A method for preparation of alternative dairy products

Abstract

A food product comprising a modified plant-based protein, fat and a polysaccharide is provided, wherein the polysaccharide is selected from a polysaccharide containing hydrocolloids, modified starch, native starch, or any combination thereof. Methods for manufacturing the food product are also provided.

Description

A METHOD FOR PREPARATION OF ALTERNATIVE DAIRY PRODUCTSCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority under 35 USC §119(e) of U.S. Provisional Patent Application No. 63 / 553,701, filed February 15, 2024, entitled “A METHOD FOR PREPARATION OF ALTERNATIVE DAIRY PRODUCTS”, the contents of which are incorporated herein by reference in their entirety.FIELD OF THE INVENTION

[0002] The invention relates generally to the field of plant-based cheese and other dairy products.BACKGROUND

[0003] Dairy proteins, particularly casein and its derivatives, have a unique composition and molecular structure with characteristics that determine their functional properties of various dairy products, such as cheese and yogurt. Mostly, dairy proteins that are used in the industry have specific properties, which contribute to the textural properties of dairy food. There are only a few economically viable examples of plant proteins that exhibit textural characteristics of dairy proteins used for such purposes. Using fiber-like vegetable proteins as alternatives to dairy protein-based food products is highly challenging.

[0004] Common plant proteins used in plant-based foods today include soy, peas, potatoes, mung beans, and rice, all distinguished with their globular structures.

[0005] Biopolymers, mainly proteins and polysaccharides can produce gels that can be used to create a desired texture for food formulations in general, and for cheese, and yogurts analogues, and they can bind to each other, thus creating a porous three-dimensional network of biopolymer chains that can trap water, oil / fat and other substances in it. Today, starches are mainly used to form the three-dimensional network of biopolymer chains, but several concerns have emerged regarding their use. Some of the concerns relate to high carbohydrate content and no nutritional value of these products due to the lack of protein in commercially available plant-based dairy products, particularly, cheeses and yogurts.SUMMARY

[0006] The following embodiments and aspects thereof are described and illustrated in conjunction with systems, tools and methods which are meant to be exemplary and illustrative, not limiting in scope.

[0007] In one aspect, there is provided a food product comprising (i) a modified protein, (ii) a polysaccharide and (iii) an oil; wherein: the (i) and the (ii) are non-crosslinked;

[0008] the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 5 and 40%; and optionally is further characterized by a molar ratio between glutamine and glutamic acid residues of below 1: 10;

[0009] the food product is a homogenous solid at a temperature below a melting onset point, and the melting onset point is at a temperature ranging between 30 and 40°C.

[0010] In another aspect, there is provided a method for manufacturing the food product of the invention, comprising:

[0011] contacting an aqueous composition comprising a protein with (a) an enzyme having a deamidase and a proteolytic activity; or (b) a first enzyme having a deamidase activity and with a second enzyme having a proteolytic activity, thereby obtaining a composition comprising the modified protein;

[0012] mixing the modified protein or the composition with the polysaccharide and oil, thereby obtaining the food product.

[0013] In another aspect, there is provided a method for manufacturing the food product of the invention, comprising:

[0014] contacting an aqueous composition comprising a protein with an enzyme having a proteolytic activity and optionally an additional enzyme having deamidase activity, thereby obtaining a composition comprising the modified protein;

[0015] mixing the modified protein or the composition with the polysaccharide and oil , thereby obtaining the food product.

[0016] In another aspect, there is provided a food product comprising (i) a modified protein, (ii) a polysaccharide and (iii) a fat; wherein: the (i) and the (ii) are non-crosslinked;the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 5 and 40% or between 0.5 and 30%; and optionally is further characterized by a molar ratio between glutamine and glutamic acid residues of below 1:10; the food product is a homogenous solid at a temperature below a melting onset point, and the melting onset point is at a temperature ranging between 30 and 40°C.

[0017] In one embodiment, in a non -hydrated state the food product comprises the modified protein at a concentration between 2 and 60%w / w, and the fat at a concentration between 5 and 50%w / w, relative to the dry weight of the food product.

[0018] In one embodiment, a weight ratio between the modified protein and the polysaccharide is between 1: 10 and 10:1, and wherein the homogenous solid is substantially devoid of phase separation.

[0019] In one embodiment, fat is or comprises an oil, saturated fatty acid, unsaturated fatty acid, fatty acid mono-, di- and / or tri-glyceride, fatty acid phosphate ester, and phospholipid, or any combination thereof.

[0020] In one embodiment, the polysaccharide is or comprises a modified starch, a native starch, or any combination thereof.

[0021] In one embodiment, the modified protein is derived from a plant protein; wherein the hydrolysis degree of the modified protein is between about 5 and about 20% or between about 1 and about 25%; and wherein the food product is a dairy food product substitute.

[0022] In one embodiment, the dairy food product substitute is a cheese substitute.

[0023] In one embodiment, the food product is characterized by at least one of: a hardness between 5 and 40N, an adhesiveness between 0.1 and 3Nmm, and a fracture between 5 and 30N, determined by Texture Profile Analyzer.

[0024] In one embodiment, derived is by exposing the plant protein to a first enzyme having a deamidase activity and optionally to a second enzyme having a proteolytic activity.

[0025] In one embodiment, the modified protein is characterized by at least one of: (i) alpha helix content of at least 50%; (ii) average MW below 120kDa; (iii) water solubility above 25%, as determined based on total nitrogen content; and wherein a weight ratio between the modified protein and the polysaccharide is between 1: 1 and 1:5.

[0026] In one embodiment, the food product in a hydrated state is characterized by a water content between 30 and 70% w / w, and wherein a weight content of the modified protein by dry weight of the food product is between about 10 and about 20%.

[0027] In one embodiment, the food product is characterized by meltability, wherein the meltability comprises a diameter increase between 170 and 350%, as determined by Schreiber test; wherein the polysaccharide is starch; optionally wherein a weight ratio between the modified protein and the polysaccharide is between 1: 1 and 1:3.

[0028] In another aspect, there is provided a method for manufacturing the food product of the invention, comprising:

[0029] contacting an aqueous composition comprising a protein with (a) an enzyme having a deamidase and a proteolytic activity; or (b) a first enzyme having a deamidase activity and with a second enzyme having a proteolytic activity, thereby obtaining the modified protein;

[0030] mixing the modified protein or the composition comprising thereof with the polysaccharide and fat, thereby obtaining the food product.

[0031] In one embodiment, the step (i) is performed at a temperature between 10 and 70°C; and wherein the step (ii) comprises mixing at a temperature between 40 and 110 °C for a period of time sufficient for forming the food product.

[0032] In one embodiment, step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:2 and 10: 1; optionally step (ii) further comprises adding one or more additional agents selected from flavoring agent, pigment, hydrocolloid, coloring agent, preservative, and salt.

[0033] In one embodiment, the aqueous composition is substantially devoid of the polysaccharide.

[0034] In one embodiment, the modified protein is characterized by a hydrolysis degree between 5 and 40% or between 0.5 and 30%; and by a molar ratio between glutamine and glutamic acid residues of below 1:10; and by at least one of: alpha helix content of at least 50%; average MW below 120kDa; water solubility above 25%, as determined based on total nitrogen content.

[0035] In one embodiment, a concentration of the protein and / or of the modified protein within the aqueous composition is between 3 and 30%w / w; and wherein the modified protein is characterized by a hydrolysis degree between 1 and 25%.

[0036] In one embodiment, at a concentration of the protein within the aqueous composition of about 20%w / w the aqueous composition is a non-flowable composition, and wherein the composition obtained after completion of step (i) is in a form of a flowable liquid.

[0037] In one embodiment, step (i) comprises adding to the aqueous composition 0.01%- 3%w / w of the first enzyme relative to the protein; optionally wherein the first enzyme comprises amidohydrolase, deamidase, protein glutaminase, or any combination thereof.

[0038] In one embodiment, the method further comprising performing a termination step thereby substantially arresting enzymatic activity of the a or of the b, wherein the termination step is performed prior to the step (ii).

[0039] In one embodiment, termination step comprises any of: heating the aqueous composition to a temperature of at least 90 °C; adding to the aqueous composition an inhibitor of the a or of the b; or exposing the aqueous composition to an acid to obtain a pH of below 4 or above 9.

[0040] In another aspect, there is provided a method for manufacturing the food product of the invention, comprising:

[0041] contacting an aqueous composition comprising a protein with an enzyme having a proteolytic activity and optionally an additional enzyme having deamidase activity, thereby obtaining an aqueous composition comprising the modified protein;

[0042] mixing the modified protein or the aqueous composition with the polysaccharide and fat, thereby obtaining the food product; and wherein the aqueous composition is substantially devoid of a polysaccharide.

[0043] In one embodiment, the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 3 and 40% or between 0.5 and 30%.

[0044] In one embodiment, the modified protein is further characterized by at least one of: alpha helix content of at least 50%; average MW below 120kDa; hydrolysis degree between 1 and 20%; and water solubility above 25%, as determined based on total nitrogen content.

[0045] In one embodiment, a concentration of the protein within the aqueous composition is between 3 and 30%w / w, and wherein the aqueous composition consists essentially of the protein.

[0046] In one embodiment, step (i) comprises adding to the aqueous composition between 0.01%-3%w / w of the enzyme relative to the protein.

[0047] In one embodiment, step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:2 and 10: 1.

[0048] In one embodiment, step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:3 and 3: 1.

[0049] In one embodiment, step (i) further comprises mixing at a temperature between 10 and 70°C.

[0050] In one embodiment, step (i) further comprises performing a termination step, wherein the termination step is performed prior to the step (ii).

[0051] In one embodiment, termination step comprises any of: providing the aqueous composition to a temperature of at least 90°C; adding an inhibitor of the enzyme to the aqueous composition; and exposing the aqueous composition to a pH below 4 or above 9.

[0052] In one embodiment, the step (i) further comprises isolating the modified protein from the aqueous composition.

[0053] In one embodiment, isolating is by drying the aqueous composition to obtain a powder comprising the modified protein.

[0054] In one embodiment, the step (ii) further comprises drying the food product, optionally wherein the drying is by lyophilization or spray drying.

[0055] In one embodiment, the polysaccharide is or comprises starch.

[0056] In one embodiment, the enzyme having a proteolytic activity comprises or is selected from any one of: pepsin, trypsin, chymotrypsin, papain, endoprotease, exoprotease, carboxypeptidase, aminopeptidase, protein deamidase or any combination thereof.

[0057] In one embodiment, the protein is or comprises any one of: a plant protein, a plant protein isolate, a concentrated wet plant protein and a precipitated wet plant protein.

[0058] In one embodiment, the modified protein has at least 15% greater water solubility compared to the protein, as determined based on total nitrogen content; and wherein the modified protein has a hydrolysis degree between about 2 and about 25%.

[0059] In one embodiment, the starch is or comprises a modified and / or non-modified starch, optionally wherein the starch comprises or is selected from any of: potato starch, cornstarch, rice starch and tapioca starch, or any combination thereof; and wherein a weight ratio between the modified protein and the polysaccharide is between 1: 1 and 1:5.

[0060] In another aspect, there is provided a modified protein derived from a plant protein, wherein the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 0.5 and 30% and by a molar ratio between glutamine and glutamic acid residues below 1:10; wherein the modified protein has a water solubility above 25%w / w, as determined based on total nitrogen content and is characterized by at least one of: (i) alpha helix content of at least 50%; and (ii) average MW below 120kDa.

[0061] In one embodiment, derived is by exposing the plant protein to a first enzyme having a deamidase activity and optionally to a second enzyme having a proteolytic activity.

