WO2023166491A1

WO2023166491A1 - Alkaline functionalization of plant-based protein compositions

Info

Publication number: WO2023166491A1
Application number: PCT/IB2023/052018
Authority: WO
Inventors: Jochen Weiss; Hanna Salminen; Laura SCHEUER
Original assignee: Hohenheim, University of
Current assignee: Hohenheim, University of
Priority date: 2022-03-03
Filing date: 2023-03-03
Publication date: 2023-09-07
Anticipated expiration: 2024-09-03

Abstract

The present disclosure provides compositions suitable for use in food products wherein the compositions provide textures to the food products comparable to animal-based food products. Embodiments of the disclosure herein provide for compositions having a plant-based fiber structure capable of undergoing structural changes under alkaline conditions.

Description

ALKALINE FUNCTIONALIZATION OF PLANT-BASED PROTEIN COMPOSITIONS AND METHODS OF USE THEREOF

CROSS REFERENCE TO RELATED APPLICATIONS

[001] This application claims the benefit of the U.S. Provisional application number 63/316,197 filed March 3, 2022 the disclosures of which is herein incorporated by reference in its entirety.

FIELD

[002] Compositions suitable for use in food products comprising one or more plant protein isolates having fiber structures analogous to animal proteins, methods of making plant protein fiber networks, and methods of use thereof, wherein the compositions provide textures to the food products comparable to animal-based food products.

BACKGROUND

[003] Consumer demand for alternatives to animal-based foods, such as meat, eggs, and milk, continues to rise because of growing environmental, health, and ethical concerns associated with the rearing and slaughter of livestock animals. In response to these demands, the food and biotechnology sectors have developed a number of innovative approaches to designing animal- free food products; however, many of these non-animal based alternatives to animal proteins do not fully emulate the sensory and functional properties of animal proteins.

[004] Current difficulties with creating non-animal sourced food products include generating products having the appropriate textures and mouth feel. The structure of proteins plays an essential role in plant-based food products, particularly those that mimic an animal-based food product like dairy or meat. Vegetable proteins usually do not form network or fiber structures, and are relatively unreactive, thus matrix formation for the production of solid foods with animal protein-like textures remains problematic. Plant-based food products now rely on formation of protein doughs which are brought to a high temperature and conveyed under high pressure through a nozzle (i.e., extrusion). After extrusion, the proteins are denatured and inert, i.e., they only act as fillers. The process is irreversible. Accordingly, there is a need to identify plant protein isolates capable of forming reversible structures analogous to the animal proteins found in animal-based foods such as muscle-based meat products and dairy products. There is a desire to form plant protein products that form networks or fiber structures analogous to animal proteins.

SUMMARY OF THE INVENTION

[005] The present disclosure is based, at least in part, on the discovery that plant proteins, in contrast to animal proteins, undergo reversable structural changes under alkaline conditions. The alkaline environment causes the multimeric structure of plant proteins to solubilize into functionalized, reactive proteins capable of reforming into network of fiber structures ~ similar to that of animal proteins. Accordingly, the present disclosure provides novel compositions suitable for use in food products wherein the compositions herein have textures comparable to an animalbased food product.

[006] In certain embodiments, compositions provided herein may comprise at least one plant protein isolate. In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may be stored in an alkaline buffer solution. Accordingly, compositions disclosed herein may form reactive plant proteins when stored in an alkaline buffer solution. Reactive plant proteins may form fiber structures and/or networks analogous to animal proteins in the alkaline buffer solution and/or after the alkaline buffer solution is neutralized. An alkaline buffer solution may be neutralized by reducing the pH of the buffer solution, adding one or more additional proteins, or a combination thereof. Reducing the pH of the buffer solution may be performed in one or more series of steps, wherein each step comprises titrating the buffer solution to a pH lower than the pH resulting from the step proceeding it. In some embodiments, an additional protein added to the reactive plant proteins may be an animal protein, a plant protein, or a combination thereof. The additional protein added to the reactive plant proteins may be a hydrolysate of an animal protein, of a plant protein, or a combination thereof.

[007] In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the total concentration of the at least one plant protein isolate may be at least about 1% w/w. In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the total concentration of the at least one plant protein isolate may be about 1% w/w to about 20% w/w.

[008] In some embodiments, compositions herein may comprise an alkaline buffer solution having a pH equal to or higher than about 11. In some embodiments, compositions herein may comprise an alkaline buffer solution that can comprise water. In some embodiments, compositions herein may comprise an alkaline buffer solution that can comprise at least about 1 % w/w water. In some embodiments, compositions herein may comprise an alkaline buffer solution that can comprise about 1% w/w to about 99% w/w water.

[009] In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may be stored in an alkaline buffer solution for at least about 1 hour. In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may be stored in an alkaline buffer solution for at least about 1 hour to at least about 48 hours.

[010] In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may be stored in an alkaline buffer solution at a temperature of at least about 4 °C. In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may be stored in an alkaline buffer solution at a temperature of about 4 °C to about 140 °C. In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may be stored in an alkaline buffer solution at a temperature of about 20 °C to about 30 °C.

[Oil] In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may comprise a natural protein isolate, a recombinant protein isolate, or any combination thereof. In some embodiments, compositions herein may comprise at least one plant protein isolate wherein the at least one plant protein isolate may comprise one or more proteins present in a crude plant material. In some embodiments, compositions herein may comprise one or more proteins present in a crude plant material wherein the crude plant material can comprise vegetables, fruits, seeds, legumes, grains, or any combination thereof. In some embodiments, compositions herein may comprise one or more proteins present in a crude plant material wherein the crude plant material may comprise pulses. In some embodiments, compositions herein may comprise one or more proteins present in a crude plant material wherein the crude plant material may comprise legumes. In some embodiments, compositions herein may comprise one or more proteins present in a crude plant material wherein the crude plant material may comprise chickpeas, green peas, yellow peas, black eyed peas, pinto beans, kidney beans, black beans, mung beans, soybeans, adzuki beans, fava beans, edamame, green lentils, red lentils, black lentils, lupins, peanuts, or any combination thereof. In some embodiments, compositions herein may comprise one or more proteins present in a crude plant material wherein the crude plant material may comprise peas. In some embodiments, compositions herein may comprise one or more proteins present in a crude plant material wherein the crude plant material may comprise yellow peas.

[012] In some embodiments, compositions herein may comprise at least one plant protein isolate stored in an alkaline buffer solution wherein the at least one plant protein isolate can comprise at least about 10% reactive protein. In some embodiments, compositions herein may comprise at least one plant protein isolate stored in an alkaline buffer solution wherein the at least one plant protein isolate can comprise about 10% to about 100% reactive protein.

[013] In certain embodiments, the present disclosure provides compositions comprised of a plantbased fiber structure. In some embodiments, compositions comprising a plant-based fiber structure herein may comprise at least one plant protein isolate stored at a pH equal to or higher than about 11. In some embodiments, compositions comprising a plant-based fiber structure herein may comprise at least one pea protein isolate stored at a pH equal to or higher than about 11. In some embodiments, compositions comprising a plant-based fiber structure herein may comprise at least one pea protein isolate stored at a pH equal to or higher than about 11 for about 12 hours to about 48 hours.

[014] In some embodiments, compositions comprising a plant-based fiber structure herein may be in a solid structure. In some embodiments, compositions comprising a plant-based fiber structure herein may be a gel. In some embodiments, compositions comprising a plant-based fiber structure herein may be in a solid structure that resembles an animal-based food product. In accordance with these embodiments, an animal-based food product comprising a plant-based fiber structure herein may resemble an animal-based meat product, an animal-based dairy product, or any combination thereof.

[015] In some embodiments, plant-based fiber structures herein may comprise at least about 10% w/w moisture content. In some embodiments, plant-based fiber structures herein may comprise about 10% w/w to about 75% w/w moisture content.

[016] In some embodiments, plant-based fiber structures herein may comprise at least one fiber having a diameter of at least about 1.5 mm. In some embodiments, plant-based fiber structures herein may comprise at least one fiber having a diameter of about 1.5 mm to about 8 mm. In some embodiments, plant-based fiber structures herein may comprise at least one fiber having a length of at least about 2 mm. In some embodiments, plant-based fiber structures herein may comprise at least one fiber having a length of about 2 mm to about 40 mm.

[017] In some embodiments, compositions herein having a plant-based fiber structure may comprise at least one property of animal-based food product. In some embodiments, compositions herein having a plant-based fiber structure may comprise at least one property of animal-based food product selected from a group consisting of an animal-based meat product, an animal-based dairy product, or any combination thereof. In some embodiments, plant-based fiber structures herein may be in a solid structure with a water dispersion equal to that of an animal-based food product. In some embodiments, plant-based fiber structures herein may be in a solid structure with a tensile strength equal to that of an animal-based food product.

[018] In some embodiments, plant-based fiber structures herein may comprise an anisotropic structure. In some embodiments, compositions comprising a plant-based fiber structure herein may change form following alkalinization of the one or more plant proteins that comprises the plantbased fiber structure. In accordance with these embodiments, alkalinization of the one or more plant proteins that comprise the plant-based fiber structure herein may occur at a pH ranging from about 9 to about 12. In accordance with these embodiments, alkalinization of the one or more plant proteins that comprise the plant-based fiber structure herein may occur at a pH equal to or higher than about 11. In some embodiments, compositions comprising a plant-based fiber structure herein may change form after neutralizing the pH of the composition. In accordance with these embodiments, the pH of the composition may be neutralized at a pH ranging from about 6 to about 8.

[019] In some embodiments, compositions comprising a plant-based fiber structure herein may comprise at least one pea protein isolate, wherein the at least one pea protein isolate may comprise a natural pea protein or a recombinant pea protein.

[020] In some embodiments, compositions comprising a plant-based fiber structure herein may further comprise at least one other isolated protein. In some embodiments, compositions comprising a plant-based fiber structure herein may further comprise a casein protein. In some embodiments, compositions comprising a plant-based fiber structure herein may further comprise a casein protein wherein the casein protein may be isolated from an animal source. In some embodiments, compositions comprising a plant-based fiber structure herein may further comprise a casein protein wherein the casein protein may be prepared recombinantly in one or more non- animal sources. In accordance with these embodiments, one or more non-animal sources suitable for preparing recombinant proteins (e.g., casein proteins) may comprise a genetically modified plant, a genetically modified bacteria, a genetically modified yeast, or any combination thereof. In accordance with these embodiments, one or more non-animal sources suitable for preparing recombinant proteins (e.g., casein proteins) may comprise a genetically modified yeast.

[021] The casein protein may comprise one or more casein or casein subunit proteins including one or more of a-casein, as 1 -casein, as2-casein, P-casein, K-casein, para-K-casein or any combination thereof. In various aspects, at least one casein subunit may comprise one or more casein subunits that is free from or substantially free from one of the other subunits, for example comprising a-casein, K-casein (and/or para-K-casein), and/or a combination thereof, free, or substantially free of b-casein; alternatively, the casein subunit(s) may comprise a-casein, P-casein, and/or a combination thereof, free of K-casein or para-K-casein.

[022] In certain embodiments, the present disclosure provides methods of making a plant-based fiber structure. In some embodiments, methods herein may comprise incubating at least one plant protein isolate stored in a buffer solution. In some embodiments, methods herein may comprise incubating at least one pea protein isolate stored in a buffer solution. In some embodiments, methods herein may comprise incubating at least one pea protein isolate stored in a buffer solution for at least 12 hours, wherein the buffer solution has a pH equal to or higher than about 11. In accordance with these embodiments, the least one pea protein isolate may be stored in a buffer solution for about 12 hours to about 48 hours. In some embodiments, at least one pea protein isolate may be stored in a buffer solution having a pH ranging from about 11 to about 14. In some embodiments, methods herein may comprise incubating at least one pea protein isolate stored in a buffer solution at a temperature of about 20 °C to about 30 °C.

[023] In some embodiments, methods of making a plant-based fiber structure may generate a plant-based fiber structure that can mimic at least one property of an animal-based food product comprising an animal-based meat product, an animal-based dairy product, or any combination thereof. In accordance with these embodiments, the least one property of an animal-based food product may comprise color, smell, taste, plasticity, breaking strength, mouth feel, or any combination thereof.

[024] In some embodiments, the current disclosure include a food product comprising: (a) at least one plant protein isolate stored in an alkaline buffer for at least 1 hr prior to incorporation into the food product; (b) a protein isolate selected from a casein protein isolate, a whey protein isolate, or a combination thereof; wherein the pH of the food product is between 6-8. In some embodiments, the food product is a meat replica selected from a meat, poultry or seafood replica. In some embodiments, the food product is a non-dairy milk product selected from a milk, yogurt, ice cream, butter, cheese. In some embodiments, the food product is a liquid or gel composition, selected from a beverage, stew, sauce, paste, spread, or soup. In some embodiments, the at least one plant protein isolate is a pea protein isolate. In some embodiments, the food product further comprises one or more of at an additional protein, a fat, a non-animal-based fat, non-animal- based matrixes, non-animal-based edible fibrous components, emulsifiers, plasticizers, thickeners, surfactants, sugars, coloring agents, binding agents, stabilizing agents, flavor enhancers, flavoring agents, fragrance enhancers, nutritional supplements (e.g. vitamins, minerals, antioxidants), essential oils, pH regulators, preservatives, dietary fibers, gluten, and mixtures thereof.

The current disclosure also encompasses a method of making a food product, the method comprising: obtaining at least one plant protein isolate stored in an alkaline buffer for at least 1 hr; combining the plant protein isolate with a protein isolate selected from casein protein isolate, a whey protein isolate, or a combination thereof. In some embodiments, the food product is a meat replica selected from a meat, poultry or sea food replica. In some embodiments, the food product is a non-dairy milk product selected from a milk, yogurt, ice cream, butter, cheese. In some embodiments, the food product is a liquid or gel composition, selected from a beverage, stew, sauce, paste, spread, or soup. In some embodiments, the method further comprises combining the composition with one or more of an additional protein, a fat, a non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, emulsifiers, plasticizers, thickeners, surfactants, sugars, coloring agents, binding agents, stabilizing agents, flavor enhancers, flavoring agents, fragrance enhancers, nutritional supplements (e.g. vitamins, minerals, antioxidants), essential oils, pH regulators, preservatives, dietary fibers, gluten, and mixtures thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[025] The following drawings form part of the present specification and are included to further demonstrate certain embodiments of the present disclosure, which can be better understood by reference to the drawing in combination with the detailed description of specific embodiments presented herein. Embodiments of the present inventive concept are illustrated by way of example in which like reference numerals indicate similar elements and in which as follows.

[026] Fig. 1 are photographs of the pea protein isolate powder PISANE C9, the pea protein isolate powder PURIS Pea 870, micellar casein powder, a_s-casein, and micellar casein concentrate.

[027] Fig. 2 depicts a representative schematic of sample preparation according to storage times.

[028] Fig. 3 depicts representative phase diagrams of PISANE C9, P-casein reduced micellar casein powder, and a mixture thereof wherein structure formation was assessed by visual appearance, in the first screening phase.

[029] Fig. 4 depicts a representative schematic of sample preparation according to pH and storage times.

[030] Fig. 5A depict representative images of a PISANE C9 solution immediately after pH adjustment to pH 11.

[031] Fig. 5B depict representative images of a PISANE C9 solution after pH adjustment to pH 11 after 24 hours at 25 °C.

