WO2022192646A1

WO2022192646A1 - Pea protein compositions and uses thereof

Info

Publication number: WO2022192646A1
Application number: PCT/US2022/019911
Authority: WO
Inventors: Roxanna Shariff; Marjorie WELCHOFF; Mark PANCZYK; Nagul NAGULESWARAN; Felix AKHARUME; Catherine Bomont; Sarah Anderson; Atef EBID; LaKendra SHEPARD; Robert SKORGE; Stella COUTROS-HOFFMANN; Canan OZER
Original assignee: Corn Products Development, Inc.
Priority date: 2021-03-12
Filing date: 2022-03-11
Publication date: 2022-09-15

Abstract

This specification discloses pea protein isolates having protein content greater than about 75% (wt.%) and have a molecular weight distribution wherein less than about 40% of the protein in the distribution have molecular weight between about 10 and 30 kDa. Also disclosed are beverages made using the disclosed pea protein isolates. Notably, the beverages good foam stability, stable viscosity and little separation over time.

Description

PEA PROTEIN COMPOSITIONS AND USES THEREOF |00011 The technology disclosed in this specification pertains to pea protein isolates. The disclosed pea protein isolates are useful in food compositions including instant drink mixes and ready -to-drink beverages.

(0002) Plant protein can be isolate from plant organs, like a seeds, to make a powdered ingredient with high plant protein content on a weight basis. Legumes are commonly used to make a plant protein isolate and a commonly used legume is the pea. In one aspect, this specification discloses pea protein isolates having a defined structure. In at least some embodiments the pea protein isolates described in this specification have a molecular weight distribution that generally low amounts of pea proteins within the molecular weight distribution having molecular weight between 10 and 30 kDa. The described pea proteins also generally have high amounts of pea proteins within the molecular weight distribution having a molecular weight about 90 kDa or more preferably about 250kDa.

[(HKD) In another aspect the pea protein isolates described in this specification are useful in food applications, and in at least some preferred embodiments the described pea protein isolates are useful in ready -to-drink beverages and instant beverage mixes. For example, it has been found that the pea protein isolates described in this specification have good foaming qualities and retain formed foams over time. Also, the pea protein isolates described in this specification disperse well in aqueous solution and remain dispersed in solution over time without separating to form a protein poor phase and a protein rich phase.

BRIEF DESCRIPTION OF THE FIGURES

[0004] The technology disclosed in this specification can be better understood with reference to the following figures which are provided for illustrative purposes and are not intended to limit the full scope of the technology disclosed in this specification.

{(HMD) Figure la depicts foam stability and dispersion stability of embodiments of the pea proteins isolates described in this specification.

(0006) Figure lb depicts foam stability and dispersion stability of an alternate pea protein isolates commercial available from Ingredion Incorporated. (0007) This specification discloses pea protein isolates, which in any embodiment described in this specification, have a pea protein in an amount from about 75% to about 90% or from about 75% to about 85%, or from about 80% to about 85%, or from about 80% to about 90%; the pea protein having a molecular weight distribution wherein the percent of protein within the distribution having a molecular weight between about 10 and about 30 kDa (kilodaltons) is less than about 40%, or less than about 35%, or less than about 30%, or less than about 25%, or from about 40% to about 15%, or from about 35% to about 15%, or from about 30% to about 15%, or about 25% to about 15%, or from about 25% to about 20%, or about 30% to about 20% or from about 35% to about 20%, or from about 35% to about 25%, or about 35% to about 30% or about 40% to about 30%.

{0008] In some preferred embodiments described in this specification, a pea protein isolate has pea protein having a molecular weight wherein the percent of protein within the distribution having a molecular between about 10 and about 30 kDa i is less than about 40%. In other preferred embodiments the percent of pea protein in the molecular weight distribution having molecular weight between about 10 and about 30 kDa is from about 30% to about 40%. In still other preferred embodiments the percent of pea protein in the molecular weight distribution having a molecular weight between about 10 and about 30 kDa is from about 30% to about 35%.