[0062] In one embodiment, the modified protein is characterized by a hydrolysis degree between about 1.5 and about 25%; molar ratio between glutamine and glutamic acid residues between 1:10 and 1: 1000; alpha helix content of at least 50%; and by average MW below 80kDa.

[0063] In another aspect, there is provided aA food product comprising the modified protein of the invention, (ii) a polysaccharide and (iii) a fat; wherein: the (i) and the (ii) are non-crosslinked;

[0064] the food product is a homogenous solid at a temperature below a melting onset point, and the melting onset point is at a temperature ranging between 30 and 40°C.

[0065] In one embodiment, in a non-hydrated state the food product comprises the modified protein at a concentration between 2 and 60%w / w, and the fat at a concentration between 5 and 50%w / w; wherein a weight ratio between the modified protein and the polysaccharide is between 1: 1 and 1: 10, and wherein the homogenous solid is substantially devoid of phase separation.

[0066] In one embodiment, the food product is a cheese substitute and is characterized by at least one of: a hardness between 5 and 40N, a adhesiveness between 0.1 and 3Nmm, and a fracture between 5 and 30N, determined by Texture Profile Analyzer; and wherein the cheese substitute has a meltability comprising a diameter increase between 170 and 350%, as determined by Schreiber test.

[0067] In one embodiment, the cheese substitute is selected from semi-hard and hard cheese substitute.

[0068] In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to the figures and by study of the following detailed description.

[0069] 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.

[0070] 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 DRAWINGS

[0071] Fig. 1A-1B. is a bar graph presenting integration data of 1A C=O amide (at 1629 cm1) and IB carbonyl ester (at 1742 cm1) of PPI before and after treatment with protein glutaminase (PG) for 1 h.

[0072] Fig. 2 is a graph presenting transformation degree of glutamine to glutamic acid of a 10% wt pea protein isolate (PPI) in water after treatment with PG over time, results are expressed as means of triplicates with standard deviation.

[0073] Fig. 3A-3B. are graphs presenting the degree of hydrolysis of 10% wt of plant protein after treatment with papain over time. 3A. PPI mixture, results are expressed as means of triplicates with standard deviation and 3B. SPI mixture, results are expressed as means of duplicates with standard deviation.

[0074] Fig. 4A-4B. presents endoprotease activity of PG - deamidaze and endoprotease. 4A SDS-PAGE of PPI before (lane 1) and after PG treatment (lane 3 and 4). 4B is SEC MALS of SPI before (upper) and after PG treatment (lower).

[0075] Fig. 5A-5B. presents an image of a 20% wt in water of soy protein isolate (SPI) before and after treatment with PG.

[0076] Fig. 6. is an image presenting visual assessment of sandy texture in native PPI and SPI.

[0077] Figs. 7A-7D. are images of Modified Schreiber test for PPI cheese melting. Fig. 7A (from left to right): dairy cheese, marketed vegan cheese, native PPI. Fig. 7B (from left to right): PPI treated with PG for 0.5, 1.5 and 3h. Fig. 7C (from left to right): PPI treated with papain for 0.5, 1.5 and 3h. Fig. 7D (from left to right): PPI treated with Endo-Stern Enzyme Bp, Protease Neutral, Flavourzyme and Alcalase.

[0078] Figs. 8A-8D. are images of Modified Schreiber test for SPI cheese melting. Fig. 8A (from left to right): dairy cheese, marketed vegan cheese, native SPI. Fig. 8B (from left to right): SPI treated with PG for 0.5, 1.5 and 3h. Fig. 8C (from left to right): SPI treated with papain for 0.5, 1.5 and 3h. Fig. 8D (from left to right): SPI treated with Endo-Stern Enzyme Bp, Protease Neutral, Flavourzyme and Alcalase.

[0079] Figs. 9A-9D. are images of PPI cheese melting test on pizza. Fig. 9A (from left to right): marketed vegan cheese, dairy cheese, native PPI. Fig. 9B (from left to right): PPI treated with PG for 0.5, 1.5 and 3h. Fig. 9C (from left to right): PPI treated with papain for 0.5, 1.5 and 3h. Fig. 9D (from left to right): PPI treated with Endo-Stern Enzyme Bp, Protease Neutral and Flavourzyme.

[0080] Figs. 10A-10D. are images of SPI cheese melting test on pizza. Fig. 10A (from left to right): marketed vegan cheese, dairy cheese, native SPI. Fig. 10B (from left to right): SPI treated with PG for 0.5, 1.5 and 3h. Fig. 10C (from left to right): SPI treated with papain for 0.5, 1.5 and 3h. Fig. 10D (from left to right): SPI treated with Endo-Stern Enzyme Bp, Protease Neutral and Flavourzyme.

[0081] Fig. 11. is a graph of stretchability test of PPI with PG and papain treatments in different DH%.

[0082] Fig. 12. is a graph of stretchability test of PPI with different enzyme treatments.

[0083] Fig. 13. is a graph of stretchability test of SPI with PG and papain treatments in different DH%.

[0084] Fig. 14. is a graph of stretchability test of SPI with different enzyme treatments.

[0085] Figs. 15A-15B are graphs presenting protein solubility profile of 20% wt PPI mixture in water after treatment with 15A. PG and 15B. papain.DETAILED DESCRIPTION

[0086] The cheese substitute (e.g. hard / semi-hard cheese substitute) of the present invention exhibits surprising and unexpected properties that overcome longstanding challenges in plant-based dairy alternatives. The inventors subjected various plant proteins to enzymatic pretreatment, so as to result in a modified protein with superior water solubility and characterized (i) a hydrolysis degree between about 3 and about 20% ; (ii) a molar ratio between glutamine and glutamic acid residues of below 1: 10 or both (i) and (ii). By combining the modified protein with polysaccharide (such as starch), the food product achieves meltability, stretchability, and texture that closely mimics dairy cheese, while maintaining desirable structural properties at room temperature.

[0087] Due to significantly improved water solubility of the modified protein of the invention, it is possible to incorporate a greater amount of protein into the cheese substitute compared toModified protein

[0088] In one aspect of the present invention there is provided a modified protein comprising a hydrolyzed plant protein characterized by: (i) a hydrolysis degree between 0.5 and 40% or between 0.5 and 30%; (ii) a molar ratio between glutamine and glutamic acid residues of below 1: 10 or both (i) and (ii). In some embodiments, the modified protein is characterized by at least one of: an alpha helix content of at least 50%; an average molecular weight below 120 kDa; and a water solubility above 25%, as determined based on total nitrogen content.

[0089] In some embodiments, the modified protein is obtained by contacting a plant protein with: (a) an enzyme having a deamidase and a proteolytic activity; or (b) a first enzyme having a deamidase activity and a second enzyme having a proteolytic activity.

[0090] As used herein, the term "deamidase" refers to an enzyme that catalyzes the removal of amide groups from amino acids in proteins, particularly the conversion of glutamine to glutamic acid residues. Deamidase activity may be associated with enzymes such asprotein glutaminase (PG), protein deamidase, and amidohydrolase.

[0091] In some embodiments, the enzyme having deamidase activity comprises amidohydrolase, protein deamidase, protein glutaminase, including any structural homologs and any combination thereof.

[0092] In some embodiments, the enzyme having proteolytic activity comprises any one of: pepsin, trypsin, chymotrypsin, papain, endoprotease, exoprotease, carboxypeptidase, aminopeptidase, or any combination thereof.

[0093] As used herein, the term "enzyme having a proteolytic activity" refers to a protease that catalyzes the hydrolysis of peptide bonds within protein molecules, typically breaking them down into smaller peptides. This enzyme class includes endoproteases that cleave internal peptide bonds within protein chains. Examples of proteases with proteolytic activity include, but are not limited to, pepsin, trypsin, chymotrypsin, papain, and various bacterial or fungal proteases.

[0094] In some embodiments, the protease with proteolytic activity is devoid of exopeptidase activity, i.e. enzymatic activity comprising cleavage of amino acids from the ends of protein chains.

[0095] In some embodiments, the plant protein may be derived from any plant source. The plant source may include, but is not limited to, legumes, grains, seeds, nuts, vegetables, or any combination thereof. Examples of specific plant sources may include pea, soy, chickpea, fava bean, lentil, quinoa, potato, garbanzo, yellow pea, rice, rye, golden lentil, chana dal, sorghum, sprouted green lentil, white lima bean, hemp, corn, rapeseed, canola, or any combination thereof. In some embodiments, the plant protein is derived from a vegetable protein. The vegetable protein may be obtained from various vegetable sources, including but not limited to leafy greens, root vegetables, cruciferous vegetables, squash, or any combination thereof.

[0096] Examples of specific vegetable sources may include pea, soy, chickpea, fava bean, lentil, quinoa, potato, garbanzo, yellow pea, rice, rye, golden lentil, chana dal, sorghum, sprouted green lentil, white lima bean, hemp, corn, rapeseed, canola, or any combination thereof.

[0097] The term "derived" encompasses inter alia a protein obtained by isolation from plant matter or biomass. This may include processes such as extraction, purification, fractionation, or concentration of proteins from plant sources. In some cases, the derivationprocess may involve mechanical methods like grinding, milling, or pressing of plant materials. Chemical extraction using solvents, acids, or bases may also be employed. Separation techniques such as centrifugation, filtration, or membrane separation may be used to isolate the proteins. The derivation process may optionally include additional steps like defatting, protein precipitation, drying, or spray drying to obtain protein isolates or concentrates. In some instances, enzymatic treatments or fermentation processes may be applied to modify or enhance the properties of the derived plant proteins.

[0098] In some embodiments, the modified protein has a hydrolysis degree between 2 and 30%. In some embodiments, the modified protein has a hydrolysis degree between 0.5 and 40%. In some embodiments, the modified protein has a hydrolysis degree between 0.5 and 30%.

[0099] As used herein, the term "hydrolysis degree" refers to the extent of protein hydrolysis, measured using the o-phthalaldehyde (OPA) method. The hydrolysis degree represents the percentage of peptide bonds cleaved during the enzymatic treatment of the protein.

[0100] In some embodiments, the hydrolysis degree of the modified protein is between 2 and 30%. In some embodiments, the hydrolysis degree is between 0.5 and 40%, between 0.5 and 30%, between 0.5 and 25%, between 0.5 and 20%, between 1 and 40%, between 1 and 30%, between 1 and 25%, between 1 and 20%, between 2 and 40%, between 2 and 30%, between 2 and 25%, between 2 and 5%, between 2 and 20%, between 3 and 20%, between 3 and 15%, between 3 and 17%, between 3 and 18%, between 3 and 25%, between 4 and 20%, between 4 and 25%, between 5 and 20%, between 5 and 15%, between 5 and 25%, between 5 and 30%, between 5 and 10%, between 10 and 15%, between 15 and 20%, between 20 and 25%, or between 25 and 30% including any range between.

[0101] In some embodiments, the molar ratio between glutamine and glutamic acid residues within the modified protein is reduced as compared to pristine protein. In some embodiments, the reduced molar ratio is below 1:10, or between 1:2 and 1: 10, between 1:2 and 1:20, between 1:2 and 1:3, between 1:3 and 1:4, between 1:4 and 1:5, between 1:5 and 1:6, between 1:5 and 1:10, between 1:5 and 1:8, between 1:5 and 1:9, between 1:5 and 1: 15, between 1:5 and 1:20, between 1:6 and 1:7, between 1:7 and 1:8, between 1:8 and 1:9, or between 1:9 and 1: 10. including any range between.

[0102] In some embodiments, the reduced molar ratio is reduced by at least 20%, at least 50%, at least 100%, at least 500%, at least 1000%, between 0.1% and 1000%, between 0.1% and 10%, between 5 and 50%, between 10 and 100%, between 50 and 250%, between 100 and 500%, between 250% and 750%, between 750 and 1000%, including any range in between, In some embodiments, the reduced molar ratio is determined by a C=O peak ratio between an amide and a carbonyl ester, obtained from an FTIR spectrum.

