment of the invention.

[0053] In some embodiments, the total albumin content of the first protein fraction (w / w) is at least about 30%. In some embodiments, the total albumin content of the first protein fraction (w / w) is at least about 40%, or at least about 50%. Each possibility represents a separate embodiment of the invention. In some embodiments, the total albumin content of the first protein fraction (w / w) is between 30% and 100%, between 35% and 100%, between 40% and 100%, between 45% and 100%, between 50% and 100%, between 55% and 100%, between 60% and 100%, between 30% and 90%, between 35% and 90%, between 40% and 90%, between 45% and 90%, between 50% and 90%, between 55% and 90%, between 60% and 90%, between 30% and 80%, between 35% and 80%, between 40% and 80%, between 45% and 80%, between 50% and 80%, between 55% and 80%, between 60% and 80%, between 30% and 70%, between 35% and 70%, between 40% and 70%, between 45% and 70%, between 50% and 70%, between 55% and 70%, or between 60% and 70%. Each possibility represents a separate embodiment of the invention.

[0054] In some embodiments, the second protein fraction is characterized by a Mw between about 85 kDa and about 105 kDa, including each value within the specified range. In some embodiments, the second protein fraction is characterized by a Mw between 90 kDa and 105 kDa, between 95 kDa and 105 kDa, between 95 kDa and 100 kDa, between 96 kDa and 100 kDa, between 97 kDa and 100 kDa, between 95 kDa and 99 kDa, between 95 kDa and 98 kDa, between 96 kDa and 99 kDa, between 96 kDa and 98 kDa, between 97 kDa and 99 kDa, or between 97 kDa and 98 kDa. Each possibility represents a separate embodiment of the invention.

[0055] In some embodiments, the second protein fraction comprises at least 2 proteins. In some embodiments, the second protein fraction comprises 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4,1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, or 2-3 proteins. Each possibility represents a separate embodiment of the invention.

[0056] In some embodiments, the second protein fraction disclosed herein comprises at least one protein having a Mw between about 95 kDa and about 100 kDa. In some embodiments, at least one protein is two proteins. In some embodiments, the second protein fraction disclosed herein comprises two proteins having a Mw between about 95 kDa and about 100 kDa. In some embodiments, the second protein fraction disclosed herein consists essentially of two proteins having a Mw between about 95 kDa and about 100 kDa.

[0057] A skilled artisan will appreciate that each protein species includes a plurality of protein molecules having distinct isotope distribution, and as a consequence a different molecular weight. Accordingly, the term “Mw” as used herein, refers to an average molecular weight of a protein, as determined by mass spectrometry (MS, LC-MS, GC-MS, MALDI, etc.) or by methods such as, e.g., SDS-PAGE.

[0058] In some embodiments, a weight portion of the second protein fraction relative to the total plant proteins of the plant protein composition is at least 1%. In some embodiments, a weight portion of the second protein fraction is at least 1%, at least 2%, at least 3%, at least 5%, at least 10%, at least 12.5%, at least 15%, at least 20%, at least 25%, or at least 30%. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of the second protein fraction relative to the total plant proteins of the plant protein composition is between 1-50%, 1-40%, 1-30%, 1-25%, 1-20%, 1-15%, 1-10%, or 1- 5%, including any range between. Each possibility represents a separate embodiment of the invention.

[0059] In some embodiments, the second protein fraction comprises a lipoxygenase. In some embodiments, the lipoxygenase is inactivated.

[0060] In some embodiments, the plant protein composition comprises a third protein fraction characterized by a Mw between about 23 kDa and about 27 kDa, or between 23.5 and 26.5 kDa, including each value within the specified ranges.

[0061] In some embodiments, the third protein fraction disclosed herein comprises at least one protein selected from: (i) a protein having a Mw between about 23 kDa and about 24kDa, (ii) a protein having a Mw between about 26 kDa and about 27 kDa, and (iii) any combination thereof. In some embodiments, the third protein fraction disclosed herein comprises at least one protein having a Mw between about 23 kDa and about 24 kDa, and at least one protein having a Mw between about 26 kDa and about 27 kDa. In some embodiments, at least one protein having a Mw between about 23 kDa and about 24 kDa is one protein. In some embodiments, at least one protein having a Mw between about 26 kDa and about 27 kDa is one protein. In some embodiments, the third protein fraction consists essentially of one protein having a Mw between 23 kDa and 24 kDa, and one protein having a Mw between 26 kDa and 27 kDa.

[0062] In some embodiments, a weight portion of the third protein fraction relative to the total plant proteins of the plant protein composition is at least 5%, or at least 10%. In some embodiments, a weight portion of the third protein fraction relative to the total protein content in the plant protein composition is at least 11%, at least 12%, at least 13%, at least 14%, at least 15%, at least 16%, at least 17 %, at least 18%, or at least 19%. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of the third protein fraction relative to the total plant proteins of the plant protein composition is 5-40%, 10-40%, 15-40%, 5-35%, 10-35%, 15-35%, 5-30%, 10-30%, 15-30%, 5-25%, 10- 25%, 15-25%, 5-20%, 10-20%, or 15-20%. Each possibility represents a separate embodiment of the invention.

[0063] In some embodiments, the total albumin content of the third protein fraction (w / w) is at least about 80%. In some embodiments, the total albumin content of the third protein fraction (w / w) is 80-100%, 85-100%, or 88-100%. Each possibility represents a separate embodiment of the invention.

[0064] In some embodiments, the plant protein composition comprises a fourth protein fraction characterized by a Mw between about 27 kDa and about 35 kDa, including each value within the specified range. In some embodiments, the fourth protein fraction is characterized by a Mw between 27.1 kDa and 31 kDa, including each value within the specified range.

[0065] In some embodiments, the fourth protein fraction disclosed herein comprises at least one protein selected from: (i) a protein having a Mw between about 27 kDa and about28 kDa, (ii) a protein having a Mw between about 30 kDa and about 31 kDa, and (iii) any combination thereof. In some embodiments, the fourth protein fraction disclosed herein comprises at least one protein having a Mw between about 27 kDa and about 28 kDa, and at least one protein having a Mw between about 30 kDa and about 31 kDa. In some embodiments, at least one protein having a Mw between about 27 kDa and about 28 kDa is one protein. In some embodiments, at least one protein having a Mw between about 30 kDa and about 31 kDa is one protein. In some embodiments, the fourth protein fraction consists essentially of one protein having a Mw between 27 kDa and 28 kDa, and one protein having a Mw between 30 kDa and 31 kDa.

[0066] In some embodiments, a weight portion of the fourth protein fraction relative to the total plant protein content of the plant protein composition is at least 4%. In some embodiments, a weight portion of the fourth protein fraction relative to the total plant protein content of the plant protein composition is at least 5%, at least 6%, at least 7%, or at least 8%. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of the fourth protein fraction relative to the total plant proteins of the plant protein composition is 1-50%, 1-40%, 1-30%, 1-20%, 4-20%, 5-20%, 6-20%, 7-20%, 8-20%, 4-15%, 5-15%, 6-15%, 7-15%, 8-15%, 4-10%, 5-10%, 6-10%, 7- 10%, 8-10%, or 8-9%. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of the fourth protein fraction relative to the total plant proteins of the plant protein composition is about 8%.

[0067] In some embodiments, the plant protein composition comprises a fifth protein fraction characterized by a Mw between about 35 kDa and about 70 kDa, including each value within the specified range. In some embodiments, the fifth protein fraction is characterized by a Mw between 35 kDa and 65 kDa, including each value within the specified range.

[0068] In some embodiments, the fifth protein fraction disclosed herein comprises at least one protein selected from: (i) a protein having a Mw between about 40 kDa and about 55 kDa, (ii) a protein having a Mw between about 58 kDa and about 62 kDa, and (iii) any combination thereof. In some embodiments, the fifth protein fraction disclosed herein comprises at least one protein having a Mw between about 58 kDa and about 62 kDa. Insome embodiments, at least one protein having a Mw between about 58 kDa and about 62 kDa is one protein.

[0069] In some embodiments, a weight portion of the fifth protein fraction relative to the total plant proteins of the plant protein composition is at least 0.5%. In some embodiments, a weight portion of a protein having a Mw between about 58 kDa and about 62 kDa, relative to the total plant proteins of the plant protein composition is at least 0.5%. In some embodiments, a weight portion of a protein having a Mw between about 58 kDa and about 62 kDa relative to the total plant proteins of the plant protein composition is 1-50%, 1-40%, 1-30%, 1-20%, 1-10%, 1-5%, 0.5-50%, 0.5-40%, 0.5-30%, 0.5-20%, 0.5-10%, or 0.5-5%, including any range between. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight portion of a protein having a Mw between about 58 kDa and about 62 kDa, relative to the total plant protein content of the plant protein composition is about 0.8%.

[0070] In some embodiments, the plant protein composition is characterized by a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction of 2.25 or less, for example, 0.01-2.25, 0.02-2.2, or 0.02-2.17, including each value within the specified ranges. In some embodiments, the ratio of a reference protein fraction (having a molecular weight between 40 kDa and 70 kDa) to the second protein fraction is up to 1.9, 1.8, 1.7, 1.6, or 1.5. Each possibility represents a separate embodiment.

[0071] In some embodiments, the plant protein composition is characterized by a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction of 2.2 or less. In some embodiments, the plant protein composition is characterized by a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction of 2.1 or less. In some embodiments, the plant protein composition is characterized by a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction of 2.0 or less. In some embodiments, the plant protein composition is characterized by a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction of1.75 or less. In some embodiments, the plant protein composition is characterized by a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction of 1.5 or less.

[0072] In some embodiments, a weight portion of the reference protein fraction relative to the total plant proteins of the plant protein composition is at least 0.5%. In some embodiments, a weight portion of the reference protein fraction relative to the total plant proteins of the plant protein composition is 1-50%, 1-40%, 1-30%, 1-20%, 1-10%, 1-5%, 0.5-50%, 0.5-40%, 0.5-30%, 0.5-20%, 0.5-10%, 0.5-5%, 0.5-3%, or 0.5-1%, including any range between. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein composition is devoid of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa.

[0073] In some embodiments, the plant protein composition is characterized by a total color difference ( AE) in CIELAB coordinates of less than 4, relative to a reference white. In some embodiments, a reference white comprises titanium dioxide (TiC ) powder. In some embodiments, a reference white is the CIELAB coordinates of TiO powder. In some embodiments, the plant protein composition (e.g., a powderous composition) is characterized by a total color difference (AE) in CIELAB coordinates of less than 5, less than 4, less than 3, less than 2, relative to TiO powder (or reference white). Each possibility represents a separate embodiment. In some embodiments, AE in CIELAB coordinates between the plant protein composition disclosed herein and titanium dioxide (TiO ) powder is between 0.1 and 4, between 0.1 and 3.5, between 0.1 and 3, between 0.1 and 2.5, or between 0.1 and 2, including any range between. In some embodiments, the plant protein composition is a liquid and is characterized AE in CIELAB coordinates of below 3, below 2, below 1, or below 0.5, relative to TiCh powder (or reference white).