|0009| In any embodiment, this specification describes a pea protein isolate having a molecular weight distribution of pea protein wherein the percentage of pea protein within the molecular weight distribution having a molecular weight greater than about 90 kDa is at least about 20%, or at least about 25%, or at least about 26%, or at least about 27%, or at least about 28%, or at least about 29%, or at least about 30%, or from about 20% to about 35%, or from about 25% to about 35%, or from about 26%, or about 27%, or about 28%, or about 29%, or about 30% to about 35%. fOOlO] In any embodiment, this specification describes a pea protein isolate having a molecular weight distribution of pea protein wherein the percentage of pea protein within the molecular weight distribution having a molecular weight from about 90 kDa to about 250 kDa is less than 15%. In some embodiments, percent of pea proteins in the molecular weight distribution having a molecular weight from about 90 kDa to about 250 kDa is from about 10% to about 15%. In some embodiments, the percent of pea proteins in the molecular weight distribution having molecular weight in the range from about 90 kDa to about 250 kDa is less than about 10%, or less than about 5%. In still other embodiments the percent of pea proteins in the molecular weight distribution having a molecular weight in the range from about 1% to about 10%, or more preferably from about 1% to about 5%.

[00111 In any embodiment, this specification describes a pea protein isolate having a molecular weight distribution of pea protein wherein the percentage of pea protein within the molecular weight distribution having a molecular weight greater than about 250 kDa is least about 10%, or at least about 20%. In some preferred embodiments, the percent of pea proteins in the molecular weight distribution having a molecular weight greater than about 250 kDa is at least about 25%, or at least about 26%, or at least about 27%, or at least about 28%, or at least about 29%, or at least about 30%. In still other preferred embodiments the percent of pea proteins in the molecular weight distribution having molecular weight greater than about 250 kDa is from about 20% to about 30%, or from about 21%, or from about 21%, or about 22%, or about 23%, or about 24%, or about 25% to about 30%.

[0012] In any embodiment, this specification describes a pea protein isolate having a particle size distribution having a volume mean diameter of the particles within the distribution from about 40 to about 50 microns, or from about 41 to about 49 microns, or from about 42 to about 48 microns, or about 42 to about 47 microns, or from about 43 to about 47 microns.

[0013] In any embodiment, this specification describes a pea protein isolate having a particle size distribution having a median particle size within the distribution from about 40 to about 50 microns, or from about 41 to about 49 microns, or from about 42 to about 48 microns, or about 42 to about 47 microns, or from about 43 to about 47 microns.

[00.1.4] In any embodiment, this specification describes a pea protein isolate wherein the percent of the protein isolate that is soluble when mixed with water at pH 6 at room temperature is from about 10% to about 17%, or from about 12% to about 17% or from about 14% to about 16%.

[0 15] In any embodiment, this specification describes a pea protein isolate wherein a dispersion of the pea protein isolate dispersed in filtered water (15% to 20% protein to water (w/v)) shows no separation after sitting for 30 minutes. (0016} The pea protein isolates described have a color profile defined by one or more of a Hunter L value of greater than about 80 or greater than about 81, or greater than about 82, or greater than about 83, or from about 80, or about 81, or about 82 or about 83, or about 84 to about 86; a Hunter a value between about 1 and about 3; and a Hunter b value of from about 16 to about 20, or about 18.

(0 17j Pea protein isolates described in this specification can be made by any process useful for obtaining a pea protein isolate having the structure and function described in this specification. In any embodiment, a pea protein isolates described in this specification is made using an isoelectric point separation point process. Isoelectric point separation relies on solubility differences among the components of a flour (e.g. protein, fiber and starch) to isolate and recover protein. An illustrative isoelectric point isolation useful for separating legume proteins and particularly pea proteins follows. Legume protein is highly soluble at pH from about 8 to 10 but starch and fiber are not. So in an aqueous dispersion legume flour is adjusted to pH of about 9, substantial protein is dissolved and is removed as a supernatant or filtrate from the insoluble fiber and starch using filtration or centrifugation. The soluble legume protein is then recovered by adjusting the pH of the solution to the protein’s isoelectric point, generally around pH 4.5, where protein is highly insoluble. A second centrifugation step allows for the protein to be recovered as a precipitate.

|0018| Isoelectric isolation processes tend to damage the protein compared to the protein as seen in flour (not isolated). The degree of damage can be described by the denaturation enthalpy of the protein. In any embodiment described in this specification a pea protein, has denaturation enthalpy of less than about 4 J/g or less than about 1 J/g.