[0103] In some embodiments, the modified protein has at least 15%, at least 30% at least 50%, at least 100%, at least 150%, or between 30 and 150% greater water solubility compared to the native protein, as determined based on total nitrogen content.

[0104] In some embodiments, the modified protein is in a powder form. In some embodiments, the powder is a dry powder. In some embodiments, the dry powder has a water content of at most 10%, at most 5%, at most 2%, or at most 0.5.

[0105] In some embodiments, the water content of the dry powder is between 0.01 and 10%, between 0.01 and 0.1%, between 0.1 and 1%, between 1 and 2%, between 1 and 5%, between 3 and 7%, or between 5 and 10%.

[0106] In some embodiments, the modified protein is packaged in a sealed packing. In some embodiments, the sealed packing is a gas-tight packing. In some embodiments, the sealed packing may include plastic packaging, foil packaging, airtight packaging, moisture- tight packaging, or aseptic packaging. In some embodiments, the sealed packing helps maintain the stability / shelf-life and quality of the modified protein during storage and transportation.

[0107] In some embodiments, the modified protein is a texture forming additive. In some embodiments, the modified protein is characterized by gel forming properties when in contact with water.

[0108] According to one aspect there is provided a food product comprising (i) a modified protein, (ii) a polysaccharide and (iii) an oil; wherein: the (i) and the (ii) are non-crosslinked; the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 5 and 40%; and optionally is further characterized by a molar ratio between glutamine and glutamic acid residues of below 1:10; the food product is a homogenous solid at a temperature below a meltability onset point, and the meltability onset point is at a temperature ranging between 30 and 40°C.

[0109] In some embodiments, the food product further comprises trace amount of protease, protein deamidase, amidohydrolase or any combination thereof. In some embodiments, the trace amounts encompass at most 20,000 ppm, 10,000 ppm, 5000 ppm, 1000 ppm, 250 ppm, 100 ppm, between 10 and 10000 ppm, between 10 and 5000 ppm, between 10 and 100 ppm, including any range in between.

[0110] In some embodiments, the food product of the invention is a food grade product consisting or consisting essentially of food grade constituents. In some embodiments, the food product of the invention is a food grade product consisting or consisting essentially of food grade plant constituents, i.e. the food product of the invention is devoid of animal-based constituents or comprises up to 5%, up to 1%, up to 0.5%, up to 0.1%, up to 0.01 %w / w of animal based constituents, by dry weight of the food product.

[0111] As used herein, the term “food grade” refers to a product consisting of food-grade ingredients, generally recognized as safe and approved for human consumption by a corresponding regulatory authority (i.e., GRAS). The concentration of each of the constituents within the food grade doesn’t exceed a toxicity limit for the specific constituent as determined by the corresponding regulatory authority.

[0112] In some embodiments, amidohydrolase further encompasses an enzyme having amidohydrolase activity. Non limiting examples for the amidohydrolase are beta-lactamase, histone deacetylase, asparaginase, urease and glutaminase, including any structural homolog, and any combination thereof. In some embodiments, amidohydrolase is glutaminase. In some embodiments, amidohydrolase is protein glutaminase. In some embodiments, amidohydrolase is glutaminase. In some embodiments, amidohydrolase is protein deamidase. As used herein the term “homolog” refers to an enzyme with at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 99%, or 100% sequence homology to any of the enzymes disclosed herein, including any range between. Such enzymes can be used in their free form or immobilized on polymeric beads in order to facilitate their recovery in the process.

[0113] The terms “homology” or “identity”, as used interchangeably herein, refer to sequence identity between two amino acid sequences or two nucleic acid sequences, with identity being a stricter comparison. The phrases “percent identity or homology” and “% identity or homology” refer to the percentage of sequence identity found in a comparison oftwo or more amino acid sequences or nucleic acid sequences. Two or more sequences can be anywhere from 0-100% identical, or any value there between. Identity can be determined by comparing a position in each sequence that can be aligned for purposes of comparison to a reference sequence. When a position in the compared sequence is occupied by the same nucleotide base or amino acid, then the molecules are identical at that position. The degree of identity of amino acid sequences is a function of the number of identical amino acids at positions shared by the amino acid sequences. A degree of identity between nucleic acid sequences is a function of the number of identical or matching nucleotides at positions shared by the nucleic acid sequences. A degree of homology of amino acid sequences is a function of the number of amino acids at positions shared by the polypeptide sequences.

[0114] The following is a non-limiting example for calculating homology or sequence identity between two sequences (the terms are used interchangeably herein). The sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non- homologous sequences can be disregarded for comparison purposes). The optimal alignment is determined as the best score using the GAP program in the GCG software package with a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frame shift gap penalty of 5. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The identity percentage between the two sequences is a function of the number of identical positions shared by the sequences.

[0115] In some embodiments, % homology or identity as described herein are calculated or determined using the basic local alignment search tool (BLAST). In some embodiments, % homology or identity as described herein are calculated or determined using Blossum 62 scoring matrix.

[0116] In some embodiments, the modified protein is substantially non-crosslinked. In some embodiments, the modified protein and the polysaccharide are substantially noncrosslinked. In some embodiments, the non-crosslinked modified protein contains less than1% of crosslinked tyrosine residues, relative to the total amount of tyrosine residues (e.g. as determined by NMR, FTIR, or RAMAN).

[0117] In some embodiments, the protein is a non-animal protein. In some embodiments, the modified protein is from an animal origin (also referred to as animal modified protein). In some embodiments, the modified protein comprises both a non-animal protein and an animal protein. In some embodiments, the animal protein is selected from: dairy protein. In some embodiments, the non-animal protein is selected from: fruit modified protein, vegetable modified protein, seed modified protein, algal modified protein, fungal modified protein, yeast modified protein, legume modified protein, or any combination thereof. In some embodiments, the modified protein is substantially devoid of an animal protein.

[0118] In some embodiments, the non-animal protein is a plant protein. In some embodiment, the plant protein is selected from: pea, soybean, chickpea, quinoa, potato, garbanzo, fava beans, yellow pea, rice, rye, golden lentil, chana dal, sorghum, sprouted green lentil, white lima bean, hemp, corn, rapeseed, canola, including any fraction (or hydrolysate), derivative, or any combination thereof.

[0119] In some embodiment, the modified protein utilized in the instant invention is a food grade modified protein. In some embodiment, the modified protein comprises a single protein species, or a plurality of chemically distinct proteins. In some embodiment, the modified protein is a natural product. In some embodiment, the modified protein is derived from a natural product. In some embodiments, the term “derived from” encompasses any industrial processing such as purification, isolation, fractionation, chemical modification, etc. In some embodiments, the modified protein is a modified protein hydrolysate. In some embodiment, the modified protein is characterized by a chemical purity of between 50 and 99.9%, between 50 and 70%, between 70 and 80%, between 80 and 85%, between 85 and 90%, between 90 and 95%, between 95 and 99.9%, including any range in between.

[0120] In some embodiments, the modified protein is derived from a natural product, or from a natural protein isolate, and further comprises impurities such as polysaccharides and oligosaccharides. In some embodiments the modified protein comprises non-protein impurities up to 90%, up to 80%, up to 60%, up to 60%, up to 40%, up to 30%, up to 20%, up to 10% and between 1 and 90%, between 1 and 50%, between 10 and 40%, between 10and 20%, between 15 and 20%, between 20 and 30%, between 30 and 40%w / w of the modified protein material, including any range in between.

[0121] In some embodiments, the modified protein is obtained by treatment of a natural protein (e.g. natural plant protein, or plant protein isolate) with an enzyme having a deamidase activity (e.g. amidohydrolase; deamidase, protein glutaminase, etc.): and wherein the modified protein is characterized by a reduced ratio between Gln:Glu and / or AsmAsp residues.

[0122] In some embodiments, the food product of the invention is characterized by a reduced molar ratio between glutamine and glutamic acid residues (and / or between asparagine and aspartic acid residues) within the modified protein as determined by FTIR, wherein reduced is by at least 10%, at least 100%, at least 500%, at least 1000%, between 0.1% and 1000%, between 0.1% and 10%, between 5 and 50%, between 10 and 100%, between 50 and 250%, between 100 and 500%, between 250% and 750%, between 750 and 1000%, including any range in between, as compared to pristine protein. In some embodiments, the reduced molar ratio is determined by a C=O peak ratio between an amide and a carbonyl ester, obtained from an FTIR spectrum. Exemplary determination of the C=O peak ratio between the amide and the carbonyl ester peak ratio is as described in Fig. 1 A and Fig. IB.

[0123] In some embodiments, the transformation degree of glutamine to glutamic acid residue (and / or Asn to Asp) within the modified protein is between about 1 to 95%, between 1 and 5%, between 3 and 7%, between 5 and 10%, between 10 and 15%, between 15 and 25%, between 25 and 35%, between 35 and 45%, between 45 and 55%, between 55 and 65%, between 65 and 75%, between 75 and 85%, between 85 and 95%, including any range in between. The transformation degree was measured according to Church FC, Porter DH, Catignani GL, Swaisgood HE. An o-phthalaldehyde spectrophotometric assay for modified proteinases. Anal Biochem. 1985 May l;146(2):343-8. Exemplary transformation degree graph is described in Fig. 2.

[0124] In some embodiments, the modified protein is a hydrolyzed protein.

[0125] In some embodiments, the hydrolyzed protein is characterized by a degree of hydrolysis of between about 5 to 40%, between 5 and 30%, between 10 and 25%, between 10 and 30%, between 1 and 5%, between 5 and 10%, between 10 and 15%, between 15 and20%, between 20 and 25%, between 25 and 30%, between 30 and 40%, including any range in between. The degree of hydrolysis was measured using the o-phthalaldehyde (OPA) method.

[0126] In some embodiments, the hydrolyzed protein is characterized by an alpha helix content of at least 50%, of at least 60%, of at least 70%, and between 50 and 100%, between 50 and 60%, between 60 and 70%, between 70 and 80%, between 80 and 90%, between 90 and 100%, including any range in between. As used herein alpha helix content refers to a ratio between alpha helix and one or more secondary structures selected from beta sheet and random structure. Alpha helix content can be measured by circular dichroism or RAMAN, FTIR, NMR, etc.

[0127] In some embodiments, the hydrolyzed protein is a protein hydrolysate comprising a plurality of peptides. In some embodiments, the plurality of peptides is a mixture of peptides characterized by a distinct sequence, and / or distinct molecular weight. In some embodiments, the plurality of peptides is characterized by an alpha helix content of at least 50%, of at least 60%, of at least 70%, and between 50 and 100%, between 50 and 60%, between 60 and 70%, between 70 and 80%, between 80 and 90%, between 90 and 100%, including any range in between. In some embodiments, the plurality of peptides comprises a chemically distinct peptides (e.g., peptides having different amino acid sequence, and / or different molecular weight).

[0128] In some embodiments, the plurality of peptides is characterized by an average MW below 120kDa, below 80kDa, below 60kDa, between 1 and 100 kDa, between 1 and 80 kDa, between 1 and 60 kDa, between 1 and 50 kDa, between 1 and 5 kDa, between 5 and 10 kDa, between 5 and 20 kDa, between 5 and 40 kDa, between 10 and 15 kDa, between 15 and 20 kDa, between 20 and 25 kDa, between 20 and 30 kDa, between 30 and 40 kDa, between 40 and 50 kDa, between 40 and 80 kDa, between 40 and 60 kDa, between 30 and 80 kDa, including any range in between. In some embodiments, the plurality of peptides is characterized by an MW distribution (PDI) of between 0.2 and 1, including any range between. In some embodiments, the plurality of peptides is characterized by an average MW below 100 or below 80kDa, and by alpha helix content of above 50%.