[032] Fig. 6A depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder with 0 hr of mixing for both, wherein structure formation was assessed by visual appearance.

[033] Fig. 6B depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with no prior mixing, wherein structure formation was assessed by visual appearance.

[034] Fig. 6C depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with no prior mixing, wherein structure formation was assessed by visual appearance.

[035] Fig. 6D depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 0 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[036] Fig. 6E depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance. [037] Fig. 6F depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[038] Fig. 6G depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 0 hours of mixing and PISANE C9 with 48 hours of mixing, wherein structure formation was assessed by visual appearance.

[039] Fig. 6H depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[040] Fig. 61 depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with 48 hours of mixing, wherein structure formation was assessed by visual appearance.

[041] Fig. 7A depicts representative phase diagrams of a mixture of PISANE C9 and P-casein reduced micellar casein powder with 0 hr of mixing for both, wherein structure formation was assessed by visual appearance.

[042] Fig. 7B depicts representative phase diagrams of a mixture of PISANE C9 and P-casein reduced micellar casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with no prior mixing, wherein structure formation was assessed by visual appearance.

[043] Fig. 7C depicts representative phase diagrams of a mixture of PISANE C9 and P-casein reduced micellar casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with no prior mixing, wherein structure formation was assessed by visual appearance.

[044] Fig. 7D depicts representative phase diagrams of a mixture of PISANE C9 and P-casein reduced micellar casein powder, after addition of casein after 0 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[045] Fig. 7E depicts representative phase diagrams of a mixture of PISANE C9 and P-casein reduced micellar casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[046] Fig. 7F depicts representative phase diagrams of a mixture of PISANE C9 and P-casein reduced micellar casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance. [047] Fig. 7G depicts representative phase diagrams of a mixture of PISANE C9 and P-casein P- casein reduced micellar casein powder, after addition of casein after 0 hours of mixing and PISANE C9 with 48 hours of mixing, wherein structure formation was assessed by visual appearance.

[048] Fig. 7H depicts representative phase diagrams of a mixture of PISANE C9 and P-casein P- casein reduced micellar casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[049] Fig. 71 depicts representative phase diagrams of a mixture of PISANE C9 and a_s-casein P- casein reduced micellar casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with 48 hours of mixing, wherein structure formation was assessed by visual appearance.

[050] Fig. 8A depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein reduced micellar casein powder with 0 hr of mixing for both, wherein structure formation was assessed by visual appearance.

[051] Fig. 8B depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein reduced micellar casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with no prior mixing, wherein structure formation was assessed by visual appearance.

[052] Fig. 8C depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein reduced micellar casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with no prior mixing, wherein structure formation was assessed by visual appearance.

[053] Fig. 8D depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein reduced micellar casein powder, after addition of casein after 0 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[054] Fig. 8E depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein reduced micellar casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[055] Fig. 8F depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein reduced micellar casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance. [056] Fig. 8G depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein P-casein reduced micellar casein powder, after addition of casein after 0 hours of mixing and PISANE C9 with 48 hours of mixing, wherein structure formation was assessed by visual appearance.

[057] Fig. 8H depicts representative phase diagrams of a mixture of PURIS Pea 870 and P-casein P-casein reduced micellar casein powder, after addition of casein after 24 hours of mixing and PISANE C9 with 24 hours of mixing, wherein structure formation was assessed by visual appearance.

[058] Fig. 81 depicts representative phase diagrams of a mixture of PURIS Pea 870 and a_s-casein P-casein reduced micellar casein powder, after addition of casein after 48 hours of mixing and PISANE C9 with 48 hours of mixing, wherein structure formation was assessed by visual appearance.

[059] Fig. 9A depicts representative images of structure formation and phase diagrams of mixtures of PISANE C9 with P-casein micellar casein powder, wherein structure formation was assessed by visual appearance.

[060] Fig. 9B depicts representative images of structure formation and phase diagrams of mixtures of PISANE C9 with a s-casein powder, wherein structure formation was assessed by visual appearance.

[061] Fig. 9C depicts representative images of structure formation and phase diagrams of mixtures of PISANE C9 with P-casein reduced micellar casein concentrate, wherein structure formation was assessed by visual appearance.

[062] Fig. 10A depicts representative images of structure formation and phase diagrams of mixtures of PISANE C9 (10%) wherein structure formation was assessed by visual appearance.

[063] Fig. 10B depicts representative images of structure formation and phase diagrams of mixtures of PURIS Pea 870 (10%), wherein structure formation was assessed by visual appearance.

[064] Fig. 11A depicts representative images of structure formation and phase diagrams of mixtures of PISANE C9 with P-casein micellar casein powder, wherein structure formation was assessed by visual appearance. [065] Fig. 11B depicts representative images of structure formation and phase diagrams of mixtures of PISANE C9 with a_s-casein powder, wherein structure formation was assessed by visual appearance.

[066] Fig. 11C depicts representative images of structure formation and phase diagrams of mixtures of or PURIS Pea 870 with 0-casein micellar casein powder, wherein structure formation was assessed by visual appearance.

[067] Fig. 12 depicts a representative schematic of sample preparation according to pH and storage times after acid hydrolysis of casein at pH 2.

[068] Fig. 13A depicts representative phase diagrams of a mixture of PISANE C9 and casein concentrate after PISANE C9 was stored for 24 h at pH 2, wherein structure formation was assessed by visual appearance.

[069] Fig. 13B depicts representative phase diagrams of a mixture of PISANE C9 and casein concentrate after PISANE C9 was stored for 48 h at pH 2, wherein structure formation was assessed by visual appearance.

[070] Fig. 13C depicts representative phase diagrams of a mixture of PURIS Pea 870 and casein concentrate after PURIS Pea 870 was stored for 24 h at pH 2, wherein structure formation was assessed by visual appearance.

[071] Fig. 13D depicts representative phase diagrams of a mixture of PURIS Pea 870 and casein concentrate after PURIS Pea 870 was stored for 48 h at pH 2, wherein structure formation was assessed by visual appearance.

[072] Fig. 13E depicts representative phase diagrams of a mixture of PISANE C9 and casein powder after PISANE C9 was stored for 24 h at pH 2, wherein structure formation was assessed by visual appearance.

[073] Fig. 13F depicts representative phase diagrams of a mixture of PISANE C9 and casein powder after PISANE C9 was stored for 48 h at pH 2, wherein structure formation was assessed by visual appearance.

[074] Fig. 13G depicts representative phase diagrams of a mixture of PURIS Pea 870 and casein powder after PURIS Pea 870 was stored for 24 h at pH 2, wherein structure formation was assessed by visual appearance. [075] Fig. 13H depicts representative phase diagrams of a mixture of PURIS Pea 870 and casein powder after PURIS Pea 870 was stored for 48 h at pH 2, wherein structure formation was assessed by visual appearance.

[076] Fig. 14A depict representative phase diagrams of samples with previous acidification of casein at pH 2 for 48 hours, of PISANE C9 x micellar casein powder.

[077] Fig. 14B depict representative phase diagrams of samples with previous acidification of PURIS Pea 870 x micellar casein powder.

[078] Fig. 14C depict representative phase diagrams of samples with previous acidification of casein at pH 2 for 48 hours, of PISANE C9 x micellar casein concentrate.

[079] Fig. 14D depict representative phase diagrams of samples with previous acidification of casein at pH 2 for 48 hours, of PURIS Pea 870 x micellar casein concentrate at pH 2-11.

[080] Fig. 15A depict representative phase diagrams of samples with previous homogenization of both protein solutions and acidification of casein at pH 2 of PISANE C9 x micellar casein powder for 24 hours.

[081] Fig. 15B depict representative phase diagrams of samples with previous homogenization of both protein solutions and acidification of casein at pH 2 of PISANE C9 x micellar casein powder for 48 hours.

[082] Fig. 15C depict representative phase diagrams of samples with previous homogenization of both protein solutions and acidification of casein at pH 2 of PISANE C9 x micellar casein concentrate at pH 2-11 for 24 hours.

[083] Fig. 15D depict representative phase diagrams of samples with previous homogenization of both protein solutions and acidification of casein at pH 2 of PISANE C9 x micellar casein concentrate at pH 2-11 for 48 hours.

[084] Fig. 16A depicts representative microscopic images of samples of pea protein (PISANE C9) (10% w/w), a mixture of pea protein (5% w/w) and casein concentrate (5% w/w) and casein concentrate (5% w/w) (total protein content 10% w/w) stored at pH 2-6 for 0 to 24 hrs. The scale bar is 200 pm.

[085] Fig. 16B depicts representative microscopic images of samples of pea protein (PISANE C9) (10% w/w), a mixture of pea protein (5% w/w) and casein concentrate (5% w/w) and casein concentrate (5% w/w) (total protein content 10% w/w) stored at pH 7-11 for 0 to 24 hrs. The scale bar is 200 pm. [086] Fig. 17A depicts representative microscopic images of pea protein (PISANE C9) mixed with cis-casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7- 11.

[087] Fig. 17B depicts representative microscopic images of pea protein (PISANE C9) mixed with as-casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7- 11.

[088] Fig. 17C depicts representative microscopic images of pea protein (PISANE C9) mixed with as-casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7- 11.

[089] Fig. 17D depicts representative microscopic images of pea protein (PISANE C9) mixed with as-casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7- 11.

[090] Fig. 17E depicts representative microscopic images of pea protein (PISANE C9) mixed with as-casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7- 11.

[091] Fig. 17F depicts representative microscopic images of pea protein (PISANE C9) mixed with as-casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7- 11.

[092] Fig. 18A depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[093] Fig. 18B depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[094] Fig. 18C depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[095] Fig. 18D depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11. [096] Fig. 18E depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[097] Fig. 18F depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[098] Fig. 19A depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[099] Fig. 19B depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1 : 1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[0100] Fig. 19C depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1:1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[0101] Fig. 19D depicts representative microscopic images of pea protein (PISANE C9) mixed with p-casein reduced micellar casein powder at a ratio 1:1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[0102] Fig. 19E depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1:1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[0103] Fig. 19F depicts representative microscopic images of pea protein (PISANE C9) mixed with P-casein reduced micellar casein powder at a ratio 1:1 (total protein content 10% w/w) for indicated times at pH ranges 7-11.

[0104] Fig. 20A depicts representative microscopic images of samples of micellar casein powder (5% w/w) mixed with pea protein (PISANE C9) (5% w/w) after being hydrolyzed at pH 2-4 for 24 hours and 48 hours at a ratio 1: 1 (total protein content 10% w/w). The scale bar is 200 pm.

[0105] Fig. 20A depicts representative microscopic images of samples of micellar casein powder (5% w/w) mixed with pea protein (PISANE C9) (5% w/w) after being hydrolyzed at pH 4-8 for 24 hours and 48 hours at a ratio 1: 1 (total protein content 10% w/w). The scale bar is 200 pm. [0106] Fig. 20A depicts representative microscopic images of samples of micellar casein powder (5% w/w) mixed with pea protein (PISANE C9) (5% w/w) after being hydrolyzed at pH 8-11 for 24 hours and 48 hours at a ratio 1: 1 (total protein content 10% w/w). The scale bar is 200 pm.

[0107] Fig. 21A depicts representative microscopic images of samples of micellar casein (5% w/w) mixed with pea protein (PURIS Pea 870) (5% w/w) after being hydrolyzed at pH 2-6 for 24 hours and 48 hours at a ratio 1: 1 (total protein content 10% w/w). The scale bar is 200 pm.

[0108] Fig. 21B depicts representative microscopic images of samples of micellar casein (5% w/w) mixed with pea protein (PURIS Pea 870) (5% w/w) after being hydrolyzed at pH 7-11 for 24 hours and 48 hours at a ratio 1: 1 (total protein content 10% w/w). The scale bar is 200 pm.

[0109] Fig. 22A depicts representative microscopic images of individual protein samples of pea protein isolate PISANE C9, pea protein isolate PURISpea 870, a_s-casein powder, micellar casein powder, and casein concentrate respectively at indicated pH and storage time.

[0110] Fig. 22B depicts representative microscopic images of individual protein samples of pea protein isolate PISANE C9, pea protein isolate PURISpea 870, a_s-casein powder, micellar casein powder, and casein concentrate respectively at indicated pH and storage time.

[0111] Fig. 22C depicts representative microscopic images of individual protein samples of pea protein isolate PISANE C9, pea protein isolate PURISpea 870, a_s-casein powder, micellar casein powder, and casein concentrate respectively at indicated pH and storage time.

DETAILED DESCRIPTION

[0112] Plant proteins are mainly made of globular proteins which form multimers covalently linked together whereas animal (i.e. meat) proteins have a complex hierarchical construction of fibrous protein bundles. While animal proteins form flexible fibrous protein networks, the globular plant proteins are made up of polypeptide chains that fold into a densely packed shape. Because plant proteins cannot endogenously form fibrous protein networks like animal proteins, the plant proteins must be modified to undergo a structural change in order to reveal reactive side chains, thus allowing for novel protein-protein interactions to emerge. As used herein, plant proteins that undergo “a structural change” refer to a plant protein having a protein structure different than its native structure (e.g., globular protein structure). In some embodiments, a plant protein subjected to any of the methods disclosed herein may have a structural change in its primary protein structure, its secondary protein structure, its tertiary protein structure, its quaternary protein structure, or any combination thereof. Modifications to the plant protein refers to methods resulting in changes in the protein structure. Such structural changes expose reactive proteins thereby creating new or improved functional properties that may mimic animal proteins. Such functional properties can include, but are not limited to gelation, solubility, thermal stability, emulsification, foamability, and the like. Examples of modifications to plant proteins may include, but are not limited to, physical, chemical, biological perturbations, or a combination thereof. The present disclosure is based, at least in part, on the discovery that plant proteins undergo a structural change, especially in the alkaline range. As such, the present disclosure provides functionalized plant proteins with one or more improved or new functional properties resulting from chemical modification (i.e., alkalinization) of the plant protein.

[0113] In animal proteins, supramolecular structures are formed in the acidic range (e.g. the casein micelle in milk, or the formation of fiber structure in meat after slightly acidifying and the addition of salt) so that the proteins become reactive. Solid products (sausage or cheese) result after heating. Plant proteins exist as pairs, trimers or hexamers, and due to their different chemical structure, do not dissolve in the acidic range, but instead dissolve at higher pH values, where they would thus produce reactive proteins. The present disclosure demonstrates that plant proteins dissolved during storage in an alkaline buffer are capable of forming network structures before and after lowering the pH (i.e., neutralization). The present disclosure thus provides compositions that form structures from plant extracts which are analogous to structures formed from animal proteins. In some embodiments, functionalized plant proteins disclosed herein may form proteinaceous networks that resemble an animal tissue, such an animal connective tissues and/or an animal muscle. Functionalized plant proteins prepared as disclosed herein may form fibrous proteinaceous networks that mimic the fiber-like structures which contribute to the textural qualities of animalbased foods. Accordingly, the present disclosure also provides compositions suitable for use in plant-based food products having at least one property of an animal-based food product.