[0019] The pea protein isolates described in this specification are not further modified by enzyme or chemical reaction for example to deamidate or to hydrolyze the protein.

|0020| The pea protein isolates described in this specification are useful in food compositions in an amount from 1% to 99% wt.% of the food composition. The disclosed pea protein isolates are useful to replace animal protein or to provide protein fortification to food products. Illustrative food compositions include gravies, sauces, particularly analog dairy sauces or analog cheese sauces, soups, puddings, salad dressings, analog yogurts, analog sour creams, custards, analog cheese products, baked goods including pastries and dough based baked goods like pie crusts, cookies, breads, crackers, or batter based baked goods like cakes, and muffins. Still other food products include cold pressed nutritional bars. Still other food compositions include ready -to-drink beverages or instant mixes useful to make a beverage.

[00211 The pea protein isolates described in this specification disperse well in water or aqueous solution and provide little viscosity to aqueous solutions and create stable foams in aqueous solutions. Such properties make the disclosed pea protein isolates well suited for use in ready-to- drink beverages or in mixes useful for making a beverage or instant beverage.

[0022] In any embodiment, this specification discloses a composition comprising a pea protein isolate as described in this specification in an amount from about 70% to about 90% (wt.%) of pea protein isolate, or from about 70% to about 85%, or from about 70% to about 80%; and a second ingredient in an amount from about 10% to about 30%, or from about 15% to about 30%, or from about 20% to about 30%. In preferred a food composition is a powdered composition or an instant beverage mix.

[0023] Second ingredients useful in the described food composition include any second ingredient commonly used with a pea protein isolate. Illustrative, non-limiting examples include a prebiotic soluble fibers, sweeteners, flavorings, hydrocolloids, buffering agents and mixtures thereof.

[0024] Prebiotic fibers may include resistant starches, beta-glucans, pectin xylooligosaccharides, fructans and galactans. Preferred prebiotic fibers are soluble in water, for example fructooligosaccharides or short chain fructooligosaccharides. In some embodiments, prebiotic fibers are used in an amount from about 5% to about 15% (wt.%).

[0025] Sweeteners can me caloric or non-caloric and include but are not limited to sucrose, fructose, glucose, allulose, rebaudiosides, steviol glycosides, erythritol, sucralose, and saccharine. In some embodiments, sweeteners are used in amounts from about 0.1% to about 5% (wt.%).

[0026] Flavorings include but are not limited to chocolate, vanilla, fruit, and nut flavorings or other flavorings and spices. In some embodiments, flavorings are used in an amount from about 0.1% to about 5% (wt.%). (0fl27j Hydrocolloids include gums such as xanthan, gellan, guar, acacia, and locust bean. Other common hydrocolloids include agar, carrageenan, modified cellulose (like carboxymethylcellulose or hydropropylcellulose). In some embodiments, hydrocolloids are used in an amount from about 0.1% to about 5% (wt.%);

[0028] Also useful are buffering agents (calcium carbonate, disodium phosphate etc.), salt (Sodium chloride, etc.), which in some embodiments are used in an amount from about 0.1% to about 5% (wt.%). Also useful are added trace elements (iron, manganese, etc.) added in amounts in line with nutritional guidelines.

100291 In some embodiments a composition as described in this specification is a base for a ready -to-drink beverage. In such embodiments, the ready -to-drink beverage comprises water in an amount from about 70% to about 99% (wt.%), or from about 70% to about 95%, or from about, or from about 75% to about 95%, or from about 76% to about 95%, or from about 77% to about 95%, or from about 78% to about 95%, or from about 79%, to about 95%, or from about 80% to about 95%, or from about 80% to about 94%, or from about 80% to about 93%, or from about 80% to about 92%, or from about 80%, to about 91%, or from about 80% to about 90%, or from about 81% to about 90%, or from about 82% to about 90%, or from about to about 90%, or from about 83% to about 84%, or from about 85% to about 90% and the base composition in an amount from about 1% to about 30%, or from about 5% to about 30%, or from about 5% to about 25%, or from about 5% to about 24%, or from about 5% to about 23%, or from about 5% to about 22%, or about 5% to about 21%, or about 5% to about 20%, or about 6% to about 20%, or about 7% to about 20%, or about 8% to about 20%, or about 9% to about 20%, or about 10% to about 20%, or about 10% to about 19%, or about 10% to about 18%, or about 10% to about 17%, or about 10% to about 16%, or about 10% or about 15%.