[0074] As used herein, the term “CIELAB coordinates” refers to three-dimensional coordinates used for color measurement in a CIELAB color space. The CIELAB color space, also referred to as L*a*b*, is a color space defined by the International Commission on Illumination (abbreviated CIE).

[0075] The term “reference white”, “target white”, and “gold standard white” are herein used interchangeably and refer to a set of tristimulus values or chromaticity coordinates thatserve to define a color as “white”. In some embodiments, white is an achromatic white. As used herein, “an achromatic white” is a white color in which the red, green, and blue codes are equal. In some embodiments, a reference white comprises CIELAB color coordinates corresponding to: L*: 94-100, a*: -0.1 to 0.1, and b*: -0.1 to 5.

[0076] “Delta E” (“dE” or “AE”) levels, used herein, refer to the difference between an examined color and a reference color or a gold standard color. Lower delta E values indicate greater accuracy, while high delta E levels indicate a significant mismatch between the examined color and the reference color. Delta E is measured on a scale of from 0 to 100, where 0 indicates color similarity / identity, and 100 indicates complete distortion. In some specific embodiments, dE value < 2.75 is not perceptible by the human eye or perceptible through a close observation.

[0077] In some embodiments, the plant protein composition is characterized by: L* CIELAB value in a range between about 90 and about 95, including each value within the specified range. In some embodiments, the plant protein composition is characterized by a* CIELAB value in a range between about (-0.1) and about 0.4, including each value within the specified range. In some embodiments, the plant protein composition is characterized by b* CIELAB value in a range between about 4 and about 10, including each value within the specified range. In some embodiments, the plant protein composition is characterized by: L* CIELAB value in a range between about 90 and about 95, by a* CIELAB value in a range between about (-0.1) and about 0.4, and by b* CIELAB value in a range between about 4 and about 10, including each value within the specified ranges. In some embodiments, L* CIELAB value of the plant protein composition is between 90 and 94, including each value within the specified range. In some embodiments, a* CIELAB value of the plant protein composition is between (-0.1) and 0.4, including each value within the specified range. In some embodiments, b* CIELAB value of the plant protein composition is between 4 and 10, between 5 and 10, between 4 and 9, between 5 and 9, between 4 and 8, or between 5 and 8, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein isolate composition is characterized by L* CIELAB value in a range between about 80 and about 95; a* CIELAB value in a range between about (-2) and about 1.5; and b* CIELAB value in a range between about 3 and about 10.0, including each value within the specified ranges.

[0078] In some embodiments, the plant protein composition is characterized by reflectance to transmittance ratio when measured at 400 nm in a range between about 50% and about 80%, including each value within the specified range. In some embodiments, the plant protein composition is characterized by reflectance to transmittance ratio when measured at 400 nm in a range between about 50% and about 75%, between about 55% and about 80%, between about 55% and about 75%, between about 60% and about 80%, or between about 60% and about 75%, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein composition is characterized by reflectance to transmittance ratio when measured at 400 nm in a range between about 60% and about 75%, including each value within the specified range.

[0079] In some embodiments, the plant protein composition disclosed herein comprises trace amounts of a hydrocolloid. In some embodiments, the plant protein composition comprises trace amounts of hydrocolloid-forming polysaccharides. In some embodiments, the plant protein composition further comprises trace amounts of an alkaline earth metal salt.

[0080] As used herein, the term “trace amounts” refers to an amount within the range of 1 ppm and 0.1 % (w / w), or between 10 ppm and 1% (w / w), between 10 and 10,000 ppm, between 10 and 5,000 ppm, between 10 and 1,000 ppm, between 10 and 500 ppm, or between 10 and 100 ppm by dry weight of the plant protein composition, including any range between. Each possibility represents a separate embodiment. In some embodiments, trace amounts refer to amounts of a hydrocolloid of at most 20,000 ppm, at most 15,000 ppm, at most 10,000 ppm, at most 5,000 ppm, at most 1,000 ppm, at most 500 ppm, at most 250 ppm, or at most 100 ppm. Each possibility represents a separate embodiment.

[0081] In some embodiments, the plant protein composition is a powderous plant protein composition. In some embodiments, the water content of the plant protein composition is below 10% (w / w), for example between 0.1% and 10%, between 0.1% and 9%, between 0.1% and 8%, between 0.1% and 7%, between 0.1% and 6%, between 0.1% and 5%, between 0.1% and 4%, between 0.1% and 3%, between 0.1% and 2%, between 0.1% and 1%, or between 0.1% and 0.5%. Each possibility represents a separate embodiment of the invention.

[0082] In some embodiments, the powderous plant protein composition is characterized by Dv(90) between about 30 pm and about 50 pm. In some embodiments, the powderous plant protein composition is characterized by Dv(90) between about 100 pm and about 150 pm, including each value within the specified range. In some embodiments, the powderous plant protein composition is characterized by Dv(10) between about 1 pm and about 5 pm, including each value within the specified range. In some embodiments, Dv(10) of the powderous plant protein composition is between about 2 pm and about 4 pm, including each value within the specified range. In some embodiments, Dv(10) of the powderous plant protein composition is between about 4 pm and about 10 pm, including each value within the specified range. In some embodiments, the powderous plant protein composition is characterized by Dv(50) between about 10 pm and about 20 pm, including each value within the specified range. In some embodiments, the powderous plant protein composition is characterized by Dv(50) between about 35 pm and about 55 pm, including each value within the specified range. In some embodiments, the powderous plant protein composition is characterized by Dv(90) between about 30 pm and about 50 pm, Dv(10) between about 1 pm and about 5 pm, and Dv(50) between about 10 pm and about 20 pm, including each value within the specified ranges.

[0083] As used herein, the term “Dv” refers to a particle size. Particle size parameters can be determined, for example, by laser diffraction particle size analyzer. The values of Dv(10), Dv(50), and Dv(90), respectively, represent 10%, 50%, and 90% of the product below the named micron size, as determined, for example, by DLS. For example, D(v)90 of about 50 pm means that 90% of the particles in the composition (w / w) have sizes which are below about 50 pm.

[0084] In some embodiments, the powderous plant protein composition is characterized by a specific surface area of at least 300 m2 / kg. In some embodiments, the powderous plant protein composition is characterized by a specific surface area of at least 400 m2 / kg. In some embodiments, the powderous plant protein composition is characterized by a specific surface area of at least 500 m2 / kg. In some embodiments, the powderous plant protein composition is characterized by a specific surface area of at least 600 m2 / kg. In some embodiments, the powderous plant protein composition is characterized by a specific surface area of at least 700 m2 / kg. In some embodiments, the powderous plant protein composition is characterizedby a specific surface area of between about 600 m2 / kg and about 1,400 m2 / kg, between about 600 m2 / kg and about 1,300 m2 / kg, between about 700 m2 / kg and about 1,400 m2 / kg, or between about 700 m2 / kg and about 1,300 m2 / kg, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, the powderous plant protein composition is characterized by a specific surface area of between 700 m2 / kg and 1,250 m2 / kg, including each value within the specified range.

[0085] As used herein, specific surface area (SSA) is a property of solids defined as the total surface area of a material per unit of mass.

[0086] In some embodiments, the powderous plant protein composition is characterized by uniformity of between about 0.6 and about 1.2, including each value within the specified range. In some embodiments, the powderous plant protein composition is characterized by span of between about 2 and about 3.7, including each value within the specified range. In some embodiments, the powderous plant protein composition is characterized by at least one of: D[3,2] between about 4 pm and about 9 pm, D[3,2] of between about 15 pm and about 20 pm; D[4,3] between about 15 pm and about 25 pm, D[4,3] of between about 50 pm and about 65 pm, and combinations thereof. In some embodiments, the particle size values, uniformity, and span are as determined by Light Scattering (e.g., laser diffraction particle size analyzer).

[0087] In some embodiments, the plant protein composition is characterized by a total fat content (w / w) below 9%. In some embodiments, the plant protein composition is characterized by a total fat content (w / w) below 8%, below 7%, below 6%, below 5%, below 4%, below 3%, or below 2%. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein composition is characterized by a total fat content (w / w) of between 0.5% and 9%, between 0.5% and 8%, between 0.5% and 7%, between 0.5% and 6%, between 0.5% and 5%, between 0.5% and 4%, between 0.5% and 3%, between 0.5% and 2%, between 1% and 9%, between 1% and 8%, between 1% and 7%, between 1% and 6%, between 1% and 5%, between 1% and 4%, between 1% and 3%, or between 1% and 2%, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant proteincomposition is characterized by a total fat content (w / w) of between about 0.5% and about 2%, including each value within the specified range.

[0088] In some embodiments, the plant protein composition is characterized by a carbohydrates content (w / w) of at least 0.1%. In some embodiments, the plant protein composition is characterized by a carbohydrates content (w / w) of at least 0.5%, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, or at least 6%. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein composition is characterized by a carbohydrates content (w / w) of between 0.1% and 10%, between 0.5% and 10%, between 1% and 10%, between 1% and 9%, between 1% and 8%, between 2% and 10%, between 2% and 9%, between 2% and 8%, between 3% and 10%, between 3% and 9%, between 3% and 8%, between 4% and 10%, between 4% and 9%, between 4% and 8%, between 5% and 10%, between 5% and 9%, between 5% and 8%, between 6% and 10%, between 6% and 9%, or between 6% and 8%, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein composition is characterized by a carbohydrates content (w / w) of between about 6% and about 8%, including each value within the specified range.

[0089] Methods for determining nutritional content of food plant protein compositions are well known in the art. Non-limiting examples include detection of moisture (or water content) by loss of mass at 102 °C, detection of total protein content by analysis of total nitrogen (e.g., by Dumas or Kjeldahl methods), detection of total fat content by a solvent extraction, or by secondary methods, e.g., by nuclear magnetic resonance spectroscopy (NMR), and detection of carbohydrates content by a liquid chromatography technique, e.g., by high-performance anion exchange (HPAE) chromatography.