[0114] The following detailed description references the accompanying drawings that illustrate various embodiments of the present inventive concept. The drawings and description are intended to describe embodiments and embodiments of the present inventive concept in sufficient detail to enable those skilled in the art to practice the present inventive concept. Other components can be utilized and changes can be made without departing from the scope of the present inventive concept. The following description is, therefore, not to be taken in a limiting sense. The scope of the present inventive concept is defined only by the appended claims, along with the full scope of equivalents to which such claims are entitled.

I. Terminology

[0115] The phraseology and terminology employed herein are for the purpose of description and should not be regarded as limiting. For example, the use of a singular term, such as, “a” is not intended as limiting of the number of items. Also, the use of relational terms such as, but not limited to, “top,” “bottom,” “left,” “right,” “upper,” “lower,” “down,” “up,” and “side,” are used in the description for clarity in specific reference to the figures and are not intended to limit the scope of the present inventive concept or the appended claims.

[0116] Further, as the present inventive concept is susceptible to embodiments of many different forms, it is intended that the present disclosure be considered as an example of the principles of the present inventive concept and not intended to limit the present inventive concept to the specific embodiments shown and described. Any one of the features of the present inventive concept may be used separately or in combination with any other feature. References to the terms “embodiment,” “embodiments,” and/or the like in the description mean that the feature and/or features being referred to are included in, at least, one embodiment of the description. Separate references to the terms “embodiment,” “embodiments,” and/or the like in the description do not necessarily refer to the same embodiment and are also not mutually exclusive unless so stated and/or except as will be readily apparent to those skilled in the art from the description. For example, a feature, structure, process, step, action, or the like described in one embodiment may also be included in other embodiments but is not necessarily included. Thus, the present inventive concept may include a variety of combinations and/or integrations of the embodiments described herein. Additionally, all embodiments of the present disclosure, as described herein, are not essential for its practice. Likewise, other systems, methods, features, and advantages of the present inventive concept will be, or become, apparent to one with skill in the art upon examination of the figures and the description. It is intended that all such additional systems, methods, features, and advantages be included within this description, be within the scope of the present inventive concept, and be encompassed by the claims.

[0117] As used herein, the term “about,” can mean relative to the recited value, e.g., amount, dose, temperature, time, percentage, etc., ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%. [0118] The terms "comprising," "including," “encompassing” and "having" are used interchangeably in this disclosure. The terms "comprising," "including," “encompassing” and "having" mean to include, but not necessarily be limited to the things so described.

[0119] The terms “or” and “and/or,” as used herein, are to be interpreted as inclusive or meaning any one or any combination. Therefore, “A, B or C” or “A, B and/or C” mean any of the following: “A,” “B” or “C”; “A and B”; “A and C”; “B and C”; “A, B and C.” An exception to this definition will occur only when a combination of elements, functions, steps or acts are in some way inherently mutually exclusive.

[0120] As used herein, "w/w" refers to proportions by weight and means the ratio of the weight of one substance in a composition to the total weight of the composition, or the weight of one substance in the composition to the weight of another substance of the composition. By means of example only, a reference to a composition that comprises plant protein totaling 10% w/w of the composition means that 10% of the composition's weight is composed of plant protein (e.g., such a composition having a weight of 100 mg would contain 10 mg of plant protein) and the remainder of the weight of the composition (e.g., 90 mg in this example) is composed of other ingredients.

[0121] The term “nucleic acid” or “polynucleotide” refers to deoxyribonucleic acids (DNA) or ribonucleic acids (RNA) and polymers thereof in either single- or double-stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid and are metabolized in a manner similar to naturally occurring nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated. Specifically, degenerate codon substitutions may be achieved by generating sequences in which the third position of one or more selected (or all) codons is substituted with mixed-base and/or deoxy inosine residues (Batzer et al., Nucleic Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem. 260:2605-2608 (1985); and Rossolini et al., Mol. Cell. Probes 8:91-98 (1994)).

[0122] The terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein's or peptide's sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. A polypeptide includes a natural peptide, a recombinant peptide, or a combination thereof.

[0123] As used herein, “recombinant” refers to a cell, nucleic acid, protein, or vector, which has been modified due to the introduction of an exogenous nucleic acid or the alteration of a native nucleic acid.

[0124] It should also be understood that, unless clearly indicated to the contrary, in any methods claimed herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited.

II. Compositions

[0125] The present disclosure provides compositions suitable for use in food products herein, comprising have one or more plant-based fiber structures. As used herein, the term “plant-based fiber structure” includes a matrix of plant proteins. In the area of plant-based food products, the structure of proteins plays an essential role; however, plant proteins usually do not form network or fiber structures and are relatively unreactive, so that matrix formation to produce solid foods with interesting textures is presently problematic in the field. In some embodiments, plant-based fiber structures herein may comprise one or more plant proteins. Plant-based fiber structures disclosed herein may comprise one or more plant proteins wherein the one or more plant proteins may exist as pairs, trimers, hexamers, or any combination thereof.

[0126] Plant proteins for use in the plant-based fiber structures herein may be sourced from raw plant material. The term “raw plant material” as used herein can refer to crude plant material that can be converted by processing according to the present disclosure into a new and useful product such as protein isolate containing proteins originally present in the crude plant material. Raw plant materials may include material derived from plants. In some embodiments, the raw plant material may be sourced from a non-genetically modified, commoditized, hybridized, or genetically modified plant. Raw plant materials can include, but are not limited to, vegetables, fruits, seeds, legumes, grains, or any combination thereof. Additionally, raw plant materials suitable for use herein may include soybeans, other beans, legumes, lentils, peas, and any combination thereof. Non-limiting examples of legumes may include chickpea, green pea, yellow pea, black eyed peas, pinto bean, kidney bean, black bean, mung bean, soybean, adzuki bean, fava bean, edamame, green lentil, red lentil, black lentil, lupin, peanut, and combinations thereof. Pulses may also be used herein as raw plant materials. Exemplarily pulses include, non-soybean, non-peanut legumes, such as peas, beans, lentils, and chickpeas.

[0127] In some preferred embodiments, raw plant materials can include peas. As used herein, “pea” means the mostly small spherical seed of the pod fruit Pisum sativum. Peas suitable for use herein may be from varieties of Pisum sativum. Non-limiting examples of peas suitable for use herein may be field peas, yellow peas, green peas, and/or wrinkled peas that are grown to produce dry peas that are shelled from the mature pod. In some embodiments, peas suitable for use herein may be yellow peas.

[0128] Other plant proteins for use in the plant-based fiber structures herein may be a plant protein isolated from a plant material. As used herein, the term “isolated” or “isolating” may refer to a process which separates proteins from said protein comprising fraction. In general, any method of preparing a protein isolate from plant material known in the art is suitable for use herein. Nonlimiting examples of preparing protein isolates from plants can include precipitation, flocculation, filtration, chromatography, alkali extraction/isoelectric precipitation (AE-IP), salt extractiondialysis (SE), micellar precipitation (MP), and the like.

[0129] Plant proteins for use in the plant-based fiber structures herein may be from a genetically modified non-animal source. In accordance with certain embodiments herein, a genetically modified non-animal source may be a genetically modified plant, a genetically modified bacteria, a genetically modified yeast, or any combination thereof. In some embodiments, plant proteins herein may be recombinant plant proteins. As used herein “recombinant plant proteins” refers to plant proteins recombinantly produced using polypeptide expression techniques (e.g., heterologous expression techniques using bacterial cells, insect cells, fungal cells such as yeast, plant cells such as tobacco, soybean, or Arabidopsis, or mammalian cells). In some embodiments, recombinant plant proteins herein may be a polypeptide encoded from a polynucleotide, wherein the polynucleotide may have an endogenous (i.e., wild-type) nucleic acid sequence for a plant protein derived from a plant source as described herein. In some other embodiments, recombinant plant proteins disclosed herein may be isolated from proteins used in preparing the recombinant plant proteins. Non-limiting examples of preparing isolated recombinant plant proteins can include precipitation, filtration, chromatography, and the like.

[0130] One of skill in the art will understand that the plant protein isolates herein may predominantly, but not exclusively comprise the plant protein of interest. Residual impurities may be present in such plant protein isolates. Residual impurities may include but are not limited to minerals, sugars, carbohydrates, and the like. A plant protein isolate herein may comprise at least about 70 wt % plant proteins to at least about 99% wt % plant proteins. In some embodiments, a plant protein isolate herein may comprise at least about 70 wt %, at least about 75 wt %, at least about 80 wt %, at least about 85 wt %, at least about 90 wt %, at least about 95 wt %, or at least about 99 wt % plant proteins. In some other embodiments, a plant protein isolate herein may comprise about 90 wt %, about 91 wt %, about 92 wt %, about 93 wt %, about 94 wt %, about 95 wt %, about 96 wt %, about 97 wt %, about 98 wt %, or about 99 wt % plant proteins.

[0131] A plant protein isolate (e.g., a pea protein isolate) disclosed herein may comprise at least about 40% dry weight plant protein to at least about 99% dry weight plant protein. In accordance with these embodiments, a plant protein isolate herein may comprise about 40% dry weight plant protein to about 99% dry weight plant protein, about 60% dry weight plant protein to about 99% dry weight plant protein, or about 60% dry weight plant protein to about 98% dry weight plant protein. In some embodiments, a plant protein isolate herein may comprise at least one plant protein with a molecular weight of less than about 100 Daltons, less than about 75 Daltons, less than about 50 Daltons, less than about 40 Daltons, less than about 30 Daltons, less than about 20 Daltons, or less than about 10 Daltons. In some other embodiments, a plant protein isolate (e.g., a pea protein isolate) herein may comprise at least one plant protein having a PDCAAS of about 0.6 to about 1.0, about 0.7 to about 1.0, about 0.8 to about 1.0, or about 0.9 to about 1.0.

[0132] Protein denaturation and aggregation are often prerequisites for achieving desired properties and performance of the resulting product (e.g., a food product); therefore, the control of structural changes of the protein is important to developing final product quality attributes. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have one or more reactive proteins. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have one or more reactive proteins capable of forming a plant-based fiber structure disclosed herein. In some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have one or more reactive proteins that forms a plant-based fiber structure by forming a chemical bond with another reactive protein. In still some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have one or more reactive proteins that forms a plant-based fiber structure by forming a chemical bond with another reactive protein, wherein the chemical bond is reversible. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have one or more reactive proteins that forms a plant-based fiber structure wherein the plant-based fiber structure formed is reversible. In some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have at least about 5% (e.g., at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 99%) reactive proteins of the total protein. In still some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may have about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 99% reactive proteins of the total protein.

[0133] Reactive proteins comprising a plant protein isolate disclosed herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition. As used herein, an “alkaline condition” can refer to an environment having a pH higher than about 10. In some embodiments, reactive proteins comprising a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition immediately after exposure to the alkaline condition. Plant protein isolates herein may form an isotropic (e.g., yogurtlike) or an anisotropic (e.g., fibrous structures) gel immediately after exposure to the alkaline condition. In some embodiments, reactive proteins comprising a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition after less than about 1 minute, less than about 10 minutes, less than about 30 minutes, less than about 1 hour, less than about 2 hours, less than about 6 hours, less than about 12 hours, less than about 24 hours, less than about 36 hours, or less than about 48 hours.

[0134] Reactive proteins comprising a plant protein isolate herein (e.g., a pea protein isolate) may also form a plant-based fiber structure in response to an alkaline condition at a temperature of about 0 °C to about 160 °C, about 2 °C to about 150 °C, about 4 °C to about 140 °C, about 6 °C to about 130 °C, about 8 °C to about 120 °C, about 10 °C to about 110 °C, about 12 °C to about 100 °C, about 14 °C to about 95 °C, about 16 °C to about 90 °C, about 18 °C to about 85 °C, or about 20 °C to about 80 °C. In some embodiments, reactive proteins comprising a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition at room temperature (i.e., about 25 °C ± 3 °C).

[0135] Additionally, reactive proteins comprising a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition under pressure. In some embodiments, a plant-based fiber structure may be formed in response to an alkaline condition under absolute pressure ranging from about 1 bar to about 3 bar. In some embodiments, a plant-based fiber structure may be formed in response to an alkaline condition under absolute pressure of about 1 bar, about 1.5 bar, about 2 bar, about 2.5 bar, or about 3 bar. In some other embodiments, a plant-based fiber structure may be formed in response to an alkaline condition under absolute pressure of about 2 bar and a temperature of about 121 °C.

[0136] Plant protein isolates herein (e.g., a pea protein isolate) may form a plant-based fiber structure after exposure to a solution having a pH of about 10, a pH of about 11, a pH of about 12, a pH of about 13, or a pH of about 14. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure after exposure to a solution having a pH of about 10.5, a pH of about 11.0, a pH of about 11.5, or a pH of about 12.0. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure immediately after exposure to a solution having a pH of at least about 10. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in less than about 1 minute, less than about 10 minutes, less than about 30 minutes, less than about 1 hour, less than about 2 hours, less than about 6 hours, less than about 12 hours, less than about 24 hours, less than about 36 hours, or less than about 48 hours after exposure to a solution having a pH of at least about 11. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure after exposure to a solution having a pH of at least about 11 at a temperature of about 0 °C to about 100 °C, about 5 °C to about 95 °C, about 10 °C to about 90 °C, about 15 °C to about 85 °C, or about 20 °C to about 80 °C. In some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plantbased fiber structure after exposure to a solution having a pH of at least about 11 at room temperature (i.e., about 25 °C ± 3 °C).

[0137] A plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure forms a solid structure. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure forms a gel. In some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure forms an emulsion.

[0138] Plant protein isolates herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure is viscous. Viscosity can be measured by methods known in the art, such as for example via a viscometer. Accordingly, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure has a viscosity of about 10 centipoise (cps) to about 5,000,000 cps, about 50 cps to about 3,000,000 cps, about 100 cps to about 2,000,000 cps, or about 500 cps to about 1,000,000 cps. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure has a viscosity of about 50,000 cps to about 2,000,000 cps, about 75,000 cps to about 1,000,000 cps, or about 100,000 cps to about 500,000 cps. In some other embodiments, a plant protein isolate herein (e.g., a pea protein isolate) may form a plant-based fiber structure in response to an alkaline condition wherein the plant-based fiber structure has a viscosity of about 50 cps to about 100,000 cps, about 100 cps to about 80,000 cps, or about 150 cps to about 60,000 cps.

[0139] Plant protein isolates herein (e.g., a pea protein isolate) stored in an alkaline condition may increase gelation time to form a plant-based fiber structure. As used herein, “gelation time” is the amount of time it takes for the composition comprising a plant protein isolate herein to transform into a gel. In some embodiments, a plant protein isolate herein (e.g., a pea protein isolate) in an alkaline condition may have a gelation time of less than about 1 minute, less than about 10 minutes, less than about 30 minutes, less than about 1 hour, less than about 2 hours, less than about 6 hours, less than about 12 hours, less than about 24 hours, less than about 36 hours, or less than about 48 hours.

[0140] Plant-based fiber structures formed in an alkaline condition may be stable at a temperature of about 0 °C to about 100 °C, about 5 °C to about 95 °C, about 10 °C to about 90 °C, about 15 °C to about 85 °C, or about 20 °C to about 80 °C. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may be stable at room temperature (i.e., about 25 °C ± 3 °C). In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may be stable at about a temperature of at least 4 °C for about 1 day, about 3 days, about 5 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 2 months, about 4 months, about 5 months, about 6 months, about 1 year, or about 2 years. In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may be stable at room temperature (i.e., about 25 °C ± 3 °C) for about 1 day, about 3 days, about 5 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 2 months, about 4 months, about 5 months, about 6 months, about 1 year, or about 2 years.