[0030] In some embodiments a ready-to-drink beverage has a pH in water of from about 6.0 - 8.5. In some embodiments, a composition comprising a pea protein isolate as described in this specification, particularly when used as an instant beverage mix has a pH in water from about 6.0 to about 8.5.

100311 A ready-to-drink beverage, as described in this specification may further comprise edible oils (sunflower oil, soybean oil, coconut oil, alga oil) or other liquid ingredients. (0fl32j In another aspect, this specification discloses a method for making an aqueous solution. In any embodiment described int his specification, an method for making an aqueous solution comprises mixing water and a pea protein isolate or other base composition as described in in this specification to disperse the pea protein isolate or composition in the water; preheating the dispersion of the pea protein isolate or composition in water to a temperature from about 60° to about 70° C; homogenizing the dispersion at pressure of from about 130 to about 140 bar. Optionally, the dispersion is further homogenized at a pressure from about 30 to about 40 bar. Optionally the homogenized dispersion is pasteurizing. An illustrative pasteurization step is to heat homogenized dispersion at a temperature from about 130° to about 140° C for from about 1 and 10 second or from about 5 to about 10 seconds. Follow pasteurization the ready-to-drink beverage is packaged, commonly in sterile containers.

[0033] The pea protein isolates described in this specification have useful function in the described beverages. One useful function is the creation of stable foam in a ready-to-drink beverage or an instant beverage made using a powdered mix using automatic frothers or other vigorous mixing or shaking techniques. Foam stability can be measured as a percent difference between initial foam height and final foam height after sitting for 30 minutes. This specification also describes food compositions that are instant beverages made from a powdered mix or ready- to-drink beverage using the pea protein isolates as described in this specification. Such beverages have a percent change in foam volume of less than 50%, meaning that the percent change in the foam height from initial to end height is less than 50%, or less than 45%, or less than 40%, or less than about 35%, or from about 30% to about 50%, or from about 30% to about 45%, or from about 30% to about 40% after sitting for 30 minutes after creating the foam (ambient temperature).

|0034| The pea protein isolates described in this specification are also useful because they provide little viscosity to ready-to-drink beverages and to instant beverages made from a powdered drink mix. Additionally, the described protein isolates provide stable viscosity to a ready-to-drink and to instant beverages made using and a powdered mix. In such beverages stabile viscosity is measured by reference to changes over time in viscosity in the ready-to-drink beverage or in the beverage from instant drink mix. Embodiments of ready-to-drink beverages described in this specification have viscosity changes over two weeks of less than about 20% or less than 10%. (0fl35j Pea protein isolates described in this specification also usefully remain dispersed within an aqueous solution. In any embodiment, this specification describes a pea protein isolate wherein a dispersion of the pea protein isolate dispersed in filtered water (15% to 20% protein to water (w/v)) shows no separation after sitting for 30 minutes.

[0036] The following definitions and comments are useful for interpreting this specification and understanding the technology disclosed within.

[0037] Use of “about” to modify a number is meant to include the number recited plus or minus 10%. Where legally permissible recitation of a value in a claim means about the value. Use of about in a claim or in the specification is not intended to limit the full scope of covered equivalents.

[0038] Recitation of the indefinite article “a” or the definite article “the” is meant to mean one or more unless the context clearly dictates otherwise.

(0039) While certain embodiments have been illustrated and described, a person with ordinary skill in the art, after reading the foregoing specification, can effect changes, substitutions of equivalents and other types of alterations to the methods, and of the present technology. Each aspect and embodiment described above can also have included or incorporated therewith such variations or aspects as disclosed regarding any or all the other aspects and embodiments.

[0040] The present technology is also not to be limited in terms of the aspects described herein, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. It is to be understood that this present technology is not limited to methods, conjugates, reagents, compounds, compositions, labeled compounds or biological systems, which can, of course, vary. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. It is also to be understood that the terminology used herein is for the purpose of describing aspects only and is not intended to be limiting. Thus, it is intended that the specification be considered as exemplary only with the breadth, scope and spirit of the present technology indicated only by the appended claims, definitions therein and any equivalents thereof. No language in the specification should be construed as indicating any non-claimed element as essential.

[Q0411 The embodiments illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claimed technology. Additionally, the phrase “consisting essentially of’ will be understood to include those elements specifically recited and those additional elements that do not materially affect the basic and novel characteristics of the claimed technology. The phrase “consisting of’ excludes any element not specified.