[0090] In some embodiments, the plant protein composition disclosed herein is characterized by an essential amino acid (EAA) content greater than about 20% (w / w) relative to the total amino acid content of the plant protein composition. In some embodiments, the plant protein composition disclosed herein is characterized by an essential amino acid (EAA) content greater than about 25% (w / w) relative to the total amino acid content of the plant protein composition. In some embodiments, the plant proteincomposition disclosed herein is characterized by an essential amino acid (EAA) content greater than about 35% (w / w) relative to the total amino acid content of the plant protein composition. In some embodiments, the plant protein composition disclosed herein is characterized by an essential amino acid (EAA) content greater than about 38% (w / w) relative to the total amino acid content of the plant protein composition. In some embodiments, the EAA content of the plant protein composition relative to the total amino acid content (w / w) is at least about 38%, at least about 39%, or at least about 40%. Each possibility represents a separate embodiment of the invention. In some embodiments, the plant protein composition is characterized by an EAA content (w / w) between about 38% and about 50%, between about 39% and about 50%, between about 40% and about 50%, between about 38% and about 45%, between about 39% and about 45%, between about 40% and about 45%, between about 38% and about 44%, between about 39% and about 44%, between about 40% and about 44%, between about 38% and about 43%, between about 39% and about 43%, or between about 40% and about 43%, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, the EAA content of the plant protein composition relative to the total amino acid content (w / w) is between about 40% and about 45%, including each value within the specified range.

[0091] As used herein, the term “essential amino acid EAA” encompasses an amino acid that cannot be produced in a human subject. As used herein, an EAA is selected from: histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine.

[0092] In some embodiments, lysine constitutes at least about 18% (w / w) of the EAA content in the plant protein composition disclosed herein. In some embodiments, lysine constitutes at least about 20% (w / w) of the EAA content. In some embodiments, lysine constitutes between about 18% and about 25%, between about 19% and about 25%, between about 20% and about 25%, between about 18% and about 24%, between 19% and about 24%, or between 20% and about 24% of the EAA content, including each value within the specified ranges. Each possibility represents a separated embodiment of the invention. In some embodiments, lysine constitutes between about 20% and about 24% (w / w) of the EAA content, including each value within the specified range.

[0093] In some embodiments, branched chain amino acids (BCAAs) constitute at least 30% (w / w) of the EAA content. In some embodiments, branched chain amino acids (BCAAs) constitute at least 35% (w / w) of the EAA content. In some embodiments, branched chain amino acids (BCAAs) constitute at least 40% (w / w) of the EAA content. In some embodiments, BCAAs constitute between about 40% and about 60%, between about 45% and about 60%, between about 40% and about 55%, or between about 45% and about 55% (w / w) of the EAA content, including each value within the specified ranges. Each possibility represents a separated embodiment of the invention. In some embodiments, BCAAs constitute between about 45% and about 54%, between about 46% and about 54%, between about 47% and about 54%, between about 48% and about 54%, between about 49% and about 54%, between about 45% and about 53%, between about 46% and about 53%, between about 47% and about 53%, between about 48% and about 53%, between about 49% and about 53%, between about 45% and about 52%, between about 46% and about 52%, between about 47% and about 52%, or between about 48% and about 52% (w / w) of the EAA content, including each value within the specified ranges. Each possibility represents a separated embodiment of the invention. In some embodiments, BCAAs constitute about 50% (w / w) of the EAA content.

[0094] As used herein, the term “branched chain amino acid (BCAA)” refers to an amino acid selected from: leucine, isoleucine, and valine.

[0095] In some embodiments, a weight content of glutamic acid relative to the total plant proteins in the plant protein composition is between about 5% and about 25%, including each value within the specified range. In some embodiments, a weight content of glutamic acid relative to the total plant proteins in the plant protein composition is between about 5% and about 24%, between about 5% and about 23%, between about 5% and about 22%, between about 5% and about 21%, between about 5% and about 20%, between about 5% and about 19%, between about 5% and about 18%, between about 5% and about 17%, between about 5% and about 16%, or between about 5% and about 15%, including each value within the specified ranges. Each possibility represents a separate embodiment of the invention. In some embodiments, a weight content of glutamic acid relative to the total plant proteins in the plant protein composition is below about 17%, below about 16%, below about 15%, below about 14%, or below about 13%. Each possibility represents a separateembodiment of the invention. In some embodiments, a weight content of glutamic acid relative to the total plant proteins in the plant protein composition is between about 6% and about 15%, between about 7% and about 15%, between about 8% and about 15%, between about 9% and about 15%, between about 10% and about 15%, between about 11% and about 15%, between about 6% and about 14%, between about 7% and about 14%, between about 8% and about 14%, between about 9% and about 14%, between about 10% and about 14%, between about 6% and about 13%, between about 7% and about 13%, between about 8% and about 13%, between about 9% and about 13%, between about 10% and about 13%, or between about 11% and about 13%, including each value within the specified ranges. Each possibility represents a separated embodiment of the invention. In some embodiments, a weight content of glutamic acid relative to the total plant proteins in the plant protein composition is between about 11% and about 13%, including each value within the specified range.

[0096] Methods for determining the content of amino acids, comprising EAAs and / or BCAAs are known in the art. Examples for such methods include, but are not limited to, thin-layer chromatography, high-performance liquid chromatography, liquid chromatography-mass spectrometer, gas chromatography-mass spectrometry, capillary electrophoresis, nuclear magnetic resonance, and amino acid analyzer analysis.

[0097] In some embodiments, the plant disclosed herein is from the family Fabaceae. In some embodiments, the plant disclosed herein is a legume. In some embodiments, the plant proteins disclosed herein are legume proteins.

[0098] As used herein, the term “legume” encompasses a plant in the family Fabaceae (also termed Leguminosae), or a portion thereof. In some embodiments, a portion of legume comprises a fruit or a seed of a legume plant. In some embodiments, a legume is selected from a pea, a soybean, a lentil, a bean, a chickpea, a peanut, a lupin, a mesquite, a carob, a tamarind, an alfalfa, a clover, and any combination thereof. Each possibility represents a separate embodiment. In some embodiments, a legume comprises peas selected from green peas and yellow peas. In some embodiments, a legume comprises lentils selected from green lentils, red lentils, brown lentils, and French lentils. In some embodiments, a legume comprises beans selected from black beans, kidney beans, navy beans, pinto beans,cannellini beans, lima beans, garbanzo beans, adzuki beans, soybeans, and any combination thereof.

[0099] In some embodiments, legume proteins are selected from: pea proteins, soy proteins, chickpea proteins, lentil proteins, bean proteins, peanut proteins, or any combination thereof. Each possibility represents a separate embodiment.

[0100] In some embodiments, the plant protein composition is a plant protein isolate. In some embodiments, the plant protein isolate is derived from a plant extract. In some embodiments, the plant protein isolate is derived from a portion of the plant. In some embodiments, the plant protein isolate is derived from a seed. In some embodiments, a seed comprises a legume seed. In some embodiments, a seed is a legume seed. In some embodiments, a plant protein isolate is derived from a seed of a legume plant.

[0101] In some embodiments, a legume plant comprises a pea. In some specific embodiment, a legume plant is a pea (Pisum sativum). In a yet more specific embodiment, a pea is a green pea. In a further specific embodiment, a pea is a yellow pea. In some embodiments, a plant protein comprises 2S albumin. In some embodiments, 2S albumin comprises lectin, albumin 2, albumin 1A, or any combination thereof. Each possibility represents a separate embodiment.Green pea protein composition

[0102] In another aspect, there is provided a protein composition comprising or consisting essentially of plant proteins (e.g., recombinant, synthetic, or natural plant proteins), wherein the protein content of the protein composition is at least 80%, the albumin content of the protein composition is at least 40%; wherein the protein composition comprises at least 50%, or between 50% and 70% of a first protein fraction, a second protein fraction, a third protein fraction, and a fourth protein fraction; wherein the first protein fraction, the second protein fraction, the third protein fraction, and the fourth protein fraction are as disclosed herein. In some embodiments, the protein composition is a plant protein composition. In some embodiments, the protein composition comprises or consist essentially of green pea proteins. In some embodiments, the protein composition is a green pea protein (GPP) composition.

[0103] In some embodiments, a weight portion of the first protein fraction relative to the total plant proteins is at least 50%, wherein the first protein fraction is characterized by at least one of: (i) total albumin content of at least 50% w / w; (ii) comprises at least about 10% w / w of pea albumin protein of late embryogenesis; (iii) at least about 13% of pea albumin- 1A; and (iv) at least about 13% of pea albumin-lE.

[0104] In some embodiments, the weight portion of the first protein fraction relative to the total plant proteins of the GPP composition is about 55%. In some embodiments, the total albumin content within the GPP composition is between 50 and 60% w / w, including each value within the specified range. In some embodiments, the first protein fraction of the GPP composition comprises at least one protein selected from: defensin- 2 (Uniprot No. P81930), seed albumin 2 (Uniprot No. Q5ZGL2), non-specific lipid transfer protein (Uniprot No. AOA182DV18), seed trypsin / chymotrypsin inhibitor IVA (Uniprot No. Q41065), seed albumin protein of late embryogenesis 1 abundant group (Uniprot No. AOA8J9XGW9), albumin-lE (Uniprot No. P62930), albumin-1 A (Uniprot No. P62926), albumin-lF (Uniprot No. P62931), albumin-lD (Uniprot No. P62929), or any combination thereof. Each possibility represents a separate embodiment.

[0105] In some embodiments, the first protein fraction of the GPP composition comprises a plurality of proteins selected from: defensin- 2, seed albumin 2, non-specific lipid transfer protein, seed trypsin / chymotrypsin inhibitor IVA, seed albumin protein of late embryogenesis 1 abundant group, albumin-lE, albumin-lA, albumin-lF, albumin-lD, or any combination thereof. Each possibility represents a separate embodiment.

[0106] In some embodiments, the albumin proteins within the first protein fraction of the GPP composition, comprise at least one protein selected from: fragment of seed albumin 2, seed albumin protein of late embryogenesis 1 abundant group, albumin- IE, albumin- 1 A, albumin- IF, albumin- ID, or any combination thereof. Each possibility represents a separate embodiment.

[0107] In some embodiments, the first protein fraction of the GPP composition consists essentially of: defensin- 2, seed albumin 2, non-specific lipid transfer protein, seed trypsin / chymotrypsin inhibitor IVA, seed albumin protein of late embryogenesis 1 abundantgroup, albumin-lE, albumin-lA, albumin-lF, and albumin-lD. Each possibility represents a separate embodiment.

[0108] In some embodiments, the content (w / w) of seed albumin protein of late embryogenesis 1 abundant group in the first protein fraction of the GPP composition is at least 10%, for example between about 8% and about 14%, between about 8% and about 13%, or between about 8% and about 12%, including each value within the specified ranges.