[0129] As used herein, the terms "molecular weight" (MW) and “average MW” are used herein interchangeably and refer to weight average MW. For proteins / peptides, molecular weight may be determined using various analytical techniques, including but not limited to:

[0130] 1. Gel electrophoresis: Proteins can be separated based on their size using techniques such as sodium dodecyl sulfate -polyacrylamide gel electrophoresis (SDS- PAGE), with molecular weight estimated by comparison to known protein standards.

[0131] 2. Mass spectrometry: Techniques like matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) or electrospray ionization (ESI) mass spectrometry can provide accurate molecular weight measurements of proteins.

[0132] 3. Size exclusion chromatography: This method separates proteins based on their size as they pass through a column, with molecular weight estimated by comparison to known standards.

[0133] 4. Ultracentrifugation: Analytical ultracentrifugation can be used to determine the molecular weight of proteins based on their sedimentation behavior.

[0134] 5. Light scattering: Techniques such as dynamic light scattering (DLS) or multiangle light scattering (MALS) can be used to estimate the molecular weight of proteins in solution. As used herein, the average MW is determined by SEC-MALS.

[0135] The polydispersity index (PDI) may be used to describe the breadth of the molecular weight distribution in a protein sample. PDI is calculated as the ratio of the weight-average molecular weight (Mw) to the number-average molecular weight (Mn) of a polymer or protein sample:PDI=Mw / Mn, where:Mw (weight-average molecular weight) is calculated as:Mw=^iNiMi2 / ^iNiMiMn (number-average molecular weight) is calculated as:Mn=£iNiMi / £iNi

[0136] In these equations: Ni is the number of molecules of molecular weight Mi; and i represents different molecular weight species in the sample.

[0137] In some embodiments, the modified protein is obtained by treatment of a natural protein (e.g. natural plant protein, or plant protein isolate) with a first enzyme having deamidase activity and by a second enzyme having proteolytic activity, wherein the modified peptide is a hydrolyzed peptide characterized by a degree of hydrolysis, average MW and optionally by alpha helix content, as disclosed above; and wherein the modified peptide is further characterized by a reduced molar ratio between glutamine and glutamic acid residues (and / or between asparagine and aspartic acid residues) as disclosed above.

[0138] In some embodiments, the modified protein is obtained by enzymatic hydrolysis of a protein. In some embodiments, enzymatic hydrolysis comprises hydrolysis via a proteolytic enzyme, such as exoprotases, endproteases, protein deamidases, or any combinations thereof.

[0139] Non limiting examples of the proteolytic enzyme are pepsin, trypsin, chymotrypsin, papain, endoprotease, exoprotease, carboxypeptidase, protein deamidase or aminopeptidase including any combination thereof.

[0140] In some embodiments, the proteolytic enzyme is or comprises papain.

[0141] In some embodiments, dry weight percentage of the modified protein within the food product (i.e. non-hydrated food product) ranges between 2 and 60%, between 10 and 60%, between 20 and 60%, between 30 and 60%, between 20 and 40%, between 20 and 50%, between 20 and 30%, between 25 and 35%, between 30 and 40%, including any range or value therebetween.

[0142] In some embodiments, a weight concentration of the fat in the non-hydrated food product is between 5 and 50%, between 5 and 40%, between 10 and 50%, between 10 and 30%, between 10 and 20%, including any range or value therebetween.

[0143] In some embodiments, a weight ratio between the modified protein and the polysaccharide with the food product of the invention is between 1 : 10 and 10: 1, between 1 :5 and 5:1, between 1:3 and 3: 1, between 3:1 and 1: 1, between 2: 1 and 1: 1, between 1:1 and 1:2, between 1: 1 and 1:3, including any range in between.

[0144] In some embodiments, the food product is a homogeneous solid. In some embodiments, the homogeneous solid comprises the entire constituents uniformly distributed therewithin, and wherein the homogenous solid is devoid of phase separation (e.g. oil / solid constituents separation). In some embodiments, the homogeneous solidmaintains its physico-chemical properties (e.g., glass transition temperature, melting onset, Young’s modulus, elongation) within the entire dimensions (lengths and width dimensions) thereof. In some embodiments, the food product is a homogenous solid (i.e. is devoid of a visible phase separation, and substantially retains its shape) at a temperature below the melting onset point. In some embodiments, the food product is a homogenous solid (i.e. is devoid of a visible phase separation, and substantially retains its shape) at a temperature below 60°C, below 50°C, below 45°C, below 40°C, below 39°C, below 38°C and between 20 and 60°C, between 20 and 50°C, between 20 and 55°C, between 20 and 45°C, between 20 and 40°C, between 20 and 38°C, between 20 and 39°C, including any range in between.

[0145] In some embodiments, the protein molecules are substantially devoid of clusters or precipitations within the homogenous solid.

[0146] The term “fat” as used herein encompasses (plant) oil (e.g. a liquid oil at a temperature of about 25C), (plant) fat with different solid fat content (SFC), or both. The term “fat” as used herein encompasses natural plant oil / fat, and further encompasses modified plant oil / fat, such as hydrogenated plant oil.

[0147] In some embodiments, the fat has a melting point below 40C, below 38C, below 35C, below 30C, below 25C, below 20C, below 10C, including any range between. In some embodiments, the fat is or comprises a mixture of fat and oil. In some embodiments, at least one of the fat constituents is characterized by a melting point between 10 and 25C. In some embodiments, a first fat constituent is characterized by a melting point between 10 and 25C and a second fat constituent is characterized by a melting point below 10C.

[0148] In some embodiments, the plant oil is a liquid oil (or a natural oil) comprising or consisting essentially of a saturated and / or unsaturated fatty acid; fatty acid mono-, di- and / or tri-glyceride; a phospholipid, a fatty acid phosphate ester (e.g. mono- or di-ester), or any combination thereof.

[0149] In some embodiments, the fat is an edible fat. In some embodiments, the fat is suitable for human consumption. In some embodiments, the fat is or comprises at least one plant fat, at least one plant oil or a combination thereof.

[0150] In some embodiments, the plant oil is selected from or comprises olive oil, sunflower oil, safflower oil, canola oil, linseed oil, macadamia oil, walnut oil, argan oil, bifractionated palm oil, refined palm oil, castor oil, rice oil, peanut oil, corn oil, sesame seedoil, soy bean oil, grapeseed oil, avocado oil, hemp seed oil, essential oil, including any combination thereof. Plant oil maybe partially hydrogenated.

[0151] In some embodiments, the plant fat is selected from or comprises Coconut Oil, Cocoa Butter, Palm Oil, Shea Butter, Mango Butter, including any combination thereof.

[0152] In some embodiments, the polysaccharide is a food grade constituent, wherein the polysaccharide comprises a modified starch or a native starch, or any combination thereof.

[0153] In some embodiments, the polysaccharide (further encompassing an oligosaccharide) is a hydrocolloid polysaccharide such as starch (modified or non-modified starch), cellulose, alkylated cellulose, hydroxyalkylated cellulose, carboxylated cellulose, xanthan, guar gum, locust bean gum, gum arabic, pectin, carrageenan, alginate, gellan and agar, including any salt or any combination thereof.

[0154] In some embodiments, the polysaccharide is a plant starch (modified or nonmodified). In some embodiments, the plant starch comprises potato, corn (maize), rice, tapioca starch, or any combination thereof.

[0155] In some embodiments, the food product further comprises water, wherein the w / w percentage of water within the food product of the invention is between 0.1 and 80% w / w, between 0.1 and 5%, between 0.1 and 10%, between 5 and 25%, between 10 and 20%, between 25 and 50%, between 20 and 30%, between 30 and 40%, between 40 and 50%, between 50 and 60%, between 60 and 70%, between 70 and 80%, including any range in between. In some embodiments, the water is bonded by physical interactions (e.g., hydrogen bonding, dipole-dipole interactions, electrostatic interactions, etc.) to the plant protein, and / or to the polysaccharide.

[0156] In some embodiments, the food product consists essentially of the modified protein, water, fat, and the polysaccharide. In some embodiments, the food product consists essentially of the modified protein, water, fat, and the polysaccharide, wherein the modified protein and the polysaccharide have a synergetic effect (i.e. provide a predetermined texture, organoleptic and physico-mechanical properties such as elasticity, hardness, gumminess to the food product). In some embodiments, at least 70%, at least 80%, at least 90%, at least 95% by weight of the food product consists of the modified protein, water and the polysaccharide. In some embodiments, the modified protein, water and the polysaccharide constitute between 70 and 95%, between 70 and 75%, between 75 and 80%, between 80 and85%, between 85 and 90%, between 90 and 95% by weight of the food product, including any range in between.

[0157] In some embodiments, the food product is a hydrated-solid (is also referred to herein as “moist food product”). In some embodiments, the moist food product comprises water, wherein the water is bonded by physical interactions (e.g., hydrogen bonding, dipoledipole interactions, electrostatic interactions, etc.) to the plant protein, and / or to the polysaccharide. In some embodiments, the food product is a hydrogel.

[0158] In some embodiments, the w / w percentage of the water within the moist food product of the invention is between 5 and 70% w / w, between 5 and 15%, between 10 and 20%, between 20 and 30%, between 30 and 40%, between 40 and 50%, between 50 and 60%, between 60 and 70% including any range in between. In some embodiments, the food product further comprises the first enzyme having a deamidase activity and / or the second enzyme having a proteolytic activity. In some embodiments, a concentration of the first enzyme and / or second enzyme within the food product is between 1 and 20000 ppm, between 10 and 20000 ppm, between 1 and 1000 ppm, between 10 and 1000 ppm, including any range in between.

[0159] In some embodiments, the food product is a dehydrated- solid (is also referred to herein as “dry food product”), comprises at most 10%, at most 5% water, at most 2, at most 0.5% water, between 0.01 and 10%, between 0.01 and 0.1%, between 0.1 and 1%, between 1 and 2%, between 1 and 5%, between 3 and 7%, between 5 and 10%, including any range in between.

[0160] In some embodiments, the dry food product consists essentially of the porous matrix formed by the network of modified protein and the polysaccharide of the invention. In some embodiments, the dry food product is a solid, or a porous solid matrix (e.g., a sponge). In some embodiments, the dry food product is a powder. In some embodiments. The dry product is lyophilized.

[0161] In some embodiments, the dry food product is a porous matrix. In some embodiments, the porous matrix is characterized by a porosity of between 40 and 90%, between 40 and 50%, between 45 and 55%, between 50 and 60%, between 60 and 70%, between 70 and 80%, between 80 and 90%, including any range in between.

[0162] In some embodiments, the dry food product is a low-density matrix. In some embodiments, the dry food product is characterized by a density of less than 1.0 g / cm3, less than 0.98 g / cm3, less than 0.95 g / cm3, less than 0.93 g / cm3, and between 0.85 and 1.0 g / cm3, between 0.85 and 0.88 g / cm3, between 0.88 and 0.91 g / cm3, between 0.91 and 0.93 g / cm3, between 0.93 and 0.95 g / cm3, between 0.95 and 0.97g / cm3, and between 0.97 and 0.99 g / cm3, between 0.99 and 1.0 g / cm3including any range in between.

[0163] In some embodiments, the dry food product has at least one dimension; a length, width, height or depth of between 1 cm and 1 m, between 1 cm and 0.1m, between 1 and 25 cm, between 10 and 50 cm between 0.1 and 0.2m. between 0.2 and 0.3m, between 0.3 and 0.4m, between 0.4 and 0.5m, between 0.5 and 0.6m, between 0.6 and 0.7m, between 0.7 and 0.8m, between 0.8 and 0.9m, between 0.9 and Im including any range in between.