[0141] Additionally, a plant-based fiber structure formed in an alkaline condition herein may be stable after the pH of the condition is changed to a pH of about 6 to about 14, about 7 to about 12, or about 8 to about 11. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may be stable after the pH of the condition is changed to a pH of about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, or about 8.5. In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may be stable for about 1 day, about 3 days, about 5 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 2 months, about 4 months, about 5 months, about 6 months, about 1 year, or about 2 years after the pH of the condition is changed to a pH of about 6 to about 10 (e.g., about 6, about 7, about 8, about 9, about 10). In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may be stable for about 1 day, about 3 days, about 5 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 2 months, about 4 months, about 5 months, about 6 months, about 1 year, or about 2 years after the pH of the condition is changed to a pH of about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, or about 8.5.

[0142] Plant-based fiber structures formed in an alkaline condition herein may comprise at least about 5% w/w moisture content (e.g., at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, or at least about 50% w/w moisture content). In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise about 5% to about 99% w/w moisture content, about 10% to about 95% w/w moisture content, or about 15% to about 90% w/w moisture content. In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 99% w/w moisture content. In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise about 20%, about 25%, about 30%, about 35%, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%, or about 75% w/w moisture content.

[0143] Other plant-based fiber structures formed in an alkaline condition herein may be dehydrated. Accordingly, a dehydrated plant-based fiber structure formed in an alkaline condition herein may comprise less than about 25% w/w moisture content (e.g., less than about 25% w/w moisture, less than about 20% w/w moisture, less than about 15% w/w moisture, less than about 10% w/w moisture, less than about 5% w/w moisture, less than about 1% w/w moisture, less than about 0.5% w/w moisture). In some embodiments, a dehydrated plant-based fiber structure formed in an alkaline condition herein may be rehydrated. A rehydrated plant-based fiber structure formed in an alkaline condition disclosed herein may comprise at least about 5% w/w moisture content (e.g., at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, or at least about 50% w/w moisture content).

[0144] Plant-based fiber structures formed in an alkaline condition herein may retain moisture content when the plant-based fiber structure is heated. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may retain moisture content when the plant-based fiber structure is heated to at least about 40 °C (e.g., at least about 40 °C, at least about 50 °C, at least about 60 °C, at least about 70 °C, at least about 80 °C, at least about 100 °C, at least about 110 °C, at least about 120 °C, at least about 140 °C). In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may retain moisture content when the plantbased fiber structure is heated to a temperature ranging from about 10 °C to about 140 °C, about 20 °C to about 130 °C, about 30 °C to about 120 °C, about 40 °C to about 110 °C, about 50 °C to about 100 °C, about 60 °C to about 90 °C, or about 70 °C to about 80 °C. In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may retain moisture content when the plant-based fiber structure is heated to about 40 °C, about 45 °C, about 50 °C, about 55 °C, about 60 °C, about 65 °C, about 70 °C, about 75 °C, about 80 °C, about 85 °C, about 90 °C, about 95 °C, about 100 °C, about 105 °C, about 110 °C, about 115 °C, about 120 °C, about 125 °C, about 130 °C, about 135 °C, or about 140 °C. In some embodiments, a plantbased fiber structure formed in an alkaline condition herein may lose about 1% to about 40% moisture content (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%) when the plant-based fiber structure is heated. In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may lose about 1% to about 40% moisture content (e.g., about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%) when the plant-based fiber structure is heated to at least about 40 °C (e.g., at least about 40 °C, at least about 50 °C, at least about 60 °C, at least about 70 °C, at least about 80 °C, at least about 90 °C, at least about 100 °C, at least about 110 °C, at least about 120 °C, at least about 130 °C, at least about 140 °C).

[0145] Plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure resembling that of an animal muscle. An animal muscle for comparison to the plant-based fiber structures formed herein can include, but are not limited to muscle fibers of a cow, a sheep, a goat, a pig, a horse, a camel, a chicken, a turkey, a duck, and the like.

[0146] Plant-based fiber structures formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a diameter similar to that of an animal muscle. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a diameter of at least about 0.5 mm (e.g., at least about 0.5 mm, at least about 1.0 mm, at least about 1.5 mm, at least about 2.0 mm, at least about 3.0 mm, at least about 4.0 mm, at least about 5.0 mm). In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a diameter ranging from about 0.5 mm to about 10 mm, about 1.0 mm to about 9 mm, or about 1.5 mm to about 8 mm. In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a diameter of about 0.5 mm, about 1.0 mm, about 1.5 mm, about 2.0 mm, about 2.5 mm, about 3.0 mm, about 3.5 mm, about 4.0 mm, about 4.5 mm, about 5.0 mm, about 5.5 mm, about 6.0 mm, about 6.5 mm, about 7.0 mm, about 7.5 mm, about 8.0 mm, about 8.5 mm, about 9.0 mm, about 9.5 mm, or about 10 mm.

[0147] Additionally, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a length similar to that of an animal muscle. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a length of at least about 0.5 mm (e.g., at least about 0.5 mm, at least about 1.0 mm, at least about 1.5 mm, at least about 2.0 mm, at least about 3.0 mm, at least about 4.0 mm, at least about 5.0 mm). In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a length ranging from about 0.5 mm to about 50 mm, about 1.0 mm to about 45 mm, or about 2.0 mm to about 40 mm. In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise a fiber structure having at least one or more fibers having a length of about 0.5 mm, about 1.0 mm, about 2.0 mm, about 3.0 mm, about 4.0 mm, about 5.0 mm, about 6.0 mm, about 7.0 mm, about 8.0 mm, about 9.0 mm, about 10 mm, about 15 mm, about 20 mm, about 25 mm, about 30 mm, about 35 mm, about 40 mm, about 45 mm, or about 50 mm.

[0148] Plant-based fiber structure formed in an alkaline condition herein may comprise solid structure with a water dispersion equal to that of an animal-based food product. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise solid structure having a homogeneous dispersion of water particles throughout the structure. In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise solid structure having a heterogeneous dispersion of water particles throughout the structure. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise solid structure having a dispersion of water particles throughout the structure wherein the water particles may have a diameter averaging between about 1 m to about 20 m, about 2 pm to about 15 pm, or about 3 pm to about 10 pm in size. In still some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise solid structure having a dispersion of water particles throughout the structure wherein the water particles may have a diameter averaging about 1 pm, about 2 pm, about 3 pm, about 4 pm, about 5 pm, about 6 pm , about 7 pm, about 8 pm, about 9 pm, about 10 pm, about 11 pm, about 12 pm, about 13 pm, about 14 pm, about 15 pm, about 16 pm, about 17 pm, about 18 pm, about 19 pm, or about 20 pm in size.

[0149] Plant-based fiber structure formed in an alkaline condition herein may comprise solid structure with a tensile strength equal to that of an animal-based food product. Methods of measuring tensile strength known in the art are suitable for use herein, including use of a texture analyzer. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise solid structure having a tensile strength ranging from about 0.05 kPa to about 10 kPa. In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may comprise solid structure having a tensile strength of about 0.05 kPa, about 0.1 kPa, about 0.5 kPa, about 1 kPa, about 1.5 kPa, about 2 kPa, about 2.5 kPa, about 3 kPa, about 3.5 kPa, about 4 kPa, about 4.5 kPa, about 5 kPa, about 5.5 kPa, about 6 kPa, about 6.5 kPa, about 7 kPa, about 7.5 kPa, about 8 kPa, about 8.5 kPa, about 9 kPa, about 9.5 kPa, or about 10 kPa.

[0150] Plant-based fiber structures formed in an alkaline condition herein may comprise solid structure with a hardness equal to that of an animal-based food product. Methods of measuring hardness known in the art are suitable for use herein, including use of a texture analyzer. In some embodiments, plant-based fiber structures herein may comprise a solid structure having a hardness ranging from about 150 kPa to about 1000 kPa. In some other embodiments, plant-based fiber structures herein may comprise a solid structure having a hardness of about 150 kPa, about 175 kPa, about 200 kPa, about 225 kPa, about 250 kPa, about 275 kPa, about 300 kPa, about 325 kPa, about 350 kPa, about 375 kPa, about 400 kPa, about 425 kPa, about 450 kPa, about 475 kPa, about 500 kPa, about 525 kPa, about 550 kPa, about 575 kPa, about 600 kPa, about 625 kPa, about 650 kPa, about 675 kPa, about 700 kPa, about 725 kPa, about 750 kPa, about 775 kPa, about 800 kPa, about 825 kPa, about 850 kPa, about 875 kPa, about 900 kPa, about 925 kPa, about 950 kPa, about 975 kPa, or about 1000 kPa.

[0151] Plant-based fiber structures formed in an alkaline condition herein may be combined with one or more proteins. In some embodiments, a plant-based fiber structure herein may be first formed in an alkaline condition followed by addition of one or more proteins. In some other embodiments, a plant-based fiber structure herein may be first formed in an alkaline condition followed by addition of one or more proteins wherein the addition of the one or more proteins lowers the pH of the condition. In still some other embodiments, a plant-based fiber structure herein may be first formed in an alkaline condition followed by addition of one or more proteins wherein the addition of the one or more proteins lowers the pH of the condition to a pH of about 6 to about 9. In some embodiments, a plant-based fiber structure herein may be first formed in an alkaline condition followed by addition of one or more proteins wherein the addition of the one or more proteins lowers the pH to a pH of about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, or about 8.5. In some embodiments, the pH may be further adjusted to a range of about 6 to about 8.5 using any food-grade organic or inorganic buffer including but not limited to potassium metaphosphate, potassium phosphate, potassium phosphate dibasic anhydrous, potassium phosphate monobasic, potassium phosphate tribasic, potassium phosphate dibasic heptahydrate, monobasic sodium acetate, sodium citrate anhydrous and dihydrate, ammonium hydroxide, sodium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate, calcium carbonate, borate, glycine, Tris, Bis-Tris propane, bicine, HEPES, TESm MOBS, DIPSO, TAPS, triethanolamine (TEA), pyrophosphate, HEPPSO, tricine, hydrazine, glycylclycine, 2-amino-2- methyl- 1,3 -propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, tris(hydroxymethyl)aminomethane (THAM), and other materials known to one of ordinary skill in the art. In some embodiments, the buffer is an alkaline buffer for example: ammonium hydroxide, sodium hydroxide, sodium carbonate, ammonium carbonate, calcium carbonate, glycine, TAPS, pyrophosphate, tricine, hydrazine, glycylclycine, 2-amino-2-methy 1-1,3 - propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, or tris(hydroxymethyl)aminomethane (THAM).

[0152] Plant-based fiber structures formed in an alkaline condition herein may be combined with one or more proteins wherein the one or more proteins is a plant-based protein, an animal-based protein, or a combination thereof. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may be combined with one or more proteins wherein the one or more proteins is a recombinantly produced plant-based protein, a recombinantly produced animal-based protein, or a combination thereof.

[0153] Plant-based fiber structures formed in an alkaline condition herein may be combined with one or more proteins wherein the one or more proteins is a casein protein isolate, a whey protein isolate, wheat gluten, soy protein concentrate or pea vicilin or pea legumin or a combination thereof. In some embodiments a plant-based fiber structure formed in an alkaline condition herein may be combined with one or more proteins wherein the one or more proteins is a casein protein isolate wherein the casein protein isolate may comprise alpha-casein peptides, beta-casein peptides, kappa-casein peptides, or any combination thereof. Accordingly, a casein protein isolate for use herein may be derived and/or produced from micellar casein, acid casein, hydrolyzed casein, rennet casein or any combination thereof. Micellar casein is ultrafiltered casein extracted from milk without acidification. Acid casein is a dry free flowing high-quality protein food ingredient that has been isolated from skim milk. Hydrolysed casein is a soluble, enzymatic digest of casein. Rennet casein is produced by the controlled precipitation of casein from pure, pasteurized skim milk through the action of rennet. A casein protein isolate for use herein may be a recombinantly produced casein protein.

[0154] Plant-based fiber structures formed in an alkaline condition herein may be combined with one or more proteins in a ratio of about 1 :1. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may be combined with one or more proteins wherein the about of plant-based fiber structure may be about 1% w/w to about 10% w/w (e.g., about 1% w/w, about 2% w/w, about 3% w/w, about 4% w/w, about 5% w/w, about 6% w/w, about 7% w/w, about 8% w/w, about 9% w/w, about 10% w/w) and the total protein amount of the one or more proteins may be about 1 % w/w to about 10% w/w (e.g., about 1% w/w, about 2% w/w, about 3% w/w, about 4% w/w, about 5% w/w, about 6% w/w, about 7% w/w, about 8% w/w, about 9% w/w, about 10% w/w). In some other embodiments, a plant-based fiber structure formed in an alkaline condition herein may be combined with one or more proteins wherein the about of plant-based fiber structure may be about 5% w/w and the total protein amount of the one or more proteins may be about 5% w/w.

[0155] Plant-based fiber structure compositions herein may have at least one plant protein isolate as disclosed herein. In some embodiments, plant-based fiber structure compositions herein may have at least one pea protein isolate as disclosed herein. In some other embodiments, compositions herein may comprise about 5% to about 99% (e.g., about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%) by weight of the composition of one or more pea protein isolate disclosed herein.