[0109] In some embodiments, the content (w / w) of albumin 1 A in the first protein fraction of the GPP composition is at least 10%, for example between about 10% and about 16%, between about 10% and about 15%, between about 11% and about 16%, between about 11% and about 15%, between about 12% and about 16%, between about 12% and about 15%, between about 13% and about 16%, or between about 13% and about 15%, including each value within the specified ranges.[001 10] In some embodiments, the content (w / w) of albumin IE in the first protein fraction of the GPP composition is at least 10%, for example between about 10% and about 16%, between about 10% and about 15%, between about 11% and about 16%, between about 11% and about 15%, between about 12% and about 16%, between about 12% and about 15%, between about 13% and about 16%, or between about 13% and about 15%, including each value within the specified ranges.

[0111] In some embodiments, the second protein fraction of the GPP composition comprises a lipoxygenase (EC No. 1.13.11). In some embodiments, the second protein fraction of the GPP composition comprises or consist essentially of seed linoleate 9s- lypoxygenase-2 (Uniprot No. P14856) and seed linoleate 9s-lypoxygenase-3 (P09918). In some embodiments, the weight portion of the second protein fraction relative to the total plant proteins of the GPP composition is about 3-4%.[001 12] In some embodiments, a weight ratio between the linoleate 9s-lipoxygenase-3 and linoleate 9s-lipoxygenase-2 within the second protein fraction of the GPP composition is in a range between about 5:1 and about 1: 1, for example between about 4: 1 and about 1: 1, between about 3: 1 and about 1: 1, between about 5:1 and about 2:1, between about 4: 1 and about 2: 1, or between about 3:1 and about 2:1, including all iterations of ratios within thespecified ranges. In some embodiments, a weight ratio between the linoleate 9s- lipoxygenase-3 and linoleate 9s-lipoxygenase-2 is in a range between about 3: 1 and 2: 1.[001 13] In some embodiments, the third protein fraction of the GPP composition comprises albumin 2 (Uniprot No. PO8688) and dehydrin DH3 (Uniprot No. P28641). In some embodiments, the third protein fraction consists essentially of albumin 2 and dehydrin DH3. In some embodiments, the albumin proteins within the third protein fraction of the GPP composition comprise albumin 2.

[0114] In some embodiments, the weight portion of the third protein fraction relative to the total plant proteins of the GPP composition is about 19%.

[0115] In some embodiments, the fourth protein fraction of the GPP composition comprises dehydrin 2 (Uniprot No. Q04117), lectin (Uniprot No. P02867), or both. In some embodiments, the fourth protein fraction of the GPP composition consists essentially of dehydrin 2 and lectin. In some embodiments, the weight portion of the fourth protein fraction relative to the total plant proteins of the GPP composition is about 8%.[001 16] In some embodiments, the GPP composition further comprises a fifth protein fraction. In some embodiments, the weight portion of the fifth protein fraction relative to the total plant proteins of the GPP composition is about 0.8-1%.

[0117] In some embodiments, the fifth protein fraction of the GPP composition comprises seed biotin-containing protein SBP65 (Uniprot No. Q41060). In some embodiments, the fifth protein fraction of the GPP composition consists essentially of seed biotin-containing protein SBP65.

[0118] In some embodiments, the GPP composition is characterized by an essential amino acid (EAA) content greater than 30% w / w. In some embodiments, the GPP composition is characterized by an essential amino acid (EAA) content greater than 40% w / w, for example between 40% and 60% w / w, relative of the total amino acid content. In some embodiments, the lysine content is about 20% w / w of the EAA content of the GPP composition. In some embodiments, BCAAs constitute about 50% w / w of the EAA content of the GPP composition.[001 19] In some embodiments, a weight content of glutamic acid is between about 11 and about 13%, relative to the total amino acid content of the GPP composition. An exemplary proteins composition / protein abundancy of the GPP composition (e.g., in extract form) is as disclosed in Tables 1 and 2.

[0120] In some embodiments, the plant protein composition of the invention is characterized by similar food whitening properties, as compared to TiO2 powder. TiO2 has long been used as a food coloring agent, to whiten or provide a cloudy effect, being included in food products (e.g., chewing gum, sweets, sauces, and baked goods).

[0121] In some embodiments, the plant protein composition is characterized by improved organoleptic properties, as compared to a plant protein isolate obtained from the same plant.

[0122] As used herein, the term “organoleptic property” refers to a characteristic of food selected from: color, taste, smell, and texture. In some embodiments, the plant protein composition is characterized by improved whitening color as compared to a commercially available protein isolate obtained from the same plant. In some embodiments, the plant protein composition is characterized by an improved smell, or a reduced smell as compared to a commercially available protein isolate obtained from the same plant. In some embodiments, the plant protein composition is characterized by an improved taste or reduced aftertaste as compared to a commercially available protein isolate obtained from the same plant.Methods of preparation

[0123] As opposed to hitherto reported plant protein isolate compositions produced using aqueous fractionation methods comprising solubilization at alkaline pH followed by a precipitation step of globulin proteins at acidic pH, plant protein isolates in accordance with the invention are advantageously produced by a process that does not require pH modulation and does not include a step of pH-mediated globulin precipitation. In some embodiments, the protein source (e.g., pea flour) used for the preparing of plant protein isolates in accordance with the invention is solubilized in an aqueous medium in a substantially neutral pH (e.g., 6.5-7.5) which is substantially maintained during the preparation process.

[0124] Further, as disclosed and exemplified herein, plant protein isolates in accordance with embodiments of the invention are advantageously produced by a process comprising hydrocolloid-mediated globulin separation. For example, without limitation, a hydrocolloidforming polysaccharide (such as pectin, carrageenan, gelatin, gellan, hyaluronic acid, chitosan, a cellulose, e.g., carboxymethyl cellulose (CMC), a gum, e.g., locust bean gum, xanthan gum, Konjac gum, gellan gum, gum Arabic, and the like, alginate, agar or starch) may be mixed with the aqueous extract to substantially encapsulate globulin proteins and water-insoluble plant matter, such that the aggregated encapsulate may be subsequently separated from the aqueous extract. Consequently, an albumin-rich plant extract (which is substantially devoid of globulins) is obtained. In some embodiments, the composition comprises trace amounts of a hydrocolloid. In some embodiments, the plant protein isolate is substantially devoid of a residual hydrocolloid.

[0125] In addition, a protein extract produced by a process as described hereinabove (comprising solubilization in aqueous medium at a neutral pH and hydrocolloid-mediated globulin separation) may be further processed to produce a plant protein isolate of the invention, e.g., by a method selected from salting out (e.g., by addition of saturated ammonium sulfate solution), ethanol -based coagulation, and / or by thermal treatment. In some embodiments, the process comprises a step of heat treatment, in which the extract is heated under conditions sufficient to induce protein denaturation or partial protein denaturation. For example, a thermal treatment used in the preparation of a protein isolate may comprise heating the extract at a temperature of up to 95 °C, e.g., 80-95 °C, including each value within the specified range, for up to 10 minutes.

[0126] It is to be understood, that both the plant protein isolates of the invention and their precursors, i.e., plant protein extracts used in their preparation, following hydrocolloid- mediated globulin separation and prior to the further processing steps described above, such as those disclosed in Tables 1 and 2, are encompassed within the scope of the invention.

[0127] Finally, the process may further comprise dehydration of the resulting protein isolate, for example using spray drying, or lyophilization or other common drying methods to obtain a substantially dry composition (e.g., a powderous composition). For example, theresulting precipitate may be centrifuged and dried to obtain a white powder having a Loss on Drying (LOD) of 20% or less, preferably 10% or less.

[0128] It is to be understood that the plant protein isolates of the invention need not be used under conditions amenable with enzymatic activity in order to be used as a whitening agent in food products. In fact, the plant protein isolates of the invention are typically substantially devoid of enzymatic activity, for example due to inclusion of production steps such as thermal treatment. Thus, the plant protein isolates of the invention may conveniently also be used as whitening agents in manufacture processes of food products performed under conditions (e.g., temperature, pH) that would attenuate or prevent enzyme-catalyzed reactions (for example, cooking, freezing etc.).

[0129] According to the principles of the present invention, the process for preparing the plant protein isolates of the invention does not include the use of proteases. In accordance with these embodiments, the process is performed without any additional processing which may result in the hydrolytic breakdown (i.e., chemical hydrolysis or enzymatic hydrolysis) of the proteins of the extract or isolate into low molecular weight peptides, thereby altering the ratio between different protein fractions. Enzymatic hydrolysis, also known as the enzymatic breakdown of proteins, is typically performed in the presence of enzymes, i.e., proteases. Proteases are enzymes that catalyze the hydrolysis of peptide bonds, thereby controlling protein degradation. Thus, encompassed by the present invention are plant protein extracts and isolates that are non-proteolytically hydrolyzed.

[0130] Plant protein isolates in accordance with embodiments of the invention are further characterized in that they do not substantially dissolve in water, but rather form a suspension when added to aqueous media. In particular, plant protein isolates in powder form produced by a process comprising thermal treatment and dehydration as described above, are characterized in that they are suspensible (rather than dissolvable) in water under ambient conditions. In some embodiments, there is provided a suspension formed by dispersing a plant protein isolate of the invention (e.g., an isolate in powder form as described herein) in aqueous media.

[0131] In addition, the plant protein isolates of the invention are typically substantially devoid of plant globulin proteins (such as legumin, vicilin and convicilin). In someembodiments, provided are compositions characterized by a globulin content of less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 2.5%, less than 1.25%, less than 1%, less than 0.75%, less than 0.5%, less than 0.25% or less than 0.1% of plant globulin proteins.

[0132] In some embodiments, the plant protein isolates of the invention are characterized by a protein to fat ratio (w / w) of at least 20 and typically at least 30, at least 40, at least 50, at least 55 or at least 58 (e.g., a weight ratio of about 60).Edible compositions

[0133] In some embodiments, the plant protein isolates in accordance with the invention are incorporated with additional ingredients and supplements into edible compositions for use in the food industry.

[0134] In some embodiments, there is provided an edible composition comprising a plant protein isolate in accordance with the invention. In some embodiments, the edible composition comprises at least 0.1% w / w of said plant protein isolate.

[0135] In some embodiments, there is provided an emulsion comprising a plant protein isolate of the invention. In some embodiments, the emulsion is an oil-in-water emulsion. In some embodiments, the emulsion is a water-in-oil emulsion. In some embodiments, emulsions comprising plant protein isolates of the invention may be used in the production of various food products including, but not limited to, dairy product substitutes and condiments (e.g., ketchup and mayonnaise), as will be described in greater detail below.