[0164] As used herein, the term “matrix” refers to one or more porous continuous layers of networked chains, wherein each chain comprises the modified protein / peptide, and / or the polysaccharide. In some embodiments, the modified protein / peptide chains are randomly, and / or under certain order or control, distributed therewithin. Matrix may further include any materials incorporated within and / or interposed between the layers or within the pores. In some embodiments, the matrix comprises randomly oriented chains. In some embodiments, each chain within the matrix is in contact with at least one additional chain. In some embodiments, the chains are randomly distributed within the matrix, to obtain a three-dimensional mesh structure comprising a void space between the chains. In some embodiments, the chains are randomly distributed within the matrix thus forming a plurality of pores (or void space). In some embodiments, the matrix comprises the plant polymer, as described herein. In some embodiments, the matrix comprises the polysaccharide. In some embodiments, the chains are randomly distributed within the matrix. In some embodiments, the matrix is substantially devoid of aligned or oriented chains. In some embodiments, the matrix is substantially devoid of chains aligned or oriented in a specific direction.

[0165] The term “layer” refers to a substantially homogeneous substance of substantially uniform-thickness which maintains its physico-chemical properties (e.g., glass transition temperature, Young’s modulus, elongation) with the entire dimensions (lengths and width dimensions) thereof. In some embodiments, each layer has a different physical structure and / or a different chemical composition. In some embodiments, each layer has the samephysical structure and / or the same chemical composition. In some embodiments, the term "layer", refers to a polymeric layer.

[0166] In some embodiments, the food product further comprises dietary supplements such as oil or fat with different solid fat content (SFC), unsaturated fatty acids in their ethyl ester, glyceride and phospholipid forms (such as Omega 3), aroma, flavors, vitamins, coloring agents, preservatives, but not limited to. In some embodiments, the dietary supplements weight concentration within the food product is up to 30% and between 0.1 and 30%, between 0.1 and 0.5%, between 0.5 and 1%, between 1 and 2%, between 2 and 3%, between 3 and 4%, between 4 and 5%, between 5 and 6%, between 6 and 7%, between 7 and 8%, between 8 and 9%, between 9 and 10%, between 10 and 20%, between 15 and 25%, between 20 and 30%, including any range in between.

[0167] In some embodiments, the food product of the invention, is shapeable, spreadable or cut able, shred-able. In some embodiments, the food product of the invention is characterized by a predetermined shape. In some embodiments, the predetermined shape is any of a sphere, a hemisphere, a hollow sphere, a cylinder, a hollow cylinder, a hollow hemisphere, a cone, a pyramid, a horseshoe, or any other 3-D shape. In some embodiments, the predetermined shape is an irregular shape. The food product of the invention can be generally shaped as a sphere, incomplete-sphere, a rod, a cylinder, a ribbon, a sponge, and any other shape, or can be in a form of a cluster of any of these shapes or can comprise a mixture of one or more shapes.

[0168] In some embodiments, non-limiting examples of shaped food products are sphere, disc-like shape, chunk (cube), cylinder, slices, strips, etc.

[0169] In some embodiments, the food product is meltable at a temperature above the melting onset point. In some embodiments, meltability comprises a diameter increase between 170 and 350%, between 170 and 400%, between 170 and 320%, between 170 and 310%, between 170 and 300%, between 180 and 250%, between 180 and 220%, between 180 and 200%, as determined by Schreiber test.

[0170] In some embodiments, the food product undergoes melting under heating at a temperature up to 350°C, and between 40 and 200°C, between 40 and 60°C, between 40 and 100°C, between 40 and 150°C, including any range in between. In some embodiments, the food product is characterized by a melting onset point between 30 and 45 °C, between 30 and40°C, between 32 and 45 °C, between 35 and 45 °C, between 35 and 43 °C, between 38 and 45 °C, between 38 and 43 °C, between 38 and 42°C, including any range between.

[0171] In some embodiments, the food product is shapeable, spreadable or cut-able, shredable; and wherein the food product maintains its shape under heating at a temperature up to 40°C.

[0172] In some embodiments, the food product is contained in a suitable package, such as plastic packaging, foil packaging, airtight packaging, moist tight packaging, aseptic packaging, but not limited to.

[0173] In some embodiments the food product is selected from: raw, fried, baked, vacuum, boiled, cooked, poached, but not limited to.

[0174] In some embodiments, the food product is vegetarian food product. In some embodiments, the food is a vegan-based food product. In some embodiments, the food product is devoid of animal product or any component of an animal product. In some embodiments, the food product is a dairy food product substitute.

[0175] In some embodiments, the dairy food product comprises any one of;

[0176] In some embodiments, the food product is a non-dairy cheese substitute. In some embodiments, the food product is a semi-hard cheese substitute.

[0177] In some embodiments, the food product of the invention is characterized by at least one of: (i) a hardness between 5 and 40N, (ii) adhesiveness between 0.1 and 3Nmm, and (iii) a fracture between 5 and 30N, or by any combination of (i)-(iii), as determined by Texture Profile Analyzer.Method 1

[0178] In another aspect, there is provided a method for manufacturing the food product of the invention, the method comprises (i) contacting a protein with (a) an enzyme having a deamidase and a proteolytic activity; or (b) a first enzyme having a deamidase activity and with a second enzyme having a proteolytic activity, thereby obtaining the modified protein characterized by a molar ratio between glutamine and glutamic acid (and / or Asn: Asp) of at most 1 : 10, or at most 1:2, (ii) mixing the modified protein with the polysaccharide and fat, thereby obtaining the food product.

[0179] In some embodiments, the modified protein is a hydrolyzed protein, characterized by a molar ratio between glutamine and glutamic acid (and / or Asn:Asp) of at most 1: 10, and by at least one of: a hydrolysis degree between about 1 and about 30%, between about 1 and about 40% or between about 1 and about 20%; a molar ratio between glutamine and glutamic acid residues of below 1: 10; alpha helix content of at least 50%; average MW below 120kDa.

[0180] In some embodiments, step (i) comprises contacting the protein and (a) or (b) at a temperature between 10 and 70°C between 10 to 20°C, between 20 to 30°C, between 30 to 40°C, between 40 to 50°C, between 50 to 60°C, between 60 and 70°C, including any range in between. In some embodiments, the protein is suspended, dispersed, or solubilized in a water solution to obtain an aqueous composition comprising the protein. In some embodiments, the aqueous composition consists essentially of the protein. In some embodiments, the aqueous composition is devoid of starch.

[0181] In some embodiments, step (i) comprises mixing, shaking or stirring the aqueous composition. In some embodiments, step (i) is performed for a period of time sufficient for achieving the desired transformation degree (i.e. converting the glutamine into glutamic acid within the plant protein determined by Nessler, and / or the degree of hydrolysis determined by OP A method).

[0182] In some embodiments, the modified protein has at least 15%, at least 30% at least 50%, at least 100%, at least 150%, or between 30 and 150% greater water solubility compared to the native protein, as determined based on total nitrogen content. Total nitrogen of the protein can be converted to protein solubility by using a nitrogen / protein conversion factor. For pea protein the factor is 5.36 (average value).

[0183] In some embodiments, the modified protein has a water solubility above 25%, above 30%, above 35%, between 25 and 45%w / w, including any range between, as determined based on total nitrogen content.

[0184] The inventors observed that before performing the enzymatic modification (i.e. step i) the aqueous composition containing about 20%w / w of the plant protein in DDW was characterized by a paste-like appearance a was a non-flowable liquid (e.g. no flowability was detected upon shaking thereof using a standard laboratory shaker). After completion of step i, the resulting aqueous composition was a flowable liquid. This observation indicates a significantly improved water solubility / dispersibility of the modified plant protein, ascompared to pristine plant protein. Accordingly, the progression of step i of Method 1 or 2 can be monitored inter alia by the flowability of the aqueous composition (i.e. by determining viscosity, wherein viscosity at 20°C of below 100 or below 50 cP is indicative of a flowable liquid).

[0185] In some embodiments, step (i) comprises contacting the protein and the deamidase (such as protein deamidase, such as protein glutaminase, and / or modified protein hydrolase), separately or simultaneously at a temperature between 10 and 70°C between 10 to 20°C, between 20 to 30°C, between 30 to 40°C, between 40 to 50°C, between 50 to 60°C, between 60 and 70°C, including any range in between. In some embodiments, the protein is suspended, dispersed, or solubilized in a water solution to obtain an aqueous composition comprising the protein. In some embodiments, the aqueous composition consists essentially of the protein.

[0186] In some embodiments, step (i) comprises mixing, shaking or stirring the aqueous composition. In some embodiments, the aqueous composition is devoid of starch. In some embodiments, step (i) is performed for a period of time sufficient for achieving the desired transformation degree (converting the glutamine into glutamic acid within the plant protein) and optionally to arrive at a predetermined degree of hydrolysis.

[0187] In some embodiments, the protein is or comprises a plant protein, or a plant protein isolate, concentrate or precipitated wet plant protein, such as tofu. In some embodiments, the protein is or comprises a plant protein.

[0188] In some embodiments, the weight concentration of the protein in the aqueous composition is at least 1% and between 3 and 40%, between 3 and 30%, between 5 and 15%, between 15 and 25%, between 17 and 23%, between 25 and 35%, including any range in between. In some embodiments, the protein is a sole polymer in the aqueous composition.

[0189] In some embodiments, weight concentration of any one of (a) and (b) in the aqueous composition relative to the protein is at most 3%, and between 0.01 and 3%, between 0.01 and 0.05%, between 0.05 and 0.1%', between 0.1 and 0.5%, between 0.5 and 1 %, between 1 and 3%, including any range in between.

[0190] In some embodiments, the method further comprising performing a termination step thereby substantially arresting enzymatic activity of (a) or (b), wherein the termination step is performed prior to step (ii). In some embodiments, the termination step comprisesany of: heating the aqueous composition to a temperature of at least 90°C, (b) contacting with the aqueous composition with an inhibitor of (a) or (b), or exposing the aqueous composition to a pH below 4 or above 9.

[0191] In some embodiments, termination is performed after the desired transformation and / or hydrolysis degree was obtained.

[0192] In some embodiments, step (ii) comprises mixing the modified protein, the polysaccharide and fat at a temperature between 70 and 110 °C for a period of time sufficient for forming the food product. In some embodiments, the period of time sufficient for forming the food product is at least 5 min, at least 30 min, at least Ih, at least 2h, at least 3h, at least 4h, at least 8h, at least 16h, including any range in between.

[0193] In some embodiments, the mixing temperature is between 10 and 120°C between 10 to 20 °C, between 20 to 30 °C, between 20 and 110°C, between 30 to 40 °C, between 40 to 50 °C, between 50 to 60 °C, between 60 and 70°C, including any range in between.

[0194] In some embodiments, step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:2 and 10: 1, between 1:1 and 10: 1, between 1: 1 and 1:3, between 3: 1 and 1:3, between 1:1 and 1:2, between 1: 1 and 1:5, between 1:3 and 1:5, between 1:5 and 1:7, between 1:7 and 1:8, between 1:5 and 1:9, between 1:1 and 1:4, including any range in between.

[0195] In some embodiments, the method further comprises storage at a temperature between 4 and 8 °C for a period of time sufficient for forming stable food product. In some embodiments, the period of time sufficient for forming the food product is between 1 h to 10 days, at least Ih, at least 2h, at least 3h, at least 4h, at least 8h, at least 16h, at least 1 day, at least 2 days, at least 4 days, at least 8 days, at least 16 days including any range in between. In some embodiments, the storage temperature is between 0 and 4°C between 4 to 8 °C, including any range in between.