[0156] A buffer solution for use herein can be any solution suitable for use in a food product. In some embodiments, plant-based fiber structure compositions herein may have at least one plant protein isolate as disclosed herein in a buffer solution. In some embodiments, plant-based fiber structure compositions herein may have at least one pea protein isolate as disclosed herein in a buffer solution. In some other embodiments, buffer solutions for use herein may comprise water. In some embodiments, buffer solutions for use herein may comprise at least about 1% w/w water (e.g., at least about 1% w/w water, at least about 5% w/w water, at least about 10% w/w water, at least about 20% w/w water, at least about 30% w/w water, at least about 40% w/w water, at least about 50% w/w water). In some other embodiments, buffer solutions herein may comprise about 1% to about 99% w/w water, about 5% to about 95% w/w water, or about 10% to about 90% w/w water. In still some other embodiments, buffer solutions herein may comprise about 1% w/w water, about 5% w/w water, about 10% w/w water, about 20% w/w water, about 30% w/w water, about 40% w/w water, about 50% w/w water, about 60% w/w water, about 70% w/w water, about 80% w/w water, about 90% w/w water, or about 99% w/w water. [0157] Compositions herein may also include a buffer solution having one or more buffering agents wherein “buffering agents” are compounds used to resist change in pH upon dilution or addition of acid or alkali. Buffering agents for use herein can include, by way of example and without limitation, potassium metaphosphate, potassium phosphate, potassium phosphate dibasic anhydrous, potassium phosphate monobasic, potassium phosphate tribasic, potassium phosphate dibasic heptahydrate, monobasic sodium acetate, sodium citrate anhydrous and dihydrate, ammonium hydroxide, sodium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate, calcium carbonate, borate, glycine, Tris, Bis-Tris propane, bicine, HEPES, TESm MOBS, DIPSO, TAPS, triethanolamine (TEA), pyrophosphate, HEPPSO, tricine, hydrazine, glycylclycine, 2-amino-2-methyl-l,3-propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, tris(hydroxymethyl)aminomethane (THAM), and other materials known to one of ordinary skill in the art. In some embodiments, the buffer is an alkaline buffer for example: ammonium hydroxide, sodium hydroxide, sodium carbonate, ammonium carbonate, calcium carbonate, glycine, TAPS, pyrophosphate, tricine, hydrazine, glycylclycine, 2-amino-2-methyl- 1,3-propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, or tris(hydroxymethyl)aminomethane (THAM). In some embodiments, any food-grade organic or inorganic buffer can be used. In some embodiments, compositions disclosed herein may comprise at least 5%, at least 10%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50% total amount of one or more buffering agents by total weight of the composition. In some other embodiments, the amount of one or more buffering agents may depend on the desired pH level of compositions herein. In some embodiments, compositions disclosed herein may have a pH ranging from about 9 to about 14 (e.g., about 9, about 10, about 11, about 12, about 13, about 14). In some other embodiments, buffer solutions herein may have a pH ranging from about 10 to about 12 (e.g., about 10.0, about 10.5, about 11.0, about 11.5, about 12.0). In some embodiments, buffer solutions herein may have a pH ranging from about 5 to about 8 (e.g., about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0). In some other embodiments, buffer solutions herein may have a pH ranging from about 10 to about 12 (e.g., about 10.0, about 10.5, about 11.0, about 11.5, about 12.0) until a plant-based fiber structure is formed, then the buffer solutions herein may be changed to have a pH ranging from about 5 to about 8 (e.g., about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, about 7.5, about 8.0). [0158] The present disclosure also provides methods of making a plant-based fiber structure. Methods herein may comprise incubating at least one plant protein isolate as disclosed herein in a buffer solution disclosed herein. In some embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution for at least 1 minute, at least 30 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 48 hours. In some other embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution for about 1 minute, at least 30 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 48 hours.

[0159] Additionally, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution wherein the buffer solution may have a pH ranging from about 10 to about 14, or about 11 to about 13. In some embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution wherein the buffer solution may have a pH of at least about 10, at least about 11, at least about 12, at least about 13, or at least about 14. In some other embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution wherein the buffer solution may have a pH of about 10, about 10.5, about 11, about 11.5, about 12, about 12.5, about 13, about 13.5, or about 14.

[0160] Methods of making plant-based fiber structures may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution at a temperature of about 0 °C to about 100 °C, about 5 °C to about 95 °C, about 10 °C to about 90 °C, about 15 °C to about 85 °C, or about 20 °C to about 80 °C. In some embodiments, methods herein may comprise incubating at least one plant protein isolate (e.g., pea protein isolate) in a buffer solution at a temperature of about 10 °C, about 15 °C, about 20 °C, about 25 °C, about 30 °C, or about 35 °C. In some other embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution at room temperature (i.e., about 25 °C ± 3 °C).

[0161] Methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution wherein the buffer solution may have a pH ranging from about 10 to about 14 for at least 1 minute, at least 30 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 48 hours. In some embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution wherein the buffer solution may have a pH ranging from about 10 to about 14 for at least 1 minute, at least 30 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 48 hours at a temperature of about 0 °C to about 100 °C, about 5 °C to about 95 °C, about 10 °C to about 90 °C, about 15 °C to about 85 °C, or about 20 °C to about 80 °C.

[0162] Other methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution wherein the buffer solution may have a pH ranging from about 10 to about 14 for at least 1 minute, at least 30 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 48 hours. In some embodiments, methods herein may comprise incubating at least one plant protein isolate herein (e.g., pea protein isolate) in a buffer solution for at least 12 hours, wherein the buffer solution has a pH equal to or higher than about

I I. Plant protein isolates herein (e.g., pea protein isolate) may be stored in a buffer solution for about 12 hours to about 48 hours. In some other embodiments, at least one plant protein isolate herein may be stored in a buffer solution having a pH ranging from about 11 to about 14. In some embodiments, at least one plant protein isolate herein may be stored in a buffer solution and incubated at a temperature of about 10 °C, about 15 °C, about 20 °C, about 25 °C, about 30 °C, or about 35 °C. In some other embodiments, at least one plant protein isolate herein may be stored in a buffer solution and incubated at a temperature of about 20 °C to about 30 °C. In still some other embodiments, methods herein may comprise incubating at least one plant protein isolate herein in a buffer solution wherein the buffer solution may have a pH of about 11 for at least 1 minute, at least 30 minutes, at least 1 hour, at least 2 hours, at least 6 hours, at least 12 hours, or at least 48 hours at a temperature of about 10 °C, about 15 °C, about 20 °C, about 25 °C, about 30 °C, or about 35 °C.

III. Methods of Use

[0163] The present discourse provides for food products and methods of making such food products using the plant-based fiber structure compositions herein. In certain embodiments, compositions herein can be used for producing meat substitute food products (“meat replicas”). As used herein, a “meat replica” refers to a food product having a realistic meat-like appearance without containing an animal-based competent. Compositions herein can be used as a materials in and in methods of making meat replicas, including, but not limited to ground meat replicas (e.g., ground beef, ground chicken, ground turkey, ground lamb, or ground pork), as well as replicas of cuts of meat and fish. [0164] In some embodiments, the current disclosure also encompasses a method of making a food product, the method comprising: a) obtaining or having obtained at least a plant protein isolate stored in an alkaline buffer for at least 1 hr; combining the plant protein isolate with a protein isolate selected from casein protein isolate, a whey protein isolate, or a combination thereof. In some embodiments, the plant protein isolate stored in an alkaline buffer may comprise a pea protein isolate. In some embodiments, the protein isolate is casein. In some embodiments, the casein is a micellar casein, acid casein, hydrolyzed casein, rennet casein or any combination thereof. In some embodiments, the casein in micellar casein. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may be combined with one or more additional protein isolates (for example: casein, or whey protein), in a weight ratio of the plant protein isolate to additional protein isolate (for example, casein or whey protein) ranging from about 1 : 10 to about 10: 1, or about 1 :5 to about 5:1. In some embodiments, a plant-based fiber structure formed in an alkaline condition herein may be combined with one or more additional protein isolates (for example: casein, or whey protein), in a weight ratio of the plant protein isolate to additional protein isolate (for example, casein or whey protein) ranging from about 1 : 1 to about 1 :2, or 1 :2 to about 1:3, 1:3 to about 1 :4, or 1:4 to about 1 :5, or 1: 1 to about 2: 1, or 2: 1 to about 3:1, or 3: 1 to about 4: 1, or 4: 1 to about 5:1 ratio by weight. In some embodiments, the composition as disclosed herein comprises about 5% (w/w) to about 20% (w/w) of each of the at least a plant protein isolate and the protein isolate. In some aspects, the composition comprises about 5% (w/w), or about 6% (w/w), or about 7% (w/w), or about 8% (w/w), or about 9% (w/w), or about 10% (w/w), or about 11% (w/w), or about 12% (w/w), or about 13% (w/w), or about 14% (w/w), or about 15% (w/w), or about 16% (w/w), or about 17% (w/w), or about 18% (w/w), or about 19% (w/w), or about 20% (w/w) of each of the at least a plant protein isolate and the protein isolate.

[0165] In some embodiments, the composition may have a pH ranging from about 5 to about 8.5. In some embodiments, the addition of the protein isolate to the pea protein isolate lowers the pH of the composition to a pH of about 6.0, about 6.5, about 7.0, about 7.5, about 8.0, or about 8.5. In some embodiments, the pH of the composition may be further adjust to a pH ranging from about 5 to about 8.5, or about 5.5 to about 8, or about 6 to about 7.5, or about 6.5 to about 7. As disclosed herein, the pH of the composition can be adjusted using any food-grade organic or inorganic buffer.

[0166] In some aspects, the method of making the food product further comprises subjecting the composition at least one pea protein isolate subjected to an alkaline buffer and a protein isolate to one or more of a fermentation, cooking, pressurized cooking, extrusion, low-shear, or high shear extrusion process. In some aspects, the composition is subjected to an extrusion process. As used in the current disclosure, an “extrusion” refers to a process in which a material is pushed under compressive stresses through a deformation control element such as a die to form a product. The process of extrusion is usually accomplished by using equipment referred to in the art as an extruder. The extruder as used herein may comprise a single screw extruder or a twin-screw extruder, or a combination thereof. It may be a single screw “wet” extruder (with or without the preconditioner), single screw “dry” extruder (with or without the preconditioner), single-screw interrupted flight extruder (with or without a preconditioner), and twin-screw extruder (with or without a preconditioner). The current disclosure encompasses use of extruders with a wide range of configurations and attachments.

[0167] In some embodiments, the composition may be further combined with additional ingredients for example additional proteins for example a heme-protein, or soy protein, or combination thereof, a fat (milk based like butter), a non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, emulsifiers, plasticizers, thickeners, surfactants, sugars, coloring agents, binding agents, stabilizing agents, flavor enhancers, flavoring agents, fragrance enhancers, nutritional supplements (e.g. vitamins, minerals, antioxidants), essential oils, pH regulators, preservatives, dietary fibers, gluten, and mixtures thereof. In some embodiments, the casein may be a recombinant casein.

[0168] In some embodiments, methods of making food products (e.g., meat replicas) may include combining the compositions herein with non-animal-based fat, non-animal-based matrixes, non- animal-based edible fibrous components, or any combination thereof. Methods of making food products (e.g., meat replicas) may also include combining the compositions herein with non- animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, or any combination thereof wherein the addition of the composition herein provides a meat-like appearance to the meat substitute. In some embodiments, methods of making food products (e.g., meat replicas) may include combining the compositions herein with additional proteins, non- animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, or any combination thereof wherein the addition of the composition herein provides a meat-like taste to the meat substitute. In some other embodiments, methods of making food products (e.g., meat replicas) may include combining the compositions herein with additional non-animal-based proteins, non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, or any combination thereof wherein the addition of the composition herein provides a meat-like smell to the meat substitute. In some embodiments, the additional non-animal-based proteins is a heme-protein, soy protein. In some embodiments, the non-animal-based fat may be selected from soybean oil, canola oil, corn oil, sunflower oil, safflower oil, flaxseed oil, almond oil, peanut oil, fish oil, algal oil, palm oil, palm stearin, palm olein, palm kernel oil, fractionated palm kernel oil (including Medium Chain Triglyceride (MCT) oil made from palm kernel oil), high oleic soybean, canola, sunflower or safflower oils, acai oil, almond oil, amaranth oil, apricot seed oil, argan oil, avocado seed oil, babassu oil, ben oil, blackcurrant seed oil, Borneo tallow nut oil, borage seed oil, buffalo gourd oil, carob pod oil, cashew oil, castor oil, coconut oil, fractionated coconut oil (including Medium Chain Triglyceride (MCT) oil made from coconut oil), coriander seed oil, corn oil, cottonseed oil, evening primrose oil, false flax oil, flax seed oil, grapeseed oil, hazelnut oil, hemp seed oil, kapok seed oil, lallemantia oil, linseed oil, macadamia oil, meadowfoam seed oil, mustard seed oil, okra seed oil, olive oil, palm kernel oil, pecan oil, pequi oil, perilla seed oil, pine nut oil, pistachio oil, poppy seed oil, prune kernel oil, pumpkin seed oil, quinoa oil, ramtil oil, rice bran oil, sesame oil, tea oil, thistle oil, walnut oil, wheat germ oil, hydrogenated palm kernel oil, hydrogenated palm stearin, fully hydrogenated soybean, canola or cottonseed oils, high stearic sunflower oil, enzymatically and chemically interesterified oils, butter oil, cocoa butter, and mixtures thereof.