[0136] In some embodiments, the emulsion comprises 0.03-3% w / w of the plant protein isolate; 20%-98% w / w of an oily phase; and 2-80% w / w of an aqueous phase, including each value within the specified ranges. In some embodiments, there is provided an edible composition comprising: 0.03-3% w / w the plant protein isolate; 20%-98% w / w of an oily phase; and 2-80% w / w of an aqueous phase, including each value within the specified ranges. In some embodiments, the edible composition is in the form of an oil-in-water emulsion comprising 0.03-3% w / w of the plant protein isolate; 20%-98% w / w of an oily phase; and 2-80% w / w of an aqueous phase, including each value within the specified ranges.

[0137] In some embodiments, the emulsion or composition comprises 0.05-2.5% w / w of the plant protein isolate, including each value within the specified range. In some embodiments, the emulsion or composition comprises 0.1-2% w / w of the plant protein isolate, including each value within the specified range. In some embodiments, the emulsion or composition comprises 0.3-1% w / w of the plant protein isolate, including each value within the specified range.

[0138] In some embodiments, the emulsion or composition comprises 30-90% w / w of an oily phase, including each value within the specified range. In some embodiments, the emulsion or composition comprises 40-80% w / w of an oily phase, including each value within the specified range. In some embodiments, the emulsion or composition comprises 45-75% w / w of an oily phase, including each value within the specified range.

[0139] In some embodiments, the oily phase comprises a food-grade oil. In some embodiments, the oil is selected from olive oil, vegetable oil, cottonseed oil, almond oil, canola oil, coconut oil, corn oil, grape seed oil, peanut oil, saffron oil, sunflower oil, rice oil, sesame oil, soybean oil, and combinations thereof. Each possibility represents a separate embodiment.

[0140] In some embodiments, the emulsion or composition comprises 5-70% w / w of an aqueous phase, including each value within the specified range. In some embodiments, the emulsion or composition comprises 10-60% w / w of an aqueous phase, including each value within the specified range. In some embodiments, the emulsion or composition comprises 15-55% w / w of an aqueous phase, including each value within the specified range.

[0141] In some embodiments, the emulsion or composition is substantially devoid of gelforming agents and / or emulsifying agents. In some embodiments, the compositions, emulsions and food products in accordance with the invention are substantially devoid of titanium dioxide.

[0012] In some embodiments, the composition is in the form of an oil-in-water emulsion. In some embodiments, the composition is in the form of an oil-in-water emulgel.

[0143] In some embodiments, the emulsions as disclosed herein comprising the plant protein isolates of the invention may advantageously be used as whitening agents in foodproducts (e.g., milk substitutes). In some embodiments, it is envisioned that compositions comprising plant protein isolates in accordance with the invention used as food additives may provide additional properties to the food product in which they are incorporated, other than their identified use as whitening agents. For example, the compositions may additionally be used as fat substitutes, stabilizers, and / or emulsifiers. Each possibility represents a separate embodiment of the invention.Food products

[0144] In some embodiments, there is provided a food product or a food article comprising the plant protein composition disclosed herein. In some embodiments, the weight content of the plant protein composition disclosed herein in the food product is between about 0.1% and about 30%, between about 0.1% and about 25%, between about 0.1% and about 20%, between about 0.1% and about 15%, between about 5% and about 30%, between about 5% and about 25%, between about 5% and about 20%, or between about 5% and about 15%, including each value within the specified ranges.

[0145] In some embodiments, the food product comprises the plant protein composition disclosed herein in a w / w ratio of between 0.1% and 10%, between 0.1% and 5%, between 0.1% and 3%, between 0.1% and 1%, between 0.5% and 10%, between 0.5% and 5%, between 0.5% and 3%, or between 0.5% and 1%, including each value within the specified ranges.

[0146] In some embodiments, the food product comprises the edible composition (e.g., the emulsion composition) disclosed herein. In some embodiments, the weight content of the edible composition disclosed herein in the food product is between about 0.1% and about 30%, between about 0.1% and about 25%, between about 0.1% and about 20%, between about 0.1% and about 15%, between about 5% and about 30%, between about 5% and about 25%, between about 5% and about 20%, or between about 5% and about 15%, including each value within the specified ranges.

[0147] According to some embodiments, the food product refers to a non-dairy product. As used herein, the term “non-dairy” includes all types of products that are substantially devoid of milk and milk products that are derived from mammalian source. According to some embodiments, a food product refers to a dairy product.

[0148] In some embodiments, the food product is a liquid. According to some embodiments, the liquid is a beverage and / or a beverage product. As used herein, the term “beverage” refers to a substantially aqueous drinkable composition suitable for human consumption. Non-limiting examples of beverages include water, soft drinks, juices based on fruit extracts, juices based on vegetable extracts, plant milk (e.g., soy milk, almond milk, rice milk, coconut milk etc.), artificial or alternative milk, artificial or alternative yoghurt, and any combination thereof. Each possibility represents a separate embodiment.

[0149] In some embodiments, the food product is selected from meat substitute, fat substitute, dairy substitute, plant-based products, fish substitute, egg substitute, or any combination thereof. Each possibility represents a separate embodiment. In some embodiments, the food product is a vegetarian food product. In some embodiments, the food is a vegan food product. In some embodiments, the food product is devoid of an animal matter.

[0150] In some embodiments, the food product is a protein enriched food product. In some embodiments, the food product is a food grade product. As used herein, the term “food grade” refers to a product consisting essentially of food-grade ingredients approved for human consumption by a corresponding regulatory authority (i.e., GRAS). The concentration of each of the constituents within the food grade product are such that it does not exceed a toxicity limit for the specific constituent as determined by the corresponding regulatory authority.

[0151] According to the principles of the present invention, the plant protein composition is useful as a whitening agent. In some embodiments, the food product comprising the plant protein composition disclosed herein has a white color. In some embodiments, the food product comprises the plant protein composition disclosed herein at an amount (e.g., between 0.1 and 10% w / w) sufficient for obtaining a food product characterized by a total color difference (AE) in CIELAB coordinates of less than 15, less than 10, less than 8, less than 4, or less than 3, relative to a reference white, as disclosed herein.

[0152] In some embodiments, there is provided a food product comprising the emulsion or composition disclosed herein. According to various embodiments, the food product is selected from bakery products, dairy products, dairy substitute products, confectionaries,meat or poultry products, meat or poultry substitute products, fish products, fish substitute products, snack foods, and sauces. Each possibility represents a separate embodiment. In some embodiments, the food product comprises bakery products selected from bread, bread sticks, puff pastry, choux pastry, croissant, muffin, biscuit, pastries, bourekas, brioche, pizza, tortillas, pound cake, and cookies. Each possibility represents a separate embodiment. In some embodiments, the food product comprises dairy products selected from milk, cheese, yogurt, pudding, mousse, desert, cream, and ice cream. Each possibility represents a separate embodiment. In some embodiments, the food product comprises dairy substitute products selected from soy / rice / almond / oats-based milk, cheese, yogurt, pudding, mousse, desert, cream, and ice cream. Each possibility represents a separate embodiment. In some embodiments, the food product comprises confectionaries selected from chocolates, protein bars, pudding, and candies. Each possibility represents a separate embodiment. In some embodiments, the food product comprises meat or poultry products selected from hamburgers, meatballs, sausages, meatloaf, kebab, and nuggets. Each possibility represents a separate embodiment. In some embodiments, the food product comprises meat or poultry substitute products selected from non-animal derived hamburgers, meatballs, sausages, meatloaf, kebab, and nuggets. Each possibility represents a separate embodiment. In some embodiments, the food product comprises fish products selected from burgers, nuggets, and sticks. Each possibility represents a separate embodiment. In some embodiments, the food product comprises fish substitute products selected from non-animal derived burgers, nuggets, and sticks. Each possibility represents a separate embodiment. In some embodiments, the food product comprises snack foods selected from chips and crackers. Each possibility represents a separate embodiment. In some embodiments, the food product comprises sauces selected from ketchup and mayonnaise. Each possibility represents a separate embodiment.

[0153] In some embodiments, the food product or food article is designated for non-human animal consumption (feed). Examples of animals are non-ruminants and ruminants. Ruminant animals include, for example, animals such as sheep, goats, cattle, e.g., beef cattle, cows, and young calves, deer, yank, camel, llama and kangaroo. Non-ruminant animals include mono-gastric animals, e.g., pigs or poultry such as turkeys, ducks and chicken; horses, young calves; fish; and crustaceans (including, but not limited to, shrimps andprawns). Animal feed for a monogastric animal typically comprises concentrates as well as vitamins, minerals, enzymes, direct fed microbial, amino acids and / or other feed ingredients (such as in a premix) whereas animal feed for ruminants generally comprises forage (including roughage and silage) and may further comprise concentrates as well as vitamins, minerals, enzymes, direct fed microbial, amino acid and / or other feed ingredients (such as in a premix). Typically, the animal feed is found in one of two forms: mash feed composed of all diet components mixed together or pelleted feed where the different diet components are compressed down into pellets with roughly the same size. In some embodiments, the food product or food article is an animal food product or article. In specific embodiments, the food product or food article is a pet food product or article designated for dogs, cats, and the like.

[0154] In some embodiments, methods of whitening a food product are provided, which methods comprising admixing the food product or a premix or intermediate thereof with the protein isolate compositions or the edible or emulsion compositions disclosed herein thereby whitening the food product. The term “whitening” as used herein refers to a change in color on the L*a*b* color scale to values that approach a reference white or a target white as detailed hereinabove. In some embodiments, the protein isolate compositions or the edible or emulsion compositions disclosed herein are characterized by white tones or colors and when admixed with the food product impart the white color to the food product thereby whitening it. Advantageously, imparting the white color to the food product is performed without oxidizing, reducing, or chemically reacting with the food product or ingredient thereof. Thus, in some embodiments, whitening the food product by admixing the food product or a premix or intermediate thereof with the protein isolate compositions or the edible or emulsion compositions disclosed herein does not comprise bleaching (i.e., oxidizing) the food product or ingredients thereof.

[0155] All scientific and technical terms used herein have meanings commonly used in the art unless otherwise specified. The definitions provided herein are to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0156] In the discussion unless otherwise stated, adjectives such as “substantially” and “about” modifying a condition or relationship characteristic of a feature or features of an embodiment of the invention, are understood to mean that the condition or characteristic is defined to within tolerances that are acceptable for operation of the embodiment for an application for which it is intended. Unless otherwise indicated, the word “or” in the specification and claims is considered to be the inclusive “or” rather than the exclusive or, and indicates at least one of, or any combination of items it conjoins.

[0157] It should be understood that the terms “a” and “an” as used above and elsewhere herein refer to “one or more” of the enumerated components. It will be clear to one of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Therefore, the terms “a”, “an”, and “at least one” are used interchangeably in this application.