[0196] In some embodiments, the enzymes (a) or (b) Eire immobilized (i.e., bound to a solid support). In some embodiments, the immobilized enzyme is insoluble in water. In some embodiments, the enzyme is non-immobilized. In some embodiments, the enzyme is soluble in water.Method 2

[0197] In another aspect, there is provided a method for manufacturing the food product of the invention, the method comprises (i) contacting a protein with a proteolytic enzyme under conditions suitable for hydrolysis of the protein, thereby obtaining a modified protein; (ii) contacting the modified protein with polysaccharide and with fat under conditions appropriate for producing the food product. In some embodiments, the modified protein comprises a hydrolyzed plant protein (HPP). In some embodiments, the HPP is characterized by a hydrolysis degree between 3 and 40%, between about 1 and about 40%, between about 1 and about 30%, between about 1 and about 20%, between about 2 and about 20%, between about 2 and about 30%, between about 3 and about 30%, between about 3 and about 20%, between about 5 and about 30%, between about 5 and about 20%, including any range between.

[0198] In some embodiments, the HPP comprises a plurality of modified proteins (peptides) characterized by molecular weight (MW), by water solubility and by a secondary structure as disclosed hereinabove. In some embodiments, the conditions suitable for hydrolysis comprise mixing at a temperature between 10 and 70°C, between 10 to 20°C, between 20 to 30°C, between 30 to 40°C, between 40 to 50°C, between 50 to 60°C, between 60 and 70°C, including any range in between.

[0199] In some embodiments, the protein is suspended, dispersed, or solubilized in a water solution to obtain the aqueous composition. In some embodiments step (i) is performed for a period of time sufficient for obtaining the desired hydrolysis degree (determined by OPA).

[0200] In some embodiments, the weight concentration of the protein in the aqueous composition is as described in Method 1. In some embodiments, a weight concentration of the (proteolytic) enzyme(s) in the aqueous composition is as described in Method 1.

[0201] In some embodiments, the method further comprising performing a termination step thereby substantially arresting enzymatic activity of the proteolytic enzyme, wherein the termination step is performed prior to step (ii). In some embodiments, the termination step comprises any of: (a) heating at a temperature of at least 90°C, or (b) exposing to a pH below 4 or above 9.

[0202] In some embodiments, termination is performed after the desired degree of hydrolysis is achieved.

[0203] In some embodiments, step (ii) comprises contacting the HPP, the polysaccharide and fat at a temperature between 30 and 120°C for a period of time sufficient for forming the food product, wherein the ratios between the constituents are as described above.

[0204] In some embodiments, the period of time sufficient for forming the food product is at least 1 min, at least 2 min, at least 4 min, at least 8 min, at least 16 min including any range in between. In some embodiments, the mixing temperature is between 20 and 120°C, between 30 and 120°C, between 30 and 110°C, between 30 to 40°C, between 40 to 50°C, between 50 to 60°C, between 60 to 70°C, between 70 to 80°C, between 80 and 90°C, between 90 and 100°C, between 100 and 110°C, between 110 and 120°C including any range in between.

[0205] In some embodiments, after performing the termination step, step (i) of method 1 or method 2 further comprises isolating the modified protein from the aqueous composition. In some embodiments, isolation comprises separation of the modified protein from the aqueous solvent. In some embodiments, isolation is performed by any one of: drying the aqueous composition to obtain a powder comprising the modified protein; precipitation of the modified protein (e.g. by Salt Precipitation, pH adjustment to the isoelectric point (pl) of the modified protein, lowering the temperature of the peptide solution to induce precipitation, addition of non-solvents such as diethyl ether); or by separation (e.g. using liquid chromatography separation, SEC, centrifugation, etc. In some embodiments, drying comprises vacuum drying (e.g. lyophilization), heat drying, convection drying or spray drying.

[0206] In some embodiments, the proteolytic enzyme comprises any one of pepsin, trypsin, chymotrypsin, papain, endoprotease, exoprotease, carboxypeptidase, aminopeptidase, protein deamidase or any combination thereof. In some embodiments, the proteolytic enzyme comprises pepsin.

[0207] In some embodiments, the proteolytic enzyme is immobilized. In some embodiments, the proteolytic enzyme is insoluble in water. In some embodiments, the proteolytic enzyme is non-immobilized. In some embodiments, the proteolytic enzyme is soluble in water.

[0208] The food products disclosed herein can be characterized by organoleptic tests including hardness, stickiness, sandy texture, and binding. Test setup: a food product wascompressed by hand and cut with a knife to evaluate the hardness, stickiness to the knife, and visual assessment for sandy texture and bending.

[0209] The food products disclosed herein can be characterized by instruments such as texture profile analyzer (TPA, Lloyd, equipped with 100N load cell). Test setup: a food product with diameter of 2.3cm x 1.5cm (height) was compressed by 40% between two parallel plates at a speed of lOOmm / min, in two compression cycles, with a 0.1 second delay between cycles.

[0210] The food products disclosed herein can be characterized by pH meter to measure the acidity of the food products.

[0211] The food products disclosed herein can be characterized by shredding tool to evaluate the capability of shredding.

[0212] The food products disclosed herein can be characterized by melting test using modified Schreiber test or visual on pizza application. Test setup: a food product with diameter of 2.3cm x 1cm (height) placed in a Petri dish covered with the corresponding glass top then placed in oven preheated to 200°C for 20 min. The samples were removed and allowed to cool. Diameter of the spread at four different angles was measured. The measurement average was used to calculate the meltability by determining the percentage increase in diameter from the initial 23 mm. Melting on pizza application was visually assessed and scored with positive melting (1) or negative (0).General

[0213] As used herein the term “about” refers to ± 10%. Further, all numerical values, e.g., when referring the amounts or ranges of the elements constituting the formulation are approximations which are varied (+) or (-) by up to 10% of from the stated values. It is to be understood, even if not always explicitly stated that all numerical designations are preceded by the term "about".

[0214] The terms “comprises”, “comprising”, “includes”, “including”, “having” and their conjugates mean “including but not limited to”. Further, the term "comprises", "comprising", "includes", "including", “having” and their conjugates maybe alternatively used as “consisting”. For example the phrase “a composition comprises” also encompasses “a composition consists of’ or “a composition is”. As used herein, the term “comprises” including any grammatical form thereof may be used as “consisting essentially of’.

[0215] The term “consisting of means “including and limited to”. As used herein, the term “consisting essentially of’ with regards to the herein disclosed compositions refers to compositions including less than 2% w / w, less than 1% w / w, less than 0.5% w / w, less than 0.1% w / w, less than 0.05% w / w or less than 0.01% w / w of ingredients other than those disclosed. Each possibility is a separate embodiment.

[0216] The term “consisting essentially of’ means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure. Further, the term “consisting essentially of’ means that at least 80%, at least 85%, at least 90%, at least 92%, at least 95%, at least 97%, at least 99% or between 80 and 99%, between 90 and 99%, between 95 and 99%, between 80 and 100%, between 90 and 100%, between 95 and 100% by weight of the composition or article consists of the listed ingredients, including any range between.

[0217] The term “consisting of’ means “including and limited to”.

[0218] The word “exemplary” is used herein to mean “serving as an example, instance or illustration”. Any embodiment described as “exemplary” is not necessarily to be construed as preferred or advantageous over other embodiments and / or to exclude the incorporation of features from other embodiments.

[0219] The word “optionally” is used herein to mean “is provided in some embodiments and not provided in other embodiments”. Any particular embodiment of the invention may include a plurality of “optional” features unless such features conflict. The word “optionally” and the word "further" are used herein interchangeably.

[0001] As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

[0002] I some embodiments, the term “one or more” and the term “a” encompasses a single constituent, or a plurality (e.g., 2, 3, 4, 5 or 6) of constituents.

[0220] Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be consideredto have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0221] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases “ranging / ranges between” a first indicate number and a second indicate number and “ranging / ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

[0222] As used herein the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.

[0223] Other terms as used herein are meant to be defined by their well-known meanings in the art.

[0224] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub combination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

[0225] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.EXAMPLES

[0226] The inventor successfully utilized protease enzymes (e.g., protein glutaminase, protein deamidase, and papain and as an endoprotease) as a tool for hydrolyzing proteins obtaining a synergetic effect with hydrocolloids to yield the food product of the invention.EXAMPLE 1Preparation and characterization of the food product

[0227] Treatment of 10% w / w plant protein in water (pea protein isolate (PPI) or soy protein isolate (SPI), fava bean) with protein glutaminase (PG) or protein deamidase with endo protease. PPI, SPI or fava bean were mixed in distilled water containing 500 ppm sodium benzoate (inhibitor of microbial growth) and PG or protein deamidase with endo protease at a temperature between 40 and 60°C. The transformation degree of glutamine to glutamic acid in different reaction time intervals was examined, presented in Fig. 1 for PPI (determined by the Nesller). Endoprotease activity was examined and presented in Fig. 4 (determined by SDS and SEC MALS). In addition, the solubility of the modified protein was visually seen (Fig. 5A and Fig. 5B) and measured by total nitrogen content (Fig. 15A). At the end of the reaction the PG or protein deamidase with endo protease were inactivated by heat by immersing the reaction mixture container in a water bath and reach 90°C for 15min. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (10% wt. of the total wt.) with modified potato starch (17% wt. of the total wt.), EBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (55.9% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.8% wt. of the total wt.) were mixed in a speed of 2 until homogenized.Then, the dry mixture of modified protein, starch, EBG and salt was added to the thermomix, mixed at a speed of 4 for few minutes or until homogenized. Finally, melted oil (coconut and sunflower oil mixture, 15% wt. of the total wt.) was poured into the mixture gradually while mixing at speed 4 until homogenized. The mixture was heated to 85 °C for 5 minutes at speed 4 then poured hot into a mold and let it cool to RT then placed in refrigerator 4 to 8 °C for 7-10 days to obtain a shaped product (Fig. 6). Texture profile analysis (TPA, Floyd) of the food product was tested, along with pH, melting test (modified Schreiber test andcheese pizza melting test), stretchability in sandwich toast, shred-ability, and organoleptic tests. Results are summarized in Tables 1 - 8 and Figures 6 - 14.EXAMPLE 2Preparation and characterization of the food product

[0228] Organoleptic test and TPA present excellent hardness after treatment with PG or protein deamidase with endo protease activity for both pea and soy proteins that resemble dairy cheese (Table 1, 2 and 4). The results show that with increased DH%, the hardness of the product increases. In addition, the binding of semi-hard cheese is enhanced compared to native protein. The significant change in eliminating sandy texture from cheese formulations after protein incorporation is achieved after enzymatic treatment with PG or protein deamidase with endo protease as can be seen from the results (Fig. 6).

[0229] Shredding of cheese was applicable and presented clean cut edge, uniform shreds, and no clumping.

[0230] The modified Schreiber test presented diameter increase of dairy cheese to 228.78% ± 2.15, marketed vegan cheese of 190.01% ± 0.69 and native protein formulation of 145.02% ± 34.16 for pea and 217.5% ± 10.30 for soy protein (Table 7). PG or protein deamidase with endo protease treatment enhanced the melting in pea protein similarly to dairy cheese, while in soy protein it didn’t significantly affect the melting in this test but is similar to dairy cheese (Fig 7 and Fig. 8).

[0231] The melting cheese on pizza application is presented in Fig. 9 for pea and Fig. 10 for soy cheese. It can be seen that native protein formulation did not present good melting for pizza application, while the PG or protein deamidase with endo protease treatment proteins presented good melting.