[0169] In some embodiments, the meat replica food product may further comprise one or more optional ingredients, non-limiting examples of such ingredients include emulsifiers, surfactants, plasticizers, thickeners, sugars, coloring agents, binding agents, stabilizing agents, flavor enhancers, flavoring agents, fragrance enhancers, nutritional supplements (e.g. vitamins, minerals, antioxidants), essential oils, pH regulators, preservatives, dietary fibers, and mixtures thereof. Non-limiting examples of emulsifiers include but are not limited to lecithin, or soy lecithin. Nonlimiting examples of surfactant/solvent components include, Polyoxyethylene sorbitan monostearate (Tween 60), sorbitan monooleate (SMO or Span 80), sorbitan monostearate(SMS or Span 60), glyceryl monooleate (GMO), glyceryl monostearate (GMS) glyceryl monopalmitate (GMP), polyglyceryl ester of lauric acid - polyglyceryl polylaurate (PGPL), polyglyceryl ester of stearic acid - polyglyceryl polystearate (PGPS), polyglyceryl ester of oleic acid (PGPO) - polyglyceryl polyoleate (PGPO), and polyglyceryl ester of ricinoleic acid (PGPR) - polyglyceryl polyricinoleate (PGPR). Non-limiting examples of plasticizers include glycerin or propylene glycol or a combination thereof. Non-limiting examples of thickeners include but not limited to guar gum, pectin, xanthan gum, agar, alginic acid and its salts, carboxymethyl cellulose, carrageenan and its salts, gums, modified starches, pectins, processed Eucheuma seaweed, sodium carboxymethyl cellulose, tara gum. Non-limiting examples of plasticizers include but are not restricted to polysaccharides and galactomannans such as starch, modified starch, maltodextrin, carrageenan, guar gum, alginin, agar, grain flour mix, carboxymethyl cellulose, pectin, locust beam gum and xanthan gum. Non-limiting examples of sugars/sweeteners include but are not limited to stevia, sucralose, sugar alcohols, sucrose, glucose, fructose, and aspartame. Non-limiting examples of a suitable colorant include FD&C colors, such as blue no. 1, blue no. 2, green no. 3, red no. 3, red no. 40, yellow no. 5, yellow no. 6, and the like; natural colors, such as roasted malt flour, caramel coloring, annatto, chlorophyllin, cochineal, betanin, turmeric, saffron, paprika, lycopene, elderberry juice, pandan, butterfly pea and the like, titanium dioxide, and any suitable food colorant known to the skilled artisan. Non-limiting examples of binding agents include but are not restricted to Examples of suitable binding agents include but are not limited to purees (e.g., bean puree, sweet potato puree, pumpkin puree, applesauce, yam puree, banana puree, plantain puree, date puree, prune puree, fig puree, zucchini puree, carrot puree, coconut puree), native or modified starches (e.g., starches from grains, starches from tuber, potato starch, sweet potato starch, corn starch, waxy corn starch, tapioca starch, tapioca, arrowroot starch, taro starch, pea starch, chickpea starch, rice starch, waxy rice starch, lentil starch, barley starch, sorghum starch, wheat starch, and physical or chemical modifications thereof (including, e.g., pre-gelatinized starch, acetylated starch, phosphate bonded starch, carboxymethylated starch, hydroxypropylated starch), flours derived from grains or legumes or roots (e.g., from taro, banana, jackfruit, konjac, lentil, fava, lupin bean, pea, bean, rice, wheat, barley, rye, corn, sweet rice, soy, teff, buckwheat, amaranth, chickpea, sorghum, almond, chia seed, flaxseed, potato, tapioca, potato), protein isolates (e.g., from potato, soy, pea, lentil, chickpea, lupin, oat, canola, wheat), hydrolyzed protein isolates (e.g., hydrolyzed pea protein isolate, hydrolyzed soy protein isolate), protein concentrates (e.g. from algae, lentil, pea, soy, chickpea, rice, hemp, fava bean, pigeon pea, cowpea, vital wheat gluten), beta-glucans (e.g., from bacteria [e.g., curdlan], oat, rye, wheat, yeast, barley, algae, mushroom), gums (e.g., xanthan gum, guar gum, locust bean gum, gellan gum, gum arabic, vegetable gum, tara gum, tragacanth gum, konjac gum, fenugreek gum, gum karaya, gellan gum, high-acetyl gellan gum, lowacetyl gellan gum), native or relatively folded (i.e., not fully in the native functional state but not fully denatured) proteins (e.g., fava protein, lentil protein, pea protein, ribulose-1,5- bisphosphate carboxylase/oxygenase, chickpea protein, mung bean protein, pigeon pea protein, lupin bean protein, soybean protein, white bean protein, black bean protein, navy bean protein, adzuki bean protein, sunflower seed protein), polysaccharides and modified polysaccharides (e.g., methylcellulose, hydroxypropyl methylcellulose, carboxymethyl cellulose, maltodextrin, carrageenan and its salts, alginic acid and its salts, agar, agarose, agaropectin, pectin, alginate), nut and seed butters (e.g., almond butter, cashew butter, hazelnut butter, macadamia nut butter, peanut butter, pecan butter, pistachio butter, walnut butter, pumpkin seed butter, sesame seed butter, soybean butter, sunflower seed butter), enzymes (e.g., trans-glutaminase, thio-oxidoreductase), prolamin proteins (e.g., Zein protein), gelatin, egg protein, potato flakes, okra, tubers, fibers (e.g., psyllium husk), and derivatives and combinations thereof. Examples of suitable stabilizing agents include but are not limited to polymeric biosurfactants, amphipathic polysaccharides (e.g., methylcellulose), lipopolysaccharides, proteins (e.g., pea protein, soy protein, chickpea protein, algae protein, yeast protein, potato protein, lentil protein), or mannoprotein. Non-limiting examples of flavoring agents are animal meat flavor, an animal meat oil, spice extracts, spice oils, natural smoke solutions, natural smoke extracts, yeast extract, and shiitake extract. Additional flavoring agents may include onion flavor, garlic flavor, or herb flavors. Herbs that may be added include basil, celery leaves, chervil, chives, cilantro, parsley, oregano, tarragon, and thyme. Nonlimiting examples of flavor enhancers include glucose, fructose, ribose, arabinose, glucose-6- phosphate, fructose-6-phosphate, fructose- 1,6-diphosphate, inositol, maltose, sucrose, maltodextrin, glycogen, sugars associated with nucleotides, molasses, animal meat flavor, an animal meat oil, spice extracts, spice oils, natural smoke solutions, natural smoke extracts, yeast extract, and shiitake extract. Additional flavoring agents may include onion flavor, garlic flavor, or herb flavors. Herbs that may be added include basil, celery leaves, chervil, chives, cilantro, parsley, oregano, tarragon, and thyme or mixtures thereof. Non-limiting examples of dietary fiber component may include vegetable fibers from carrots, bamboo, peas, broccoli, potatoes, sweet potatoes, corn, whole grains, alfalfa, collard greens, celery, celery root, parsley, cabbage, squash, green beans, common beans, black beans, red beans, white beans, beets, cauliflower, nuts, apple peels, oats, wheat or plantain, or mixtures thereof. Non-limiting examples of dietary fibers including but not limited to pea fiber, oat fiber, bamboo fiber, rice bran, waxy maize, bean fiber, beet fiber, guar gum, pectin, carrageenan, apple fiber, citrus fiber, carrot fiber, barley fiber, psyllium husk, soy fiber, sesame flour, flaxseed fiber, nuts, garcinia fiber, chicory fiber, and fenugreek fiber and combinations thereof. Non-limiting examples of vitamins that can be used include Vitamins A, C, and E. Non-limiting examples of minerals that may be added include the salts of aluminum, ammonium, calcium, magnesium, and potassium Non-limiting examples of pH regulators include food-grade organic or inorganic buffer (including but not limited to potassium metaphosphate, potassium phosphate, potassium phosphate dibasic anhydrous, potassium phosphate monobasic, potassium phosphate tribasic, potassium phosphate dibasic heptahydrate, monobasic sodium acetate, sodium citrate anhydrous and dihydrate, ammonium hydroxide, sodium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate, calcium carbonate, borate, glycine, Tris, Bis-Tris propane, bicine, HEPES, TESm MOBS, DIPSO, TAPS, triethanolamine (TEA), pyrophosphate, HEPPSO, tricine, hydrazine, glycylclycine, 2-amino-2- methyl- 1,3 -propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, tris(hydroxymethyl)aminomethane (THAM), and other materials known to one of ordinary skill in the art. In some embodiments, the buffer is an alkaline buffer for example: ammonium hydroxide, sodium hydroxide, sodium carbonate, ammonium carbonate, calcium carbonate, glycine, TAPS, pyrophosphate, tricine, hydrazine, glycylclycine, 2-amino-2-methy 1-1,3 - propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, or tris(hydroxymethyl)aminomethane (THAM), but non-buffering agents like malic acid, tartaric acid, succinic acid, lactic acid. Non-limiting examples of preservatives include but are not limited to hydroxybenzoate, nitrite, nitrate, sorbic acid, sodium sorbate, sorbates lactic acid, celery extract, propionic acid, benzoic acid, and sodium propionate.

[0170] A meat-like appearance, taste, or smell may be a beef-like appearance, taste, or smell, a poultry-like appearance, taste, or smell, a seafood-like appearance, taste, or smell, a game-like appearance, taste, or smell, a pork-like appearance, taste, or smell, or any combination thereof. In some embodiments, methods of making food products (e.g., meat replicas) may include combining the compositions herein with non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, or any combination thereof wherein the addition of the composition herein provides a meat like texture to the meat substitute. In some other embodiments, methods of making food products (e.g., meat replicas) may include combining the compositions herein with non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, or any combination thereof wherein the addition of the composition herein provides a fat like (i.e., gel like) texture to the meat substitute. A composition herein added to a food product (e.g., meat replicas) may change texture in response to temperature (e.g., cooking). In some embodiments, a composition herein added to a food product (e.g., meat replicas) may change texture in response to temperature, wherein the change in texture of the composition after heating may be reversed upon cooling of the food product.

[0171] Food products contemplated herein include meat, poultry and seafood analogs comprising as a component the disclosed compositions. Non-limiting examples of food products comprising the compositions disclosed herein include products mimicking ground meat, meatloaf mix, steaks, pinwheels, sausages, salami, jerky, bacon, pork boneless rib meat, chicken cutlets, tenders, drumsticks, or hams, soups or stews. Non-limiting examples of poultry analog food products include vegan chicken, mock chicken, vegan turkey, and compositions mimicking nuggets, cutlets, breasts, slices or strips sourced from chicken, quail, duck, ostrich, turkey, bantam, or geese. Non-limiting examples of seafood analog include fish, clams, oysters, mussels, lobsters, shrimp, crab, echinoderms analogs. The compositions described herein may be formulated to mimic any real meat, poultry, or seafood product, such as ground meat, ground meat patties, ground meat meatballs, meat steaks, meat sausage, meat jerky strips, ground chicken, poultry slices, fish fillets, seafood cutlets, seafood pies, salmon burgers, fish sticks, crab cakes, fish burgers, fish cakes, sushi, chowder, bisques, rolls and seafood stews or any combination thereof. In some embodiments, the compositions described herein may be formed as any such product formed from real beef, poultry, or seafood. The present disclosure expressly contemplates, for example, plant-based food compositions in the form of plant-based beef, which may take the form of a ground beef patty or slider, a ground beef meatbail, a beef sausage or hot dog, a cut of beef, corned beef, or a dried beef strip. The meat alternative formulation described herein may alternatively be prepared in the form taken by other real meat products such as meat (beef, chicken, or turkey) nuggets or strips, meat loaf or meat cake forms, canned seasoned meat, sliced meat, sausage of any size, or processed meats such as salami, bologna, luncheon meat and the like. The meat alternative formulation, after cooking, may provide the color, the flavor, and the texture of cooked meat which is pleasurable and palatable to the consumer.

[0172] In some embodiments, the methods of making food-products may include combining the compositions with various ingredients to be used in meat replicas sold in a form such as “ground meat”, burgers/patties, or other forms, for example comparable to Impossible® Burger (from Impossible™ Foods), Beyond Burger® (from Beyond Meat®), Veggie Chik Patty® (from Morningstar Farms®), and Plant-Based Patties from Good & Gather™. Other examples of poultry, meat and seafood analog products that may include compositions provided herein include products like Veggie Meal Starters® from Morningstar Farms®, such as Veggie CHIK’N Nugget, Veggie Popcorn CHIK’N, Veggie CHIK’N Strips, Veggie Grillers®, Veggie Buffalo, beef analogue products made by Beyond Meat® products such as Beyond Beef® Crumbles, Beyond Beef® Ground Beef, and Beyond Beef® Sausage, or fish analog products made by Good Catch like salmon burgers, fish sticks, fish fillets, crab cakes, fish burgers and fish cakes.

[0173] In some embodiments, the compositions disclosed here may also be incorporated into beverages, liquids, gels, pastes, sauces, powder or cubes, soup or stew bases.

[0174] In some embodiments, the compositions may comprise a composition as disclosed herein mixed with other ingredients to form a non-dairy milk product for example cheese replica, icecreams and yogurt. As used herein, a “cheese substitute” or “cheese replica” can be any non-dairy product that serves a role as food or in food that is commonly served by traditional dairy cheese. A cheese “substitute” or “replica” can be a product that shares visual, olfactory, textural or taste characteristics of cheese such that an ordinary human observer of the product is induced to think of traditional dairy cheese. Non-dairy milk products herein refer to an emulsion comprising proteins and fats or a solution or suspension of proteins, sometimes further comprising other solutes that might include carbohydrates, salts and other small molecules that contribute to flavor, texture, emulsion stability, protein solubility or suspension stability, or its ability to support growth of microbial cultures used in making cheese replicas, yogurt replicas, or other replicas of cultured dairy products. embodiment

[0175] Additionally, methods of making food products (e.g., non-dairy milk products, cheese replicas) may include combining the compositions herein with one or more oils or fats isolated from plant sources, recombinant or synthetic sources. One or more oils or fats isolated from plant sources can be, but are not limited to, triglycerides, monoglycerides, diglycerides, sphingosides, glycolipids, lecithin, lysolecithin, phospholipids such as phosphatidic acids, lysophosphatidic acids, phosphatidyl cholines, phosphatidyl inositols, phosphatidyl ethanolamines, or phosphatidyl serines; sphingolipids such as sphingomyelins or ceramides; sterols such as stigmasterol, sitosterol, campesterol, brassicasterol, sitostanol, campestanol, ergosterol, zymosterol, fecosterol, dinosterol, lanosterol, cholesterol, or episterol; free fatty acids such as palmitoleic acid, palmitic acid, myristic acid, lauric acid, myristoleic acid, caproic acid, capric acid, caprylic acid, pelargonic acid, undecanoic acid, linoleic acid (Cl 8:2), eicosanoic acid (C22:0), arachidonic acid (C20:4), eicosapentanoic acid (C20:5), docosapentaenoic acid (C22:5), docosahexanoic acid (C22:6), erucic acid (C22: l), conjugated linoleic acid, linolenic acid (Cl 8:3), oleic acid (Cl 8: 1), elaidic acid (trans isomer of oleic acid), trans-vaccenic acid (Cl 8:1 trans 11), or conjugated oleic acid; or esters of such fatty acids, including monoacylglyceride esters, diacylglyceride esters, and triacylglyceride esters of such fatty acids. In some embodiments, oils/fats isolated from plant sources can include phospholipids, sterols, lipids, or a combination thereof.

[0176] Methods of making food products may include combining the compositions herein with non-dairy oils or fats to form a non-dairy milk. In some embodiments, methods of making food products may include combining the compositions herein with one or more oils or fats also isolated from plant sources, in a colloidal suspension, solution or emulsion to form the non-dairy milk for making a cheese replica.

[0177] In addition, methods of making food products (e.g., non-dairy milk products, cheese replicas) may include combining the compositions herein with at least one additional ingredient. At least additional ingredient suitable for use herein can include, but is not limited to, additional protein, for example a heme-protein, or soy protein, or combination thereof, a non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, emulsifiers, surfactants, binding agents, emulsifiers, plasticizers, thickeners, surfactants, sugars, coloring agents, binding agents, stabilizing agents, flavor enhancers, flavoring agents, fragrance enhancers, nutritional supplements (e.g. vitamins, minerals, antioxidants), essential oils, pH regulators, preservatives, dietary fibers, gluten, and mixtures thereof.

[0178] In certain embodiments, compositions herein may provide food having at least one characteristic of dairy-based food compositions. Non-limiting examples of non-dairy milk products contemplated herein include milk, yogurt, ice cream, butter, cheese, and the like. In some embodiments, compositions herein may provide food having least one characteristic of dairy-based cheese compositions. In some other embodiments, methods of making food products using the compositions herein can form a non-covalent linked protein network, similar to that formed by casein in natural, dairy-based cheese, which is weakened, but not eliminated, at increased temperatures. In still some other embodiments, methods of making food products (e.g., non-dairy milk products, cheese replicas) using the compositions herein can form a gel-like structure which can form a melted-cheese like structure at higher temperatures. In some embodiments, methods of making food products (e.g., non-dairy milk products, cheese replicas) using the compositions herein can form a gel-like structure which can form a melted-cheese like structure at higher temperatures and return to the gel-like structure upon cooling. In some other embodiments, methods of making food products (e.g., non-dairy milk products, cheese replicas) using the compositions herein can have at least one of the following characteristics of dairy-based cheeses: moisture, hardness, gumminess, cohesiveness, brittleness, adhesiveness, meltability, and/or stretchability.

[0179] Food products contemplated herein (e.g., meat replicas, non-dairy milk products) may comprise about 1% to about 99% (e.g., about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%) of any of the compositions herein by weight. In some embodiments, food products (e.g., meat replicas, non-dairy milk products) may comprise about 1% to about 99% (e.g., about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%) of any of the plant isolated proteins herein by weight. In some other embodiments, food products (e.g., meat replicas, non-dairy milk products) may comprise about 1% to about 99% (e.g., about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%) of any of the pea isolated proteins herein by weight. In still some other embodiments, food products (e.g., meat replicas, non-dairy milk products) may comprise about 1% to about 99% (e.g., about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%) of any of the pea isolated proteins herein by weight and at least one additional isolated protein, wherein the total amount of protein may comprise about 1% to about 99% (e.g., about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%) by weight.

EXAMPLES

[0180] The following examples are included to demonstrate preferred embodiments of the disclosure. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventor to function well in the practice of the present disclosure, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the present disclosure.