[0158] For purposes of better understanding the present teachings and in no way limiting the scope of the teachings, unless otherwise indicated, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about”. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. As used herein the term “about” refers to ± 10 %.

[0159] In the description and claims of the present application, each of the verbs, “comprise”, “include”, and “have” and conjugates thereof, are used to indicate that the object or objects of the verb are not necessarily a complete listing of components, elements or parts of the subject or subjects of the verb. As used herein, the terms “consisting essentially of’ or “consists essentially” allow for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited.

[0160] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily skilled in the art upon examination of the followingexamples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.EXAMPLES

[0161] Generally, the nomenclature used herein, and the laboratory procedures utilized in the present invention include chemical, molecular, biochemical, and cell biology techniques. Such techniques are thoroughly explained in the literature. See, for example, “Molecular Cloning: A laboratory Manual” Sambrook et al., (1989); “Current Protocols in Molecular Biology” Volumes I-III Ausubel, R. M., ed. (1994); “Cell Biology: A Laboratory Handbook”, Volumes I-III Cellis, J. E., ed. (1994); The Organic Chemistry of Biological Pathways by John McMurry and Tadhg Begley (Roberts and Company, 2005); Organic Chemistry of Enzyme-Catalyzed Reactions by Richard Silverman (Academic Press, 2002); Organic Chemistry (6thEdition) by Leroy “Skip” G Wade; Organic Chemistry by T. W. Graham Solomons and, Craig Fryhle.

[0162] The present invention relates to a unique plant protein composition isolated from various legume sources such as pea (e.g., green pea, yellow pea), soy, lentils, and chickpea and its functional use in food applications, particularly as a food whitening agent.

[0163] Specific protein markers of the plant protein composition were analyzed by LC- MS / MS and SDS-PAGE assay. The plant protein composition obtained by the described method demonstrated superior techno-functional properties compared with those of commonly used plant-derived protein isolates thereby rendering it advantageous for use in several food applications.General MethodsExtraction and preparation of pea protein powder

[0164] Pea flour was first dispersed in deionized water for 3 h at a pH 7 to extract proteins into the aqueous phase. The solution was then centrifuged to remove insoluble components. The supernatant was then adjusted to the isoelectric point of pea proteins, pH 4.5, and aggregated globulins were separated from the albumin fraction by centrifugation. The supernatant (SI) was brought to 40 % saturation with ammonium sulfate. The solution wasallowed to stand for at least an hour and the first precipitate (Pl) was centrifuged at 5,000g. The supernatant (SII) was brought to 60% saturation with respect to ammonium sulfate and allowed to stand for two hours. The resulting precipitate (P2) was centrifuged at 16,000g for 15 min. Precipitate Pl was dissolved in 50 mM sodium phosphate buffer pH 6.8, and precipitate P2 was separately dissolved in 0.05M TRIS HC1 buffer pH 8.

[0165] Pl dissolved in 50 mM sodium phosphate buffer was applied to a diethylaminoethyl (DEAE)-Sepharose fast flow (Sigma- Aldrich) column (2.6 x 35 cm) and eluted with a linear salt gradient, increasing from 50 to 400 mM NaCl in 50 mM sodium phosphate buffer, pH 6.8, in a total volume of 800 mL. Fractions of 6 mL were eluted from the column at a flow rate of 20-25 mL / h. Individual peaks were analyzed by SDS-PAGE and the fractions containing protein with Mw at around 97 kDa were pooled.

[0166] P2 extract was chromatographed on a DEAE-Sepharose fast flow in 0.05M Tris HC1 with a linear gradient of 0 to 0.30 M NaCl and a flow rate of 10-15 ml / h. Individual peaks were analyzed by SDS-PAGE and peaks containing proteins with Mw at around 24- 26 kDa and at around 10 kDa were separately pooled.

[0167] The separated protein fractions were then mixed to form a pea protein extract. Aggregation of the pea proteins was performed by thermal treatment of the pea protein extract at a temperature range of 85-95°C. These aggregates were separated from the liquid phase by centrifugation (or vacuum filtration) followed by lyophilization to obtain an isolate in powder form.

[0168] Alternatively, proteins were extracted from pea flour, as disclosed hereinbelow. Pea flour was mixed with deionized water at a neutral pH to obtain an aqueous extract of plant proteins. A hydrocolloid (e.g., hydrocolloid-forming polysaccharide such as pectin, carrageenan, gelatin, gellan, hyaluronic acid, chitosan, a cellulose, e.g., carboxymethyl cellulose (CMC), a gum, e.g., locust bean gum, xanthan gum, Konjac gum, gellan gum, gum Arabic, and the like, alginate, agar or starch) was mixed with the aqueous extract to substantially encapsulate globulin proteins and water-insoluble plant matter. Subsequently, the aggregated encapsulate was separated from the aqueous extract. The resulting supernatant was mainly composed of plant albumins, which were precipitated to obtain the isolate by a method selected from salting out (e.g., by the addition of saturated ammoniumsulfate solution), ethanol-based coagulation, or by thermal treatment. Finally, the resulting precipitate was centrifuged and dried to obtain a white powder isolate.Evaluation of protein composition by SDS polyacrylamide gel electrophoresis (SDS- PAGE)

[0169] Polypeptides of pea soy and / or protein isolates were compared with the polypeptides of reference pea and soy isolates by SDS-PAGE assay. Pea reference extracts and isolates and protein extracts and isolates of the invention were prepared from the same source of commercial pea pulses (AGT, Canada). Reference pea extract was obtained by green pea flour extraction for 3 h at pH 9, followed by centrifugation to remove pea flour solids. GPP extract according to embodiments of the present invention was obtained from the green pea flour by a method comprising hydrocolloid-mediated globulin separation. Powder (isolate) forms were obtained from reference and invented extracts by a concentration or precipitation step as described above, including e.g., thermal treatment, centrifugation and lyophilization. Soy protein isolate was extracted from commercial soy flour (Dagesh, Israel). Commercial soy protein isolate (Solpro 922, China) was used as a reference soy isolate.

[0170] Precast Novex “WedgeWell” Tris-Glycine gels, 8-16% and Min Gel Tank (Invitrogen) were used for SDS-PAGE assay according to manufacturer recommendations. SDS-PAGE were prepared in reducing conditions. Molecular weight protein marker from Thermo Scientific (PageRuler Prestained Protein Ladder, Mw 10 to 180 kDa) was used. The staining of the gels was performed with Imperial Protein stain solution (Thermo Scientific). Band analysis and quantification was performed using BioRad Image Lab software. All detected bands in each lane were selected, and relative quantities (% band) were calculated according to the intensity and size of the bands.Relative protein concentrations in the extract using mass spectrometry-based quantitative proteomics (iBAQ) assay

[0171] Proteins were identified by using Discoverer 2.4 program with the Sequest (Thermo) algorithm. The samples of protein extracts or SDS-PAGE gel -excised bands were digested by trypsin, analyzed by LC-MS / MS on Q-Exactive plus (Thermo) and identified by Discoverer software against the Pisum sativum proteome from the Uniprot database, and adecoy database (in order to determine the false discovery rate) with a minimum of 2 peptides. High confidence peptides have passed the 1 % FDR threshold.

[0172] An intensity-based absolute quantification (iBAQ) method was used to estimate a protein amount. In this method, the intensities of all identified peptides of a protein are summed up, divided by the number of measurable tryptic peptides. iBAQ value reflects the absolute protein abundance in a sample. Proteomic assay of the extract obtained by the invented method detected 244 proteins. The relative amount of 16 proteins which are presented in substantial amounts are presented.Comparative color value of protein isolates vs. commercial protein isolates and titanium dioxide (gold standard)

[0173] Color measurements were performed with a ColorFlex EZ spectrophotometer (HunterLab). The CIELAB color system was used for color characterization. In this color space, the color points are characterized by three color coordinates. L * is the lightness coordinate ranging from no reflection for black (L * =0) to perfect diffuse reflection for white (L * =100). a * is the redness coordinate ranging from negative values for green to positive values for red. b * is the yellowness coordinate ranging from negative values for blue and positive values for yellow. The total color change is given by the color difference (* AEab), in terms of the spatial distance between two color points interpreted in the color space: (Hunter, 1987). If AE*ab <2.7, then the color difference between two samples can hardly be visually distinguished.Particle size measurement

[0174] The particle size distribution of powdered protein isolates was determined using a particle size analyzer (Mastersizer 2000, Malvern Inc., Worcestershire, United Kingdom) with a wet feed attachment. Samples were dispersed in deionized water (1% w / v). The particle size distribution was determined directly after ultrasound treatment or without treatment. The average particle sizes (D[3,2] - Surface Area Moment Mean - Sauter Mean Diameter and D[4,3] - Volume or Mass Moment Mean - De Brouckere Mean Diameter) and width of the distribution or Relative Span Factor (RSF) were analyzed.EXAMPLE 1 - GREEN PEA PROTEIN (GPP) COMPOSITIONS

[0175] A high content protein fraction from a pea flour, termed “Gavan green pea protein (GPP) extract” (or alternatively; “Gavan mixed extract”) was extracted and isolated. First, the Gavan GPP extract was analyzed by SDS-PAGE.

[0176] Three extracts obtained from pea flour were used for the analysis: an extraction of total proteins performed at pH 9.0 for three hours, followed by centrifugation to remove pea flour solids (termed “full extract”), an albumin extraction containing the albumin fraction of the full extract, extracted by traditional methods, e.g., globulin precipitation at acidic pH (termed “albumin fraction”), and the Gavan GPP extract (Fig. 1A). As shown in Fig. 1A, the detected proteins, with a relative abundance higher than 1%, can be divided into four main groups according to their molecular weight (Mw): group I: 5 kDa - 17 kDa, group II: 85 kDa -100 kDa, group III: 23 kDa - 26.9 kDa, and group IV: 27 kDa - 35 kDa. SDS-PAGE analysis revealed increased relative content levels of the larger proteins with Mw in the range of 85 kDa and 100 kDa (group II) in the Gavan GPP extract, compared to the albumin extract. Moreover, there was substantial increased content of the intermediate proteins, exhibiting Mw in the range between 23 kDa and 26.9 KDa (group III), in the Gavan GPP extract vs. the albumin extract.