[0232] The stretchability in sandwich toast presented an increase after PG or protein deamidase with endo protease treatment for soy protein compared to use of native protein. However, with pea protein it was decreased compared to native, on the other hand, the results show an increase of stretchability with increased DH% (Fig 11 - Fig.14). Furthermore, even at very low DH% (e.g. 0.5 and 1%) the food product is still meltable (as presented in Table 8), however in order to obtain a hard / semi-hard cheese substitute closely mimicking themeltability of the dairy cheese, it is preferable to use modified protein with DH% between 3 and 25% or between 5 and 25%.EXAMPLE 3Preparation and characterization of the food product

[0233] Treatment of 10% w / w plant protein in water PPI or SPI with papain. The method for preparation and cheese analog preparation is similar to example 1.

[0234] Organoleptic test and TPA present excellent hardness after papain treatment in both pea and soy proteins yielding plant-based cheese that resembles dairy cheese (Table 1, 2 and 4). The results show that increased DH% did not affect the hardness of the food product.

[0235] The binding of semi-hard cheese is enhanced compared to native protein in both pea and soy proteins. In addition, the significant change in eliminating sandy texture from cheese formulations after protein incorporation is achieved after enzymatic treatment with papain as can be seen from the results (Fig. 6).

[0236] However, sticky texture was presented in soy cheese analog after papain treatment.

[0237] Shredding of cheese was applicable and presented clean cut edge, uniform shreds, and no clumping.

[0238] The modified Schreiber test presented enhanced significancy the DI% increase after papain treatment in pea protein compared to native protein and is slightly higher than dairy cheese, while in soy protein it did not affect the melting in this test but is similar to dairy cheese. On the other hand, the melted cheese was dry compared to dairy and PG or protein deamidase with endo protease treatment (Fig 7 and Fig. 8).

[0239] The melting cheese on pizza application is presented in Fig. 9 for pea and Fig. 10 for soy cheese. It can be seen that native protein formulation did not present good melting for pizza application, while the papain treatment proteins presented good melting.

[0240] The stretchability in sandwich toast presented an increase with a factor of up to 2.4 after papain treatment with pea and soy proteins compared to native proteins. In addition, the results show an increase of stretchability with increased DH% in pea protein formulations (Fig 11 - Fig.14).EXAMPLE 4Preparation and characterization of the food product

[0241] Treatment of 10% w / w plant protein in water PPI or SPI with Endo-SternEnzyme Bp (SternEnzym). The method for preparation and cheese analog preparation is similar to example 1.

[0242] Organoleptic test and TPA present excellent hardness after Endo-SternEnzyme Bp treatment of pea protein. The final product has the shape and texture of dairy cheese (Table 1, 2 and 4). The binding of semi -hard cheese is enhanced compared to native protein in both pea and soy proteins. In addition, the significant change in eliminating sandy texture from cheese formulations after protein incorporation is achieved after enzymatic treatment with Endo-SternEnzyme Bp as can be seen from the results.

[0243] The modified Schreiber test presented enhanced DI% after treatment with Endo- SternEnzyme Bp for pea protein compared to native protein and similar to dairy cheese. While soy protein it didn’t affect the melting in this test (Fig 7 and Fig. 8).

[0244] The melting cheese on pizza application presented in Fig. 9 for pea and Fig. 10 for soy protein cheese shows that native soy protein formulation did not present good melting for pizza application, while the treated soy protein with Endo-SternEnzyme Bp treated proteins presented good melting.

[0245] The stretchability in sandwich toast presented an increase after Endo-SternEnzyme Bp treatment for pea and soy proteins compared to native protein (Fig 11 - Fig.14).EXAMPLE 5Preparation and characterization of the food product

[0246] Treatment of 10% w / w plant protein in water PPI or SPI with Protease Natural P (Sunson). The method for preparation and cheese analog preparation is similar to example 1.

[0247] Organoleptic test and TPA for cheese analog produced by treatment of pea protein with Protease Natural P resembles dairy cheese (Table 1, 2 and 4). The binding of semi-hard cheese is enhanced compared to native protein in both pea and soy proteins. In addition, the significant change in eliminating sandy texture from cheese formulations after proteinincorporation is achieved after enzymatic treatment with Protease Natural P as can be seen from the results.

[0248] The modified Schreiber test presented enhanced significancy the DI% increase after Protease Natural P treatment in pea protein compared to native and similar to dairy cheese, while in soy protein it did affect the melting in this test but is similar to dairy cheese (Fig 7 and Fig. 8).

[0249] The melting cheese on pizza application presented in Fig. 9 for pea and Fig 10 for soy cheese shows that native protein formulation did not present good melting for pizza application, while the Protease Natural P treated proteins presented good melting.

[0250] The stretchability in sandwich toast presented an increase after Protease Natural P treatment with pea and soy proteins compared to native protein (Fig 11 - Fig.14).EXAMPLE 6Preparation and characterization of the food product

[0251] Treatment of 10% w / w plant protein in water (pea protein isolate (PPI) or soy protein isolate (SPI)) with Flavourzyme (Novozymes). The method for preparation and cheese analog preparation is similar to example 1.

[0252] Organoleptic test and TPA present excellent hardness after Flavourzyme treatment in soy protein that resembles dairy cheese (Table 1, 2 and 4). In addition, slight sticky texture was observed in pea formulation in organoleptic test but adhesiveness by TPA presented low adhesion of the cheese analog. The binding of semi-hard cheese is enhanced compared to native protein in both pea and soy proteins. In addition, the significant change in eliminating sandy texture from cheese formulations after protein incorporation is achieved after enzymatic treatment with Flavourzyme as can be seen from the results.

[0253] Shredding of cheese was applicable and presented clean cut edge, uniform shreds, and no clumping.

[0254] The modified Schreiber test presented enhanced significancy the DI% increase after Flavourzyme treatment in pea protein compared to native and similar to dairy cheese, while in soy protein the melting was slightly enhanced in this test and is similar to dairy cheese (Fig 7 and Fig. 8).

[0255] The melting cheese on pizza application presented in Fig. 9 for pea and Fig. 10 for pea and soy cheese shows that native protein formulation did not present good melting for pizza application, while the Flavourzyme treated proteins presented good melting.

[0256] The stretchability in sandwich toast presented an increase after Flavourzyme treatment with pea and soy proteins compared to native protein (Fig 11 - Fig.14).EXAMPLE 7Preparation and characterization of the food product

[0257] Treatment of 10% w / w plant protein in water PPI or SPI with protein glutaminase (PG) or protein deamidase with endo protease. The treatment is similar to example 1. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (15% wt. of the total wt.) with modified starch (14% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (53.9% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.8% wt. of the total wt.) were mixed in a speed of 2 until homogenized.The cheese analog preparation and storage are similar to example 1.EXAMPLE 8Preparation and characterization of the food product

[0258] Treatment of 10% w / w plant protein in water PPI or SPI with protein glutaminase (PG) or protein deamidase with endo protease. The treatment is similar to example 1. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (10% wt. of the total wt.) with acetylated potato starch (17% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (50.9% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.8% wt. of the total wt.) were mixed in a speed of 2 until homogenized.The cheese analog preparation and storage are similar to example 1.EXAMPLE 9Preparation and characterization of the food product

[0259] Treatment of 10% w / w plant protein in water PPI or SPI with protein glutaminase (PG) or protein deamidase with endo protease. The treatment is similar to example 1. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (10% wt. of the total wt.) with native waxy maize starch (17% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (50.9% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.8% wt. of the total wt.) were mixed in a speed of 2 until homogenized.The cheese analog preparation and storage are similar to example 1.EXAMPLE 10Preparation and characterization of the food product

[0260] Treatment of 10% w / w plant protein in water PPI or SPI with protein glutaminase (PG) or protein deamidase with endo protease. The treatment is similar to example 1. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (10% wt. of the total wt.) with mixture of acetylated potato and waxy maize starch (17% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (50.9% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.8% wt. of the total wt.) were mixed in a speed of 2 until homogenized.The cheese analog preparation and storage are similar to example 1.EXAMPLE 11Preparation and characterization of the food product

[0261] Treatment of 10% w / w plant protein in water PPI or SPI with protein glutaminase (PG) or protein deamidase with endo protease. The treatment is similar to example 1. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (10% wt. of the total wt.) with enzymaticallytreated potato starch (17% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (50.9% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.8% wt. of the total wt.) were mixed in a speed of 2 until homogenized.The cheese analog preparation and storage are similar to example 1.EXAMPLE 12Preparation and characterization of the food product

[0262] Treatment of 10% w / w plant protein in water PPI or SPI with protein glutaminase (PG) or protein deamidase with endo protease. The treatment is similar to example 1. The modified protein solution was freeze dried or spray dried to obtain modified protein hydrolysate powder which was then mixed (10% wt. of the total wt.) with acetylated potato starch (14% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.). In a thermomix, water (51.5% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), and flavor (0.2% wt. of the total wt.) were mixed in a speed of 2 until homogenized.Then, the dry mixture of modified protein, starch, LBG and salt was added to the thermomix, mixed at a speed of 4 for few minutes or until homogenized. Finally, melted oil (coconut and sunflower oil mixture, 23% wt. of the total wt.) was poured into the mixture gradually while mixing at speed 4 until homogenized. The mixture was heated to 85 °C for 5 minutes at speed 4 then poured hot into a mold and let it cool to RT then placed in refrigerator 4 to 8 °C for 7-10 days to obtain a shaped product.EXAMPLE 13Preparation and characterization of the food product

[0263] Treatment of 10% w / w fava bean concentrate, pea protein concentrate, or soy protein concentrate in distilled water containing 500 ppm sodium benzoate was pretreated with PG or protein deamidase with endo protease at a temperature between 40 and 60°C. The treatment and cheese analog preparation are similar to Example 1.EXAMPLE 14Preparation and characterization of the food product

[0264] Treatment of 10% w / w fava bean concentrate, pea protein concentrate, or soy protein concentrate in distilled water containing 500 ppm sodium benzoate was pretreated with papain at a temperature between 40 and 60°C. The treatment and cheese analog preparation are similar to Example 1.EXAMPLE 15Preparation and characterization of the food product

[0265] Treatment of 10% w / w fava bean concentrate, pea protein concentrate, or soy protein concentrate in distilled water containing 500 ppm sodium benzoate was pretreated with Endo-SternEnzyme Bp at a temperature between 40 and 60°C. The treatment and cheese analog preparation are similar to Example 1.EXAMPLE 16Preparation and characterization of the food product

[0266] Treatment of 10% w / w fava bean concentrate, pea protein concentrate, or soy protein concentrate in distilled water containing 500 ppm sodium benzoate was pretreated with Protease Neutral P at a temperature between 40 and 60°C. The treatment and cheese analog preparation are similar to Example 1.EXAMPLE 17Preparation and characterization of the food product

[0267] Treatment of 10% w / w fava bean concentrate, pea protein concentrate, or soy protein concentrate in distilled water containing 500 ppm sodium benzoate was pretreated with Flavourzyme at a temperature between 40 and 60°C. The treatment and cheese analog preparation are similar to Example 1.EXAMPLE 18Preparation and characterization of the food product

[0268] Treatment of commercially available tofu (13% soy protein, 7% fat, 4% carbs and fibers and 76% water) with PG or protein deamidase with endo protease activity at a temperature between 40 and 60°C. For this, water was added to grounded tofu for a final soy protein concentration of 10%, the enzymes were added and allowed to mix at a temperature between 40 and 60°C. At the end of the reaction the PG or protein deamidase with endo protease were inactivated by heat by immersing the reaction mixture container in a water bath and reach 90°C for 15min.