Example 1. Methods of Preparing Materials and Solutions and Characterization Thereof [0181] In an exemplary method, materials and solutions were prepared for analysis herein. In brief, two pea protein isolates were used in the methods exemplified herein: PISANE C9 and PURIS Pea 870. The pea protein isolate PISANE C9 was obtained from Cosucra Groupe Warcoing S.A. (Warcoing, Belgium). Pea protein isolate PURIS Pea 870 was purchased from Cargill, Inc. (Minneapolis, USA). Beta (P)-casein reduced micellar casein concentrate, alphas (a_s)-casein, and P-casein reduced micellar casein powder were donated by the Department of Soft Matter Science and Dairy Technology (University of Hohenheim, Stuttgart, Germany). Hydrochloric acid (32%), sodium hydroxide pellets, and tris(hydroxymethyl)aminomethane (THAM) were purchased from Carl Roth GmbH & Co. KG (Karlsruhe, Germany). To measure the protein content of liquid protein samples according to methods disclosed herein, absorbent for liquid samples (DumaS orbXE, C. Gerhardt GmbH & Co. KG, Kbnigswinter, Germany) was used. Double deionized water was used throughout the exemplary methods disclosed herein.

[0182] Aqueous protein solutions with a protein concentration of 10% (w/w) were prepared by dissolving pea protein isolate PISANE C9 (17.6% w/w of powder; Cprotein = 66.9%), pea protein isolate PURISpea 870 (17.8% w/w of powder; Cprotein = 66.0%), a_s-casein (13.9% w/w of powder; Cprotein = 82.2%) and micellar casein powder (17.5% w/w of powder, Cprotein = 67.0%) in water (as separate solutions) and stirred overnight (approximately 8-12 hours) at 25 °C to fully hydrate the proteins. Casein concentrate (51.9% w/w of concentrate, Cprotein = 19.3%) is diluted in water to a protein concentration of 10% (w/w) and stirred for about 1 hour.

[0183] To analyze the nitrogen content of the protein samples prepared in the examples herein, a flash combustion method using Dumatherm DT N Pro (C. Gerhardt GmbH & Co. KG, Kbnigswinter, Germany) was used. In brief, protein solutions (10% w/w) were weighed accurately into tin foils (Dumafoil®, C. Gerhardt GmbH & Co. KG, Kbnigswinter, Germany) containing an absorber (DumaSorbXL, C. Gerhardt GmbH & Co. KG, Kbnigswinter, Germany), combusted with oxygen at 1030 °C, and reduced with copper at 750 °C. The respective release of nitrogen (n = 3) was calculated at total nitrogen (%) by the ingrained software (Dumatherm® Manager, C. Gerhardt GmbH & Co. KG, Kbnigswinter, Germany). The following nitrogen conversion factors were used: A standard factor of N x 6.25 and specific conversion factors of N x 5.36 for pea protein and N x 6.36 for casein.

[0184] To assess the visual appearance of the protein samples prepared in the examples herein, images were taken with an iPhoneSE (Apple Inc., Cupertino, CA, USA) under controlled illumination to check the visual appearance of samples 0 hours, 24 hours, and 48 hours after pH adjustment.

[0185] Pea protein isolate PISANCE C9, pea protein isolate PURIS pea 870, micellar casein powder (diafiltered), a_s-casein, and micellar casein concentrate (diafiltered) were characterized according to the methods described herein, as provided in Table 1 and further shown in Fig. 1. The pea proteins and micellar casein had a protein content of approximately 67%, a_s-casein had a protein content of 82%, micellar casein powder contained 67% protein, and micellar casein concentrate had a protein content of 19% (Table. 1). All proteins characterized in a 10% solution in water (H2O) were in a pH range between 5.4 and 7.4 (Table. 1).

Table 1: Characterization of pea protein and casein powder and concentrate

Example 2. Visual Analysis of Structure Formation in Pea and Micellar Casein Samples [0186] In an exemplary method, visual appearance and consistency of samples were assessed according to the methods described herein. The assessments determine consistency (e.g., gel, liquid) with possible aggregate formation (e.g., with flocs) and phase separation (e.g., 1 -phase, 2- phase).

[0187] For principal screenings, the pea protein isolate and micellar casein (10% w/w protein) solution alone were adjusted to pH 2-11 by using HC1 and NaOH solutions (1 N, 3 N, and 10 N). For protein mixtures, the protein solutions (10% w/w) were mixed at a ratio (r) = 1: 1 to obtain mixtures of 5% (w/w) pea and 5% (w/w) micellar casein with a total protein concentration of 10% (w/w). The samples were stirred for 10 minutes before the pH was adjusted to 2-11 by using HC1 and NaOH solutions (1 N, 3 N, and 10 N). Approximately 500 pL of different concentrations of acid or alkaline solutions were used for a 60 mb sample volume to ensure that the protein concentration remained constant in all samples. The samples were stored for 0 hours and 24 hours at 25 °C in the dark before conducting the sample analyses (See Fig. 2). Fig. 3 shows structure formation in pea and micellar casein samples.

[0188] To assess the effect of varying storage times and varying pH, the pea protein isolate and casein (10% w/w protein) solutions alone were adjusted to pH 7-11 by using HC1 and NaOH solutions (1 N, 3 N, and 10 N). To prepare the protein mixtures, the protein solutions (10% w/w) were mixed at a ratio ® = 1 : 1 to obtain mixtures of 5% (w/w) pea and 5% (w/w) casein with a total protein concentration of 10% (w/w). The solutions alone were stored for 0 hours, 24 hours, and 48 hours at 25 °C in the dark before mixing. The mixing pattern is displayed in Fig. 4. After mixing according to the patterns shown in Fig. 4, the samples were adjusted to pH 7-11 by using HC1 and NaOH solutions (1 N, 3 N, and 10 N). Approximately 500 pL of different concentrations of acid or alkaline solutions were used for a 60 mL sample volume to ensure that the protein concentration remained constant in all samples. The samples were stored for 0 hours and 24 hours at 25 °C in the dark before conducting the sample analyses. As an example, Fig. 5A shows PISANE C9 (10% w/w protein immediately after pH adjustment to pH 11 and Fig. 5B shows the same sample after 24 hours at 25 °C. Figs. 6-11 show structure formation in pea and micellar casein samples.

[0189] In sum, data showed strong gelation of PISANE C9 at pH 11 (i.e., alkaline-induced gelation) after 24 hours. High viscosity of casein solutions under alkaline conditions suggested that anisotropic structures for as-casein powder may occur at pH 11. Finally, data showed a change in precipitation pattern in pea protein-casein mixtures compared to the individual proteins.

[0190] To assess the effect of acid hydrolysis of micellar casein, the casein (10% w/w protein) solution was adjusted to pH 2 by using HQ solution (IO N) and stored in the dark for 24 hours and 48 hours. The prepared pea protein solutions were mixed with the casein solution after being stored for 24 hours and 48 hours at a ratio r = 1 :1 to obtain mixtures of 5% (w/w) pea and 5% (w/w) casein with a total protein concentration of 10% (w/w). After mixing, the samples were adjusted to pH 2-11 by using HC1 and NaOH solutions (1 N, 3 N, and 10 N). Approximately 500 pL of different concentrations of acid or alkaline solutions were used for a 30 mL sample volume to ensure that the protein concentration remained constant in all samples. The samples were stored for 0 hours, 24 hours, and 48 hours at 25 °C in the dark before conducting the sample analyses See Fig. 12).

[0191] Fig. 13 shows visual analysis of casein samples were mixed with an aqueous pea protein isolate solution after the casein samples were stored at pH 2 for 24 hours or 48 hours. The samples with the same pea protein isolate mixed with casein after 24 hours of acid hydrolysis did not differ significantly from the ones with 48 hours of acid hydrolysis. All samples from pH 2 to 5 exhibited a formation of white flakes, which were slightly bigger the lower the pH. Samples in the pH range from 6 to 10 were mostly liquid and smooth without any visible particles. The samples with PISANE C9 exhibited a phase separation at pH 3, 4, 5 and 6 after 24 hours and 48 hours, respectively. The isoelectric point (pl) of caseins is around 4.6 and the pl of major pea proteins at 4.5, presumably resulting in precipitation of the proteins. Without being bound to theory, the difference in solubility between the two pea protein isolates used, may be related to potentially different proportions of pea protein subunits, such as vicilin, legumin and covicilin, which have differing molecular sizes and pl.

[0192] Figs. 14A-14D show phase diagrams of samples with previous acidification of casein at pH 2 for 48 hours of PISANE C9 x micellar casein powder (Fig. 14A), PURIS Pea 870 x micellar casein powder (Fig. 14B), PISANE C9 x micellar casein concentrate (Fig. 14C) and PURIS Pea 870 x micellar casein concentrate (Fig. 14D) at pH 2-11.

[0193] Figs. 15A-15D show phase diagrams of samples with previous homogenization of both protein solutions and acidification of casein at pH 2 for 24 hours and 48 hours, of PISANE C9 x micellar casein powder (Figs. 15A-15B) and PISANE C9 x micellar casein concentrate (Figs. 15C-15D) at pH 2-11.

[0194] In sum, data showed occurrence of flocculation in pea protein-hydrolyzed casein mixtures at acidic pH with only a minor presence of gelation in pea protein-hydrolyzed casein mixtures, if only at acidic pH. Also, homogenization of protein solutions had only minor effects on viscosity and flocculation behavior compared to unhomogenized samples.

[0195] Tables 2-7, provide visual descriptions for each sample prepared as described in this exemplary method.

Table 2: Visual description of aqueous 10% (w/w) solutions of pea protein isolate PISANE

C9, pea protein isolate PURISpea 870, micellar casein and alpha S casein.

Table 3: Visual description of aqueous 10% (w/w) mixture of pea protein isolate PISANE C9 (5% (w/w)) and micellar casein (5% (w/w))

Table 4: Visual description of aqueous 10% (w/w) mixture of pea protein isolate PURISpea 870 (5% (w/w)) and micellar casein (5% (w/w)).

Table 5: Visual description of aqueous 10% (w/w) mixture of pea protein isolate PISANE C9 (5% (w/w)) and alpha S casein (5% (w/w))

Table 6: Visual description of samples of pea protein isolate PISANE C9 and PURIS Pea 870 with P-casein reduced micellar casein powder (after being stored for 24 h and 48 h at pH2).

Table 7: Visual description of samples of pea protein isolate PISANE C9 and PURIS Pea 870 with P-casein reduced micellar casein concentrate (after being stored for 24h and 48h at pH2).

Example 3. Microscopic Analysis of Structure Formation in Pea and Micellar Casein Samples [0196] In an exemplary method, microscopic appearance and consistency of samples were assessed according to the methods described herein. To assess the optical microscopy of the protein samples prepared in the examples herein, microscopic images were taken with an Axio Scope. Al LED microscope with Axiocam ICc3, with the software AxioVs40 V 4.8.2.0 at a magnification of lOx. Three images were taken per sample. Representative microscopic images of individual protein samples of pea protein isolate PISANE C9, pea protein isolate PURISpea 870, as-casein, micellar casein powder, and casein concentrate are provided in Fig. 22.

[0197] Samples were prepared according to the methods depicted in Figs. 2, 4, and 12. As a first screening, microscopic images were collected of samples of: 1) pea protein (PISANE C9) (10% w/w); 2) a mixture of pea protein (5% w/w) casein concentrate (5% w/w); and 3) casein concentrate (5% w/w) where the total protein content of each sample was 10% w/w. Fig. 16 shows microscopic images collected from each sample at pH 2-11 after 0 hours or 24 hours of storage.

[0198] Next, microscopic images were collected of samples stored for either 0 hours, 24 hours, or 48 hours at pH ranging from 7-11. Fig. 17 shows images from a PISANE C9 pea protein (5% w/w) and cis-casein powder (5% w/w) mixture at varying storage times and varying pHs. Figs. 18 and 19 show images from a PISANE C9 pea protein (5% w/w) and [3-casein reduced micellar powder (5% w/w) mixture at varying storage times and varying pHs.

[0199] Microscopic images were collected of samples stored for either 0 hours, 24 hours, or 48 hours at pH ranging from 7-11 where the casein used in the sample mixtures was hydrolyzed at pH2 before mixing with a pea protein. Within the samples with PISANE C9, samples from pH 7 to 11 exhibited irregular structure formation with rather big aggregates (Fig. 20). As for the samples with PURIS Pea 870, those structures seemed slightly less irregular and smaller (Fig. 21). Below pH 6, the aforementioned flakes in the solution were partially visible on the microscopic images, e.g. the sample Casein 48 hours x PISANE C9 0 hours at pH 4 (Fig. 20) or Casein 24 hours x PURIS Pea 870 0 hours at pH 3 (Fig. 21) after 0 hours, exhibited a structure network.

Example 4. Texture, Rheology, Stretchability, Moisture Content, and Gelling Analysis of Foodstuff Prototypes

[0200] Foodstuff prototypes containing plant protein isolates stored in alkaline buffer solutions are analyzed with respect to texture, rheology, stretchability and moisture content. The prototypes produced contain the plant protein isolates at different percentages, as well as fats, starches, and/or water. All ingredients and methods used are food grade and readily available.

[0201] Rheological Behavior - Temperature sweeps evaluate the melting ability of foodstuff prototype samples, specifically evaluating the ability of the samples to soften and flow as temperature is increased from 5 °C to 100 °C. The melting profiles of each sample are plotted and differences in melting behavior between samples is determined. Amplitude sweeps performed at 50 °C are performed to examine the rheological properties (linear viscoelastic region and the critical or yield strain value) of the samples when the fat content is sufficiently melted. This ensures that the properties analyzed are primarily attributed to the protein content of the samples.

[0202] Texture Profile Analysis (TPA) - TPA involves a double compression of the samples that mimic the action of chewing. This makes the technique highly reliable when it comes to mechanically determining values for different sensory properties of solid or semi-solid foods. In this work, the values for hardness, springiness, chewiness, and gumminess for all samples are compared. Since the texture is relevant at both low and high temperatures, testing at a range of temperatures, such as 5 °C, 50 °C, and 150 °C, allows for direct comparison of both low and high temperature functionality of all samples.

[0203] Extensibility - The ability of foodstuff prototype samples to stretch at high temperatures is a factor in plant-based food acceptability. Many studies have addressed methods to analyze this property, and identified that results depend on rate of heating, method of heating, and rate of application of stress. To control for this variation, all samples are analyzed using microwave heating (e.g., to approximately 65 °C), and stretched (e.g., at a rate of 2.5 mm/s). The stretch or extensibility is evaluated through measurement of force required to pull the sample, as well as the distance required to break the cheese sample. Differences in the extensibility of the samples are visualized in the different profiles obtained as stretching took place.

[0204] Gelling - Gelling properties of foodstuff prototype samples are compared using small deformation rheology. Gelation of protein dispersion is performed in situ in an Paar Physica MCR501 (Anton Paar Ostfildern, Germany) stress-controlled rheometer, using a concentric cylinder setup (inner and outer cylinder are 8.33 and 9.04 mm respectively). The rheometer is equipped with a Peltier heating and cooling device. Protein dispersion is placed in the geometry and a thin layer of paraffin oil was carefully placed on top to prevent evaporation during the experiment. The temperature is raised from 20 °C to 85 °C at 5 °C/min. After 15 minutes holding at 85 °C, the temperature is decreased to 20 °C at -5 °C/min. After reaching 20° C., the system is left to equilibrate at 0.05% strain and 1 Hz for 10 minutes. A frequency sweep was subsequently performed.