[0177] To further characterize the protein content of the Gavan GPP extract, it was analyzed by liquid chromatography with tandem mass spectrometry (LC-MS / MS). The results are summarized in Table 1 below, in which the values reflect absolute protein abundance in the sample. Proteomic assay of both extracts detected 244 proteins. Relative amounts of 16 proteins which are present in substantial amounts (>1%) are presented in the table.Table 1 - Mass spectrometry-based relative protein quantification (iBAQ) of two extracts: albumin fraction of pea protein extract at pH 9 vs. Gavan GPP extract

[0178] As shown in Table 1, comparison between the Gavan GPP extract and the albumin extract, by the LC-MS / MS analysis, revealed some substantial differences, even though the total protein fraction of the 16 most abundant proteins remained similar (-89% and 92% in the Gavan GPP extract and the albumin extract, respectively). In particular, the relative content of the most abundant protein among the proteins in the albumin extract having a Mw of 10.6 kDa, namely seed albumin protein of late embryogenesis 1 abundant group 1, wasreduced in the Gavan GPP extract compared to the albumin extract (14.7% in the Gavan GPP extract vs. 27% in the albumin extract). In fact, the most abundant protein in the Gavan GPP extract was found to be albumin-2 having a Mw of 26.2 kDa (17.8% in the Gavan GPP extract vs. 12.8% in the albumin extract).

[0179] Table 2 summarizes the relative content of each protein, sorted by its Mw size, from low to high. As demonstrated in Table 2, Gavan GPP extract comprised reduced small proteins content with Mw in the range of 5 kDa - 17 kDa, though their fraction was still above 50% of the total protein content (-55.4%). LC-MS / MS results supported the SDS- PAGE analysis, demonstrating increased proteins content exhibiting Mw within the range of 23 - 26.9 kDa, and 85 -100 kDa (19.8% and 3.5% for the Gavan GPP extract vs. 15.9% and 0.9% in the albumin extract, respectively). Among the larger proteins group, the relative contents of both seed linoleate 9s-lypoxygenase-2 and seed linoleate 9s-lypoxygenase-3 were found to be higher in the Gavan GPP extract.

[0180] Overall, the relative contents of all albumin proteins (e.g., seed albumin late embryogenesis 1, albumin 2, albumin 1A, albumin ID, albumin IE, Albumin IF, and seed albumin 2-fragment) were similar in the Gavan GPP extract and the albumin fraction (52.8% in the Gavan GPP extract vs. 57.5 in the albumin fraction control).Table 2 - LC-MS / MS analysis of Gavan GPP extract of a green pea* Values marked in bold represent increased relative abundance in the Gavan GPP extract.

[0181] SDS-PAGE was then performed on the Gavan GPP powder (isolate) compared to commercial pea protein isolates (termed “commercial 1 and commercial 2”) (Fig. IB). Quantification analyses based on the SDS-PAGE of Fig. IB revealed that whereas the ratio of a reference protein fraction (having a molecular weight between 40 kDa and 70 kDa) to the protein fraction having a molecular weight between 85 kDa and 105 kDa of the isolate of the present invention was in the range of 0.02-0.9, the commercial isolates were characterized by ratios of 3.2 and 5 (commercial 1 and commercial 2, respectively).

[0182] Nutritional analysis of the Gavan GPP powder (isolate), compared to commercial protein isolates from pea, composed of 80% proteins, is demonstrated in Tables 3 and 4. As shown in these tables, substantial nutritional differences were revealed and overall, there was superiority to the Gavan GPP powder in comparison to commercial products, specifically in terms of protein composition and quality.Table 3 - Amino acid content in GPP compared to commercial pea isolates* Values marked in bold represent a positive delta in the specified amino acid content of a GPP compared to a commercial product.

[0183] As shown in Table 3, Gavan GPP powder was found to have a higher content of total amino acids compared to the commercial products (delta = 10-29%). Specifically, there were increased levels of essential amino acids (EAAs) in the Gavan GPP isolate. The contents of the EAAs: isoleucine, lysine, methionine, threonine, valine, and tryptophan, were elevated in the Gavan GPP isolate by 21-47% compared to the commercial products. The levels of the branched amino acids (BCAAs): leucine, isoleucine, and valine, were elevated by 10-38%. Among the EAAs, there were exceptionally high contents of the EAAs: lysine (delta = 46-57%), methionine (delta = 48-63%), threonine (delta = 58-92%) and valine (delta = 43-63%).

[0184] Excessive content of glutamic acid in food products, in addition to its known off- flavor in plant proteins, can be harmful. The Gavan GPP powder was found to have a lower content of glutamic acid by 21-29%, compared to the commercial products, indicating an additional nutritional advantage.

[0185] Table 4 summarizes the analytic nutritional analysis of the Gavan GPP powder, compared to a commercial product.Table 4 - Analytic nutritional analysis of Gavan GPP isolate as compared to control

[0186] As shown in Table 4, the Gavan GPP isolate and the commercial product demonstrated similar total protein content (82.7% and 80%, respectively). However, a reduced fat content was found in the Gavan GPP isolate compared to a commercial product (1.4% vs. 9%, respectively). Although there was increased carbohydrates content in the Gavan GPP isolate, the Kcal / lOOg value was found to be reduced in the Gavan GPP isolatecompared to the commercial product (371 Kcal / lOOg in the Gavan GPP isolate vs. 400 Kcal / lOOg in the commercial product).

[0187] One of the most dramatic differentiating properties of the Gavan GPP isolate was found to be its bright white color, as seen in both powder form (following thermal treatment and lyophilization, Fig. 2A) and wet form (power isolate as described above that was dispersed in water, Fig. 2B). This desired characteristic results from the unique plant protein composition of the Gavan GPP powder and particle size.

[0188] Measurement of color coordinates was performed using Hunter Lab Easy Match QC software at D65 illumination which corresponds to an average daylight. “Gold standard white” values are at approximately: L*: 95-100, a*: -0.1 to 0.1, and b*: -0.1 to 0.1. The results of the color coordinates of Gavan GPP powder compared to commercial pea protein isolates are presented in Table 5 below and in Fig. 3, where “1” indicates Green pea isolate S85F plus D, “2” indicates Green pea isolate S85F, “3” indicates pea isolate 85 protein, and “4” indicates the Gavan GPP isolate powder.Table 5 - Color coordinates for Gavan GPP powder compared to commercially available pea protein isolates

[0189] As shown in the table and figure, the Gavan GPP powder showed values of L*, a* and b* which are closer to a reference white color. The L* value was 93.96 and the a* value was -0.11, which are almost in the range of a reference white. The b* value of Gavan's GPP powder was 5.96, which is at least 3.5-fold closer to a reference white compared to commercial pea protein products.

[0190] The whiteness trait of the Gavan GPP powder was then compared to titanium dioxide (TiO2) particles with sizes of -250 nmused in the food industry as a whitening agent.

[0191] As demonstrated in Fig. 4 and Table 6 below, it was unexpectedly found that the Gavan GPP isolate showed superior whitening potential, even when its particle sizes were 2-3 orders of magnitude larger than the particles sizes of TiO2.Table 6 - Color coordinates and delta E of Gavan powders compared to reference commercial products

[0192] Comparison between the TiO2 powder, Gavan GPP powder, and two other types of commercially available plant protein powders (Fig. 4), demonstrated the feasibility of replacing TiO2 by protein isolates of the invention, such as the Gavan GPP powder. Examination of the delta E (dE) value, which is generally used to estimate the proximity between two subjected colors, was found to be 3.284, between the Gavan GPP powder and the TiO2 powder, indicating that the difference between the colors of the Gavan GPP powder and the TiO2 is almost indistinguishable by a human eye (dE<2.7 is considered indistinguishable). Commercial protein isolate powders, even at 90% purity, were completely off the indistinguishable range, having almost 5-fold increased dE value, compared to the Gavan GPP powder. All commercial samples had significantly lower values of L* lightness coordinate and markedly higher values of yellowness and redness coordinates.

[0193] The potential of the Gavan GPP powder as a whitening agent was further examined in water suspensions, composed of 1% and 1.5% (w / v) Gavan GPP. As shown in Table 7, results of the suspended Gavan GPP powder demonstrated high applicative compatibility, as the dE value between the 1% Gavan GPP suspension and the TiO2 suspension was found to be 3.06, indicating a decrease of about 7% in dE, as compared to the powder analysis.Table 7 - CIELAB color coordinates and dE of Gavan GPP (i.e., plant-protein isolate of the invention) suspensions compared to TiOi suspensions

[0194] Examination of particle size results adds another dimension to the grade of whiteness of Gavan GPP powder, as the whiteness trait is dependent upon particle size. The results are shown in Figs. 5A-5B and Table 8 below.Table 8 - Particle size distribution of the Gavan GPP extract powder

[0195] As shown in Figs. 5A-5B and Table 8, the Gavan GPP powder has a particle size of Dv(50) = 47.6 pm, in its powder form (Fig. 5A). After 5 seconds of sonication, the particle size was reduced by approximately 77% to Dv(50) = 11 pm (Fig. 5B), demonstrating feasibility in integrating the product in different food applications, for different uses (e.g., colorant, texture modifier, flavoring agent, etc.).EXAMPLE 2 - SOY PROTEIN (SP) COMPOSITIONS

[0196] The features of a protein isolate obtained from soy seeds by the same extraction and processing technique as the Gavan GPP powder (termed herein: “Gavan soy protein (SP)”) was examined. To this end, samples of Gavan SP isolates in powder form were mixedwith sample buffer, and subjected to SDS-PAGE analysis, essentially as described in Example 1.

[0197] Unexpectedly, the Gavan soy protein isolate showed more dramatic changes in its protein composition compared to a commercial soy isolate, as examined by SDS-PAGE analysis (Fig. 6). Even though soy is known to naturally contain less albumins compared to peas, the Gavan SP isolate was found to obtain similar features to the Gavan GPP isolate. As shown in Fig. 6, similarly to the Gavan GPP isolate, there was increased relative content of proteins at Mw of 85-100 kDa and 23-27 kDa. Additional differences were observed at Mw of 27-35 kDa and at Mw of 35-70 kDa, which were more visually dominant compared to the Gavan GPP isolate. In contrast to the Gavan GPP isolate, a band at Mw size of ~45 kDa appeared in the Gavan SP isolate and was absent in the commercial SP product.

[0198] Quantification of the SDS-PAGE showed that the ratio of a reference protein fraction (having a molecular weight between 40 kDa and 70 kDa) to the protein fraction having a molecular weight between 85 kDa and 105 kDa in a soy isolate according to embodiments of the present invention was in the range of 0.9- 1.7. In contrast, the ratio of a reference protein fraction (having a molecular weight between 40 kDa and 70 kDa) to the protein fraction having a molecular weight between 85 kDa and 105 kDa in a commercial soy isolate (Solpro 922) was 7.2.