[0269] The product was then mixed (69.5% wt. of the total wt.) with modified starch (17.9% wt. of the total wt.), LBG (0.2% wt. of the total wt.) and salt (0.8% wt. of the total wt.), potassium sorbate (food preservative 0.1% wt. of total wt.), lactic acid (0.2% wt. of the total wt.), flavor (0.8% wt. of the total wt.), and melted oil (coconut oil, 10.5% wt. of the total wt.) were mixed in a speed of 4 until homogenized.

[0270] The mixture was heated to 85 °C for 5 minutes at speed 4 then poured hot into a mold and let it cool to RT then placed in refrigerator 4 to 8 °C for 7-10 days to obtain a shaped product.Table 1: Organoleptic properties of semi-hard cheese prepared with PPI and SPI.* Score 10 is high and 0 is low at each property.Table 2: TPA results of PPI cheese with PG and papain in different DH%.Table 3: TPA results of PPI cheese with different enzyme treatments.Table 4: TPA results of SPI cheese with PG and papain in different DH%.Table 5: TPA results of SPI cheese with different enzyme treatments.Table 6: pH of semi-hard cheese prepared with PPI and SPITable 7. Diameter increase in modified Schreiber test.Table 8. Diameter increase in modified Schreiber test of PPI after enzymatic treatment.Table 9. Melting cheese on pizza of PPI and SPI visually scored.*Score 1 in good melting, 0 bad or no melting.

Claims

CLAIMSWhat is claimed is:

1. A food product comprising (i) a modified protein, (ii) a polysaccharide and (iii) a fat; wherein: said (i) and said (ii) are non-crosslinked; the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 5 and 40% or between 0.5 and 30%; and optionally is further characterized by a molar ratio between glutamine and glutamic acid residues of below 1 : 10; said food product is a homogenous solid at a temperature below a melting onset point, and said melting onset point is at a temperature ranging between 30 and 40°C.

2. The food product of claim 1, wherein in a hydrated state said food product comprises said modified protein at a concentration between 2 and 60%w / w, and said fat at a concentration between 5 and 50%'W / w, relative to the total weight of said food product.

3. The food product of claim 1 or 2, wherein a weight ratio between the modified protein and said polysaccharide is between 1: 10 and 10:1, and wherein the homogenous solid is substantially devoid of phase separation.

4. The food product of any one of claims 1 to 3, wherein said fat is or comprises a an oil, saturated fatty acid, unsaturated fatty acid, fatty acid mono-, di- and / or triglyceride, fatty acid phosphate ester, and phospholipid, or any combination thereof.

5. The food product of any one of claims 1 to 3, wherein said polysaccharide is or comprises a modified starch, a native starch, or any combination thereof.

6. The food product of any one of claims 1 to 4, wherein the modified protein is derived from a plant protein; wherein the hydrolysis degree of the modified protein is between about 5 and about 20% or between about 1 and about 25%; and wherein the food product is a dairy food product substitute.

7. The food product of claim 6, wherein the dairy food product substitute is a cheese substitute.

8. The food product of any one of claims 1 to 6, characterized by at least one of: a hardness between 5 and 40N, an adhesiveness between 0.1 and 3Nmm, and a fracture between 5 and 30N, determined by Texture Profile Analyzer.. The food product of any one of claims 5 to 7, wherein said derived is by exposing the plant protein to a first enzyme having a deamidase activity and optionally to a second enzyme having a proteolytic activity.

10. The food product of any one of claims 1 to 8, wherein the modified protein is characterized by at least one of: (i) alpha helix content of at least 50%; (ii) average MW below 120kDa; (iii) water solubility above 25%, as determined based on total nitrogen content; and wherein a weight ratio between the modified protein and said polysaccharide is between 1: 1 and 1:5.

11. The food product of any of claims 1 to 10, wherein said food product in a hydrated state is characterized by a water content between 30 and 70% w / w, and wherein a weight content of the modified protein by dry weight of the food product is between about 10 and about 20%.

12. The food product of any of claims 1 to 11, said food product is characterized by meltability, wherein said meltability comprises a diameter increase between 170 and 350%, as determined by Schreiber test; wherein said polysaccharide is starch; optionally wherein a weight ratio between the modified protein and said polysaccharide is between 1: 1 and 1:3.

13. A method for manufacturing said food product of any one of claims 1 to 12, comprising:(i) contacting an aqueous composition comprising a protein with (a) an enzyme having a deamidase and a proteolytic activity; or (b) a first enzyme having a deamidase activity and with a second enzyme having a proteolytic activity, thereby obtaining the modified protein;(ii) mixing the modified protein or the composition comprising thereof with the polysaccharide and fat, thereby obtaining said food product.

14. The method of claim 13, wherein said step (i) is performed at a temperature between 10 and 70°C; and wherein said step (ii) comprises mixing at a temperature between 40 and 110 °C for a period of time sufficient for forming said food product.

15. The method of any of claims 13 to 14, wherein step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:2 and 10: 1;optionally step (ii) farther comprises adding one or more additional agents selected from flavoring agent, pigment, hydrocolloid, coloring agent, preservative, and salt.

16. The method of any one of claims 13 to 15, wherein the aqueous composition is substantially devoid of the polysaccharide.

17. The method of any one of claims 13 to 16, wherein the modified protein is characterized by a hydrolysis degree between 5 and 40% or between 0.5 and 30%; and by a molar ratio between glutamine and glutamic acid residues of below 1: 10; and by at least one of: alpha helix content of at least 50%; average MW below 120kDa; water solubility above 25%, as determined based on total nitrogen content.

18. The method of any one of claims 13 to 17, wherein a concentration of the protein and / or of the modified protein within the aqueous composition is between 3 and 30%w / w; and wherein the modified protein is characterized by a hydrolysis degree between 1 and 25%.

19. The method of claim 18, wherein at a concentration of the protein within the aqueous composition of about 20%w / w said aqueous composition is a non-flowable composition, and wherein the composition obtained after completion of step (i) is in a form of a flowable liquid.

20. The method of any one of claims 13 to 19, wherein said step (i) comprises adding to the aqueous composition 0.01%-3%w / w of the first enzyme relative to the protein; optionally wherein the first enzyme comprises amidohydrolase, deamidase, protein glutaminase, or any combination thereof.

21. The method of any of claims 13 to 20, further comprising performing a termination step thereby substantially arresting enzymatic activity of said a or of said b, wherein said termination step is performed prior to said step (ii).

22. The method of claim 21, wherein said termination step comprises any of: heating the aqueous composition to a temperature of at least 90°C; adding to the aqueous composition an inhibitor of said a or of said b; or exposing the aqueous composition to an acid to obtain a pH of below 4 or above 9.

23. A method for manufacturing said food product of any one of claims 1 to 12, comprising:(i) contacting an aqueous composition comprising a protein with an enzyme having a proteolytic activity and optionally an additional enzyme having deamidase activity, thereby obtaining an aqueous composition comprising the modified protein;(ii) mixing the modified protein or the aqueous composition with the polysaccharide and fat, thereby obtaining said food product; and wherein the aqueous composition is substantially devoid of a polysaccharide.

24. The method of claim 23, wherein the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 3 and 40% or between 0.5 and 30%.

25. The method of claim 23 or 24, wherein the modified protein is further characterized by at least one of: alpha helix content of at least 50%; average MW below 120kDa; hydrolysis degree between 1 and 20%; and water solubility above 25%, as determined based on total nitrogen content.

26. The method of any one of claims 23 to 25, wherein a concentration of the protein within the aqueous composition is between 3 and 30%w / w, and wherein the aqueous composition consists essentially of said protein.

27. The method of any one of claims 23 to 26, wherein said step (i) comprises adding to the aqueous composition between 0.01%-3%w / w of the enzyme relative to the protein.

28. The method of any one of claims 23 to 27, wherein step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:2 and 10:1.

29. The method of any one of claims 23 to 27, wherein step (ii) comprises a weight ratio between the modified protein and the polysaccharide between 1:3 and 3:1.

30. The method of any one of claims 23 to 29, wherein step (i) further comprises mixing at a temperature between 10 and 70°C.

31. The method of any one of claims 13 to 30, wherein step (i) further comprises performing a termination step, wherein said termination step is performed prior to said step (ii).

32. The method of claim 31, wherein said termination step comprises any of: providing said aqueous composition to a temperature of at least 90°C; adding an inhibitor ofsaid enzyme to said aqueous composition; and exposing said aqueous composition to a pH below 4 or above 9.

33. The method of any of claims 13 to 32, wherein the step (i) further comprises isolating the modified protein from said aqueous composition.

34. The method of claim 33, wherein said isolating is by drying said aqueous composition to obtain a powder comprising the modified protein.

35. The method of any of claims 13 to 34, wherein said step (ii) further comprises drying said food product, optionally wherein said drying is by lyophilization or spray drying.

36. The method of any of claims 13 to 35, wherein said polysaccharide is or comprises starch.

37. The method of any of claims 23 to 36, wherein the enzyme having a proteolytic activity comprises or is selected from any one of: pepsin, trypsin, chymotrypsin, papain, endoprotease, exoprotease, carboxypeptidase, aminopeptidase, protein deamidase or any combination thereof.

38. The method of any one of claims 13 to 36, wherein said protein is or comprises any one of: a plant protein, a plant protein isolate, a concentrated wet plant protein and a precipitated wet plant protein.

39. The method of any one of claims 13 to 38, wherein the modified protein has at least 15% greater water solubility compared to said protein, as determined based on total nitrogen content; and wherein the modified protein has a hydrolysis degree between about 2 and about 25%.

40. The method of any one of claims 36 to 39, wherein said starch is or comprises a modified and / or non-modified starch, optionally wherein the starch comprises or is selected from any of: potato starch, corn starch, rice starch and tapioca starch, or any combination thereof; and wherein a weight ratio between the modified protein and the polysaccharide is between 1:1 and 1:5.

41. A modified protein derived from a plant protein, wherein the modified protein is a hydrolyzed protein characterized by a hydrolysis degree between 0.5 and 30% and by a molar ratio between glutamine and glutamic acid residues below 1: 10; wherein the modified protein has a water solubility above 25%w / w, as determined based ontotal nitrogen content and is characterized by at least one of: (i) alpha helix content of at least 50%; and (ii) average MW below 120kDa.

42. The modified protein of claim 41, wherein said derived is by exposing the plant protein to a first enzyme having a deamidase activity and optionally to a second enzyme having a proteolytic activity.

43. The modified protein of any one of claims 41 to 42, wherein the modified protein is characterized by a hydrolysis degree between about 1.5 and about 25%; molar ratio between glutamine and glutamic acid residues between 1:10 and 1 : 1000; alpha helix content of at least 50% ; and by average MW below 80kDa.

44. A food product comprising the modified protein of any one of claims 41 to 43, (ii) a polysaccharide and (iii) a fat; wherein: said (i) and said (ii) are non-crosslinked; said food product is a homogenous solid at a temperature below a melting onset point, and said melting onset point is at a temperature ranging between 30 and 40°C.

45. The food product of claim 44, wherein in a hydrated state said food product comprises said modified protein at a concentration between 2 and 60%w / w, and said fat at a concentration between 5 and 50%w / w; wherein a weight ratio between the modified protein and said polysaccharide is between 1: 1 and 1: 10, and wherein the homogenous solid is substantially devoid of phase separation.

46. The food product of claim 44 or 45, wherein the food product is a cheese substitute and is characterized by at least one of: a hardness between 5 and 40N, a adhesiveness between 0.1 and 3Nmm, and a fracture between 5 and 30N, determined by Texture Profile Analyzer; and wherein the cheese substitute has a meltability comprising a diameter increase between 170 and 350%, as determiner! by Schreiber test.

47. The food product of claim 7 or 46, wherein the cheese substitute is selected from semi-hard and hard cheese substitute.

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