Example 5. Manufacturing of Extruded Textured Food Products

[0205] Protein solutions (10% w/w) are mixed at a ratio (r) = 1: 1 to obtain mixtures of 5% (w/w) pea and 5% (w/w) micellar casein with a total protein concentration of 10% (w/w). The samples are stirred before the pH is adjusted to 2-11 by using HC1 and NaOH solutions (1 N, 3 N, and 10 N). The mix is fed to a 40 kg capacity twin-screw extruder with speed of 25 kg/hour. Screw speed of 300 rpm is settled and temperatures profile 60 °C— >175 °C— >130 °C used in six temperature sections. The mass is let shortly to cool through 10 cm long die.

[0206] After extrusion treatment the post extrusion treatment is carried out by moisturizing the texturized food product with water where the share of water to the dry material is between 1: 1.0 to 1:5. The texturized food product is placed in a liquid and fermented or, alternatively, the food product is hydrated, wetted or soaked for between 1-48 hours before further processing. The brewed (or alternatively, hydrated, wetted or soaked) texturized food product is further treated with amylase and processed with high-speed mixing for 1 -60 minutes. After that an additional high pressure-cooking step is performed in an autoclave or in a pressurized cooking device, preferably having a pressure of at least 2 bar, for 10 to 60 minutes (even more preferably, for around 25 minutes or for between 30 to 60 minutes, such as for 35 to 45 minutes). Instead of the high pressure cooking step, the treated food product may be baked or cooked in a baking or cooking step, preferably in an oven or in a steam oven, at a temperature of between 110 and 130° C., most preferably around 121° C.

[0207] This post extrusion treatment further improves pleasant sensory properties of the texturized food products. Although the example above shows the use of twin-screw extruder, it should be understood that extrusion processes are very diverse and manufacturing of extruded textured food products comprising functionalized plant proteins can be prepared via use of any acceptable model of type food processing extruder.

[0208] The foregoing discussion of the disclosure has been presented for purposes of illustration and description. The foregoing is not intended to limit the disclosure to the form or forms disclosed herein. Although the description of the disclosure has included description of one or more embodiments and certain variations and modifications, other variations and modifications are within the scope of the disclosure, e.g., as can be within the skill and knowledge of those in the art, after understanding the present disclosure. It is intended to obtain rights which include alternative embodiments to the extent permitted, including alternate, interchangeable and/or equivalent structures, functions, ranges or steps to those claimed, whether or not such alternate, interchangeable and/or equivalent structures, functions, ranges or steps are disclosed herein, and without intending to publicly dedicate any patentable subject matter.

Claims

CLAIMS What is claimed is:

1. A composition comprising at least one plant protein isolate, wherein the at least one plant protein isolate is stored in an alkaline buffer solution and forms one or more fiber structures analogous to the fiber structures of an animal protein.

2. The composition according to claim 1 , wherein the alkaline buffer solution comprises a pH ranging from about 10 to about 14.

3. The composition according to claim 1, wherein the total concentration of the at least one plant protein isolate comprises at least 1% w/w.

4. The composition according to claim 3, wherein the total concentration of the at least one plant protein isolate comprises 1% w/w to 20% w/w.

5. The composition according to claim 4, wherein the alkaline buffer solution comprises one or more of ammonium hydroxide, sodium hydroxide, sodium carbonate, ammonium carbonate, calcium carbonate, glycine, TAPS, pyrophosphate, tricine, hydrazine, glycylclycine, 2-amino-2-methyl-l,3-propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, and tris(hydroxymethyl)aminomethane (THAM).

6. The composition according to any one of claim 1 , wherein the at least one plant protein isolate is subjected to an alkaline buffer solution for at least 1 hour.

7. The composition according to claim 1 , wherein the at least one plant protein isolate is subjected to an alkaline buffer solution for about 12 hours to about 48 hours.

8. The composition according to claim 7, wherein the at least one plant protein isolate is subjected to an alkaline buffer solution at a temperature of about 4 °C to about 140 °C.

9. The composition according to claim 8, wherein the at least one plant protein isolate is subjected to an alkaline buffer solution at a temperature of about 20 °C to about 30 °C. The composition according to claim 1 , wherein the at least one plant protein isolate comprises a natural protein isolate, a recombinant protein isolate, or any combination thereof. The composition according to claim 1 , wherein the at least one plant protein isolate comprises one or more proteins present in a crude plant material. The composition according to claim 11, wherein the crude plant material comprises vegetables, fruits, seeds, legumes, grains, or any combination thereof. The composition according to claim 11, wherein the crude plant material comprises legumes. The composition according to claim 11, wherein the crude plant material comprises chickpeas, green peas, yellow peas, black eyed peas, pinto beans, kidney beans, black beans, mung beans, soybeans, adzuki beans, fava beans, edamame, green lentils, red lentils, black lentils, lupins, peanuts, or any combination thereof. The composition according to claim 11, wherein the crude plant material comprises yellow peas. The composition according to claim 1, wherein the at least one plant protein isolate subjected to an alkaline buffer solution comprises at least 10% reactive protein. The composition according to claim 1, wherein the at least one plant protein isolate subjected to an alkaline buffer solution forms a plant-based fiber structure. The composition according to claim 17, wherein the plant-based fiber structure comprises a solid structure. The composition according claim 17, wherein the plant-based fiber structure comprises a gel structure. The composition according to claim 17, wherein the plant-based fiber structure comprises about 10% w/w to about 75% w/w moisture content. The composition according to claim 17, wherein the plant-based fiber structure comprises at least one fiber having a diameter of about 1.5 mm to about 8 mm. The composition according to claim 17 , wherein the plant-based fiber structure comprises at least one fiber having a length of at least 2 mm. The composition according to claim 17, wherein the plant-based fiber structure comprises at least one fiber having a length of about 2 mm to about 40 mm. The composition according to claim 23, wherein the plant-based fiber structure is a solid structure that comprises at least one property of an animal-based food product. The composition according to claim 24, wherein the animal-based food product comprises an animal-based meat product, an animal-based dairy product, or any combination thereof. The composition according to claim 24, wherein the plant-based fiber structure is a solid structure with a water dispersion equal to that of an animal-based food product. The composition according to claim 24, wherein the plant-based fiber structure is a solid structure with a tensile strength equal to that of an animal-based food product. The composition according to claim 17, wherein the plant-based fiber structure comprises an anisotropic structure. The composition according to claim 1 , wherein the alkalinization of the at least one plant protein isolate occurs at a pH ranging from about 9 to about 12. The composition according to claim 1, wherein the alkalinization of the at least one plant protein isolate occurs at a pH ranging from about 11 to about 12. The composition according to claim 17, wherein the plant-based fiber structure formed by the at least one plant protein isolate subjected to an alkaline buffer solution does not change form after neutralizing the pH of the composition. The composition according to claim 31, wherein the pH of the composition is neutralized to a pH ranging from about 6 to about 8. The composition of claim 1, further comprising at least one isolated protein, wherein the at least one isolated protein is isolated from an animal source or is prepared recombinantly in one or more non-animal sources. The composition of claim 33, wherein the one or more non-animal sources is selected from a genetically modified plant, a genetically modified bacteria, a genetically modified yeast, or any combination thereof. The composition according to claim 33, wherein the at least one isolated protein comprises one or more of a-casein, as 1 -casein, as2-casein, P-casein, K-casein, and para- K- casein. The composition according to claim 35, wherein the one or more of a-casein, asl-casein, as2-casein, P-casein, K-casein, and para-K-casein is isolated from an animal source. The composition according to claim 35, wherein the one or more of a-casein, asl-casein, as2-casein, P-casein, K-casein, and para-K-casein is prepared recombinantly in one or more non-animal sources. The composition according to claim 33, wherein the at least one plant protein isolate and the at least one isolated protein are cumulatively present in an amount ranging from about 1% w/w to about 30% w/w of the composition. The composition according to claim 33, wherein the at least one plant protein isolate and the at least one isolated protein are cumulatively present in an amount ranging from about 5% w/w to about 15% w/w of the composition. The composition according to claim 33, wherein the weight ratio of the at least one plant protein isolate to the at least one isolated protein ranges from about 1 : 10 to about 10:1. The composition according to claim 33, wherein the weight ratio of the at least one plant protein isolate to the at least one isolated protein ranges from about 1 :2 to about 2: 1. A method of making a plant-based fiber structure, the method comprising incubating at least one plant protein isolate stored in a buffer solution for about 12 hours to about 48 hours, wherein the buffer solution has a pH ranging from about 10 to about 14. The method according to claim 38, wherein the buffer solution has a pH ranging from about 11 to about 12. The method according to claim 38, wherein the least one plant protein isolate is incubated at a temperature of about 20 °C to about 30 °C. The method according to claim 38, wherein the plant-based fiber structure has at least one property of an animal-based food product comprising an animal -based meat product, an animal-based dairy product, or any combination thereof. The method according to claim 41, wherein the least one property of an animal-based food product comprises color, smell, taste, plasticity, breaking strength, mouth feel, or any combination thereof. The method according to claim 38, wherein the least one plant protein isolate comprises a pea protein isolate. The method according to claim 42, wherein the buffer solution comprises one or more of potassium metaphosphate, potassium phosphate, potassium phosphate dibasic anhydrous, potassium phosphate monobasic, potassium phosphate tribasic, potassium phosphate dibasic heptahydrate, monobasic sodium acetate, sodium citrate anhydrous and dihydrate, ammonium hydroxide, sodium hydroxide, sodium bicarbonate, sodium carbonate, ammonium carbonate, calcium carbonate, borate, glycine, Tris, Bis-Tris propane, bicine, HEPES, TESm MOBS, DIPSO, TAPS, triethanolamine (TEA), pyrophosphate, HEPPSO, tricine, hydrazine, glycylclycine, 2-amino-2-methyl- 1,3 -propanediol (AMPD), HEPBS, taurine, methylamine, piperadine, CABS, and tris(hydroxymethyl)aminomethane (THAM). A composition comprising at least one pea protein isolate subjected to an alkaline buffer solution, wherein: the at least one pea protein isolate comprises about 1% w/w to about 20% w/w of the composition; and, the alkaline buffer solution comprises a pH ranging from about 10 to about 14. The composition of claim 49, wherein the at least one plant protein isolate subjected to an alkaline buffer solution has at least one functional property analogous to a functional property of an animal protein. The composition of claim 50, wherein the at least one functional property analogous to a functional property of an animal protein comprises gelation, solubility, thermal stability, emulsification, foamability, or a combination thereof. A composition comprising at least one pea protein isolate subjected to an alkaline buffer solution, and at least one isolated protein, wherein the at least one isolated protein is isolated from an animal source or is prepared recombinantly in one or more non-animal sources, wherein: the at least one pea protein isolate comprises about 1% w/w to about 20% w/w of the composition; the at least one isolated protein comprises about 1% w/w to about 20% w/w of the composition; the alkaline buffer solution comprises a pH ranging from about 10 to about 14, wherein, a. the composition forms one or more fiber structures analogous to the fiber structures of an animal protein, and b. the composition comprises at least one functional property analogous to a functional property of an animal protein selected from the group consisting of gelation, solubility, thermal stability, emulsification, and foamability, or a combination thereof. The composition according to claim 52, wherein the at least one plant protein isolate and the at least one isolated protein are cumulatively present in an amount ranging from about 5% w/w to about 15% w/w of the composition. The composition according to claim 52, wherein the weight ratio of the at least one plant protein isolate to the at least one isolated protein ranges from about 1 :2 to about 2: 1. A composition comprising: at least one pea protein isolate subjected to an alkaline buffer solution having a pH ranging from about 10 to about 14; and at least one isolated protein comprising one or more of a-casein, as 1 -casein, as2-casein, 0-casein, K-casein, and para-K-casein; wherein the at least one plant protein isolate and the at least one isolated protein are cumulatively present in an amount ranging from about 1% w/w to about 20% w/w of the composition; wherein the weight ratio of the at least one plant protein isolate to the at least one isolated protein ranges from about 1 :5 to about 5: 1; and wherein the composition forms one or more fiber structures analogous to the fiber structures of an animal protein. A food product comprising: a. a composition according to claim 1 ; and b. a protein isolate selected from a casein protein isolate, a whey protein isolate, or a combination thereof; wherein the pH of the food product ranges from about 6 to about 8.5. The food product of claim 56, wherein the food product is a meat replica selected from a meat, poultry or seafood replica. The food product of claim 56, wherein the food product is a non-dairy milk product replica selected from a milk, yogurt, ice cream, butter, or cheese replica. The food product of claim 56, wherein the food product is a liquid or gel composition, selected from a beverage, stew, sauce, paste, spread, or soup. The food product of claim 56, wherein the at least one plant protein isolate is a pea protein isolate. The food product of claim of claim 56, further comprising one or more of an additional protein, a fat, a non-animal-based fat, non-animal- based matrixes, non-animal-based edible fibrous components, emulsifiers, plasticizers, thickeners, surfactants, sugars, coloring agents, binding agents, stabilizing agents, flavor enhancers, flavoring agents, fragrance enhancers, nutritional supplements (e.g. vitamins, minerals, antioxidants), essential oils, pH regulators, preservatives, dietary fibers, gluten, and mixtures thereof . A food product comprising: a. a composition comprising: i. about 5% w/w of at least one pea protein isolate subjected to an alkaline buffer having pH of about 10 to about 14; and ii. about 5% w/w of at least one isolated protein comprising one or more of a micellar casein comprising a-casein, asl -casein, as2-casem, P-casein, K- casein, para-K-casein or a combination thereof; wherein the combination of the at least one pea protein isolate and at least one isolated protein has an adjusted pH of 6-8.5 and is subjected to an extrusion treatment; b. one or more of a protein, a non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, emulsifiers, surfactants, binding agents, thickeners, flavorants, colorants, preservatives, anti-oxidants, plasticizers, nutritional supplements, pH regulators and buffering agents or any combination thereof. A composition comprising: a. about 5% w/w of at least one pea protein isolate subjected to an alkaline buffer having pH of about 10 to about 14; and b. about 5% w/w of at least one isolated protein comprising one or more of a micellar casein comprising a-casein, asl -casein, as2-casein, P-casein, K-casein, para-K-casein or a combination thereof; wherein the combination of the at least one pea protein isolate and at least one isolated protein has an adjusted pH of 6-8.5 and is subjected to an extrusion treatment. A method of making a food product comprising: a. combining: i. at least one plant protein isolate subjected to an alkaline buffer having pH of about 10 to about 14; and ii. at least one isolated protein comprising one or more of a micellar casein comprising a-casein, asl -casein, as2-casein, P-casein, K-casein, para-K- casein or a combination thereof; to form a protein composition; b. subjecting the protein composition to an extrusion treatment to form an extruded protein composition; c. combining the extruded protein composition with one or more of additional proteins, a non-animal-based fat, non-animal-based matrixes, non-animal-based edible fibrous components, emulsifiers, surfactants, binding agents, thickeners, flavorants, colorants, preservatives, anti-oxidants, plasticizers, nutritional supplements, pH regulators and buffering agents or any combination thereof; wherein the food product has one or more fiber structures analogous to the fiber structures of an animal protein. The method of claim 64, wherein the food product comprises the extruded protein composition is m an amount ranging from about 1% w/w to about 20% w/w. The method of claim 64, the at least one plant protein isolate and the at least one isolated protein in the protein composition are combined in a weight ratio ranging from about 1 : 5 to about 5:1.