[0199] Color coordinates and reflectance / transmittance (R / T) for Gavan SP and GPP compositions (two samples each provided during isolation) is shown in Table 9 below. Particle size distribution of the resulting powders before and after sonication are shown in Figs. 7A-7B and Table 10 below.Table 9 - Color coordinates and R / T of Gavan SP and GPP

[0200] Examination of CIELAB color coordinates of the Gavan SP isolate, demonstrated highly whitening feature, similarly to Gavan's GPP. Measured color coordinates for thetested Gavan SP isolates were L* = 93.43, a* = 1.29, and b* = 6.46; and L* = 94.89, a* = 0.14, and b* = 5.79.Table 10 - Particle size distribution of Gavan SP powder

[0201] The Gavan SP powder showed similar particle distribution to the Gavan GPP powder, as shown in Figs. 7A-7B. The Gavan SP powder was found to have a particle size characterized by Dv(50) = 42 pm, in its powder form (Fig. 7A). After 5 seconds of sonication, the particle size was reduced by about 68% to Dv(50) = 13.4 pm (Fig. 7B).EXAMPLE 3 - RED LENTILS PROTEIN (RLP) COMPOSITIONS

[0202] A protein extract was obtained from red lentil seeds (termed herein: “Gavan red lentils protein (RLP)”) and samples of Gavan RLP isolates in powder form were mixed with sample buffer, and subjected to SDS-PAGE analysis, essentially as described in Example 1.

[0203] Similar to the Gavan GPP isolate, and to the Gavan SP isolate, there was increased appearance of proteins at Mw within the range of 85-100 kDa indicating a repeated presence of these large proteins in all examined Gavan isolates. Additional differences were observed at Mw of 20-35 kDa and at Mw of 50-70 kDa. In the absence of a commercial red lentil isolate, Gavan's red lentil isolate was compared to a full extract of red lentil proteins obtained by an extraction of the total proteins at pH 9.0 for three hours, followed by centrifugation to remove lentil flour solids. As shown in Fig. 8, a clear reduction in the amount of proteins having a molecular weight between 40 kDa and 70 kDa was noticed as compared to the full extract. Quantification of the SDS-PAGE revealed that the ratio of a reference protein fraction (having a molecular weight between 40 kDa and 70 kDa) to the protein fractionhaving a molecular weight between 85 kDa and 105 kDa in the RLP isolate of the present invention was 1.34.

[0204] Next, the amino acids distribution in the Gavan RLP powder was analyzed (Table 11). As shown in the table, Gavan RLP was found to possess superior nutritional advantages. In particular, Gavan RLP powder was found to have a higher content of total amino acids compared to the GPP commercial products (delta = 19-40%). Gavan RLP powder showed an increase in the EAAs content (33-61%), and reduced levels of glutamic acid compared to the commercial pea protein products (delta = 5-14%). The Gavan RLP powder was also shown to contain exceptionally increased levels of BCAAs vs. the commercial products. Table 11 - Amino acid content in RLP isolate compared to commercial pea protein isolates* Values marked in bold represent a positive delta in the specified amino acid content of a RLP isolate of the invention compared to a commercial pea product.

[0205] Fig. 9 and Table 12 demonstrate the optical properties of the Gavan RLP powder. Table 12 - Color coordinates of Gavan RLP compositions

[0206] As shown in Fig. 9 and Table 12, the Gavan RLP powder exhibits a substantial whitening potential, similar to the Gavan GPP and the Gavan SP samples.

[0207] Figs. 10A-10B and Table 13 show the particle size distribution.Table 13 - particle size distribution of Gavan RLP powder

[0208] As can be seen in Figs. 10A-10B and Table 13, the Gavan RLP powder was found to have a particle size characterized by Dv(50) = 43.6 pm, in its powder (isolate) form (Fig. 10A). After 5 seconds of sonication, the particle size was reduced by about 56% to Dv(50) = 19.4 pm (Fig. 10B).

[0209] Table 14 summarizes the CIELAB color coordinates and dE of the Gavan GPP powder, the Gavan SP powder, and the Gavan RLP powder, compared to the gold standard TiO2, and in reference to commercial pea and soy isolates.Table 14 - Color coordinates and delta E of Gavan powders compared to the gold standard Tith, and the reference commercial products

[0210] As shown in Table 14, all examined Gavan GPP powder, Gavan SP powder andGavan RLP powder, exhibited superior dE values (-6-10 fold) compared to the commercial products. In fact, the dE of the Gavan SP powder was 1.73, indicating that this powder in visually identical to TiO2. Thus, Gavan's isolates from soy, green pea, and red lentils all exhibited remarkable shade characteristics that are advantageous as food whitening additives.

[0211] Table 15, herein below, summarizes the particle size before (time 0) and after 5 seconds (time 5) of sonication of Gavan's isolates.Table 15 - Specific surface area and Dv of Gavan GPP, SP and RLP powders

[0212] As shown in Table 15, the particle size distribution analysis of Gavan's isolates showed narrow and uniform size distributions indicating that Gavan's powders are suitable for incorporation into a variety of food products.

[0213] In summary, the compositions according to embodiments of the present invention were characterized by a ratio of a reference protein fraction (having a molecular weight between 40 kDa and 70 kDa) to the protein fraction having a molecular weight between 85 kDa and 105 kDa of less than 2.25 while the ratios measured in hitherto known compositions (such as commercial protein isolates) were significantly higher. The compositions according to embodiments of the present invention were further characterized by superior white color that most resembles the gold standard white of TiO2. Finally, the compositions in powder form exhibited narrow and uniform particles size distributions that are particularly advantageous for use in the food industry.

[0214] While the present invention has been particularly described, persons skilled in the art will appreciate that many variations and modifications can be made. Therefore, the invention is not to be construed as restricted to the particularly described embodiments, and the scope and concept of the invention will be more readily understood by reference to the claims, which follow.

Claims

CLAIMSWhat is claimed is:

1. A food whitening additive comprising a plant protein isolate derived from a legume extract, wherein the plant protein isolate has a protein content of at least 80% and is characterized by:(i) a first protein fraction having a molecular weight between 5 kDa and 17 kDa,(ii) a second protein fraction having a molecular weight between 85 kDa and 105 kDa in a weight portion relative to the total plant proteins of the plant protein isolate of at least 1%, and(iii) a weight ratio of a reference protein fraction having a molecular weight between 40 kDa and 70 kDa, if present, to the second protein fraction, of 2.25 or less, and wherein the plant protein isolate is further characterized by a total color difference (AE) in CIELAB coordinates of less than 4, relative to titanium dioxide powder.

2. The food whitening additive of claim 1, wherein the plant protein isolate is characterized by L* CIELAB value in a range between about 90 and about 95; a* CIELAB value in a range between about (-0.1) and about (0.4); and b* CIELAB value in a range between about 4.0 and about 10.0.

3. The food whitening additive of claim 1, wherein the plant protein isolate is characterized by L* CIELAB value in a range between about 80 and about 95; a* CIELAB value in a range between about (-2) and about 1.5; and b* CIELAB value in a range between about 3 and about 10.0.

4. The food whitening additive of any one of claims 1 to 3, wherein the plant protein isolate is in the form of a powder.

5. The food whitening additive of claim 4, wherein the plant protein isolate is characterized by Dv(50) between about 10 pm and about 20 pm.

6. The food whitening additive of claim 4, wherein the plant protein isolate is characterized by Dv(50) between about 35 pm and about 55 pm.

7. The food whitening additive of claim 4, wherein the plant protein isolate is characterized by at least one of: uniformity of between about 0.6 and about 1.2; span of between about 2 and about 3.7; D[3,2] of between about 4 pm and about 9 pm; and D[4,3] of between about 15 pm and about 25 pm.

8. The food whitening additive of claim 4, wherein the plant protein isolate is characterized by at least one of: uniformity of between about 0.6 and about 1.2; span of between about 2 and about 3.7; D[3,2] of between about 15 pm and about 20 pm; and D[4,3] of between about 50 pm and about 65 pm.

9. The food whitening additive of claim 4, wherein the plant protein isolate is characterized by a specific surface area of at least 300 m2 / kg.

10. The food whitening additive of claim 9, wherein the plant protein isolate is characterized by a specific surface area of at least 700 m2 / kg.

11. The food whitening additive of any one of claims 1 to 10, wherein the second protein fraction having a molecular weight between 85 kDa and 105 kDa comprises a lipoxygenase.

12. The food whitening additive of claim 11, wherein the lipoxygenase is enzymatically inactive.

13. The food whitening additive of any one of claims 1 to 12, wherein the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 3%.

14. The food whitening additive of claim 13, wherein the second protein fraction having a molecular weight between 85 kDa and 105 kDa is in a weight portion relative to the total plant proteins of the plant protein isolate of at least 10%.

15. The food whitening additive of any one of claims 1 to 14, wherein the plant protein isolate has an albumin content of at least 20% w / w.

16. The food whitening additive of claim 15, wherein the plant protein isolate has an albumin content of at least 30% w / w.

17. The food whitening additive of claim 16, wherein the plant protein isolate has an albumin content of at least 40% w / w.

18. The food whitening additive of any one of claims 1 to 17, wherein the plant protein isolate is substantially devoid of globulins.

19. The food whitening additive of any one of claims 1 to 18, wherein the plant protein isolate is substantially devoid of the reference protein fraction.

20. The food whitening additive of any one of claims 1 to 19 comprising trace amounts of a hydrocolloid.

21. The food whitening additive of any one of claims 1 to 20, wherein the plant protein isolate is derived from seeds of a legume plant.

22. The food whitening additive of any one of claims 1 to 21, wherein the legume is selected from the group consisting of pea, soy, chickpea, lentil, bean, peanut, and any combination thereof.

23. The food whitening additive of claim 22, wherein the legume is green pea.

24. The food whitening additive of claim 22, wherein the legume is yellow pea.

25. The food whitening additive of claim 22, wherein the legume is soy.

26. An edible composition comprising the food whitening additive of any one of claims 1 to 25.

27. The edible composition of claim 26, comprising at least 0.1% w / w of the food whitening additive.

28. The edible composition of claim 26 or 27, in the form of a water-in-oil emulsion or an oil-in-water emulsion.

29. The edible composition of claim 28, in the form of an oil-in-water emulsion comprising 0.03-3% w / w of the food whitening additive; 20%-98%w / w of an oily phase; and 2-80% w / w of an aqueous phase.

30. A food product comprising between 0.1% and 30% w / w of the food whitening additive of any one of claims 1 to 25.

31. The food product of claim 30, wherein the food product comprises between 0.1% and 5% w / w of the food whitening additive.

32. A method of whitening a food product, comprising combining the product or a premix or intermediate thereof with the food whitening additive of any one of claims 1 to 25 or the edible composition of any one of claims 26 to 29.

33. The method of claim 32, wherein the food product is suitable for human consumption or for non-human animal consumption.

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