[0042] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the technology. This includes the generic description of the technology with a proviso or negative limitation removing any subject matter from the genus, regardless of whether the excised material is specifically recited herein.

[0043] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member, and each separate value is incorporated into the specification as if it were individually recited herein.

[Q044] The pea protein isolates described in this specification are further described by the following aspects, which are provided for illustrative purposes and are not intended to limit he full scope of the technology disclosed in this specification.

(0045) A pea protein isolate comprising: a pea protein content in an amount from about 75% to about 90% or from about 75% to about 85%, or from about 80% to about 85%, or from about 80% to about 90%; the pea protein isolate having a molecular weight wherein the percent of pea protein within the molecular weight distribution has a molecular weight between about 10 and about 30 kDa is less than about 40%, or less than about 35%, or less than about 30%, or less than about 25%, or from about 40% to about 15%, or from about 35% to about 15%, or from about 30% to about 15%, or about 25% to about 15%, or from about 25% to about 20%, or about 30% to about 20% or from about 35% to about 20%, or from about 35% to about 25%, or about 35% to about 30% or about 40% to about 30%.

(0046) The pea protein isolate of claim 1 wherein the percent of the pea protein isolate within molecular weight distribution has a molecular weight greater than about 90 kDa is at least about 20%, or at least about 25%, or at least about 26%, or at least about 27%, or at least about 28%, or at least about 29%, or at least about 30%, or from about 20% to about 35%, or from about 25% to about 35%, or from about 26%, or about 27%, or about 28%, or about 29%, or about 30% to about 35%.

[0047] The pea protein isolate of claim 1 or 2 wherein the percent of the pea protein isolate within molecular weight distribution has a molecular weight from about 90 kDa to about 250 kDa less than 15%, or is less than about 10%, or less than about 5%, or from about 10% to about 15%, or from about 1% to about 10%, or from about 1% to about 5%.

[0048] The pea protein isolate of any one of claims 1 to 3 wherein the percent of the pea protein isolate within molecular weight distribution has a molecular weight greater than about 250 kDa is least about 10%, or at least about 20%, or at least about 25%, or at least about 26%, or at least about 27%, or at least about 28%, or at least about 29%, or at least about 30%, or from about 20% to about 30%, or from about 21%, or from about 21%, or about 22%, or about 23%, or about 24%, or about 25% to about 30%.

[0049] The pea protein isolate of any one of claims 1 to 4 further having a particle size distribution wherein the volume mean diameter of the particles within the distribution is from about 40 to about 50 microns, or from about 41 to about 49 microns, or from about 42 to about 48 microns, or about 42 to about 47 microns, or from about 43 to about 47 microns.

[0050] The pea protein isolate of any one of claims 1 to 5 further having a particle size distribution wherein the median particle size within the distribution is from about 40 to about 50 microns, or from about 41 to about 49 microns, or from about 42 to about 48 microns, or about 42 to about 47 microns, or from about 43 to about 47 microns.

[0051 ] The pea protein isolate of any one of claims 1 to 6 wherein the percent of the pea protein isolate that is soluble when mixed with pH 6 water at room temperature is from about 10% to about 17%, or from about 12% to about 17% or from about 14% to about 16%.

[0052] The pea protein isolate of any one of claims 1 to 7 having a color profde defined by one or more of: a) a Hunter L value of greater than about 80 or greater than about 81, or greater than about 82, or greater than about 83 , or from about 80, or about 81 , or about 82 or about 83 , or about 84 to about 86; b) a Hunter a value between about 1 and about 3; and c) a Hunter b value of from about 16 to about 20, or about 18.

[0053] The pea protein isolate of any one of claims 1 to 8 wherein a dispersion of the pea protein isolate dispersed in filtered water (15% to 20% protein to water (w/v)) shows no separation after sitting for 30 minutes.

[0054] A composition comprising: a) from about 70% to about 90% (wt.%) of pea protein isolate as described in any foregoing claim, or from about 70% to about 85%, or from about 70% to about 80%; and b) a second ingredient in an amount from about 10% to about 30%, or from about 15% to about 30%, or from about 20% to about 30% wherein, optionally, the second ingredient is selected from the group consisting of: i) a prebiotic soluble fiber in an amount from about 5% to about 15% (wt.%); ii) a sweetener in an amount from about 0.1% to about 5% (wt.%); iii) a flavoring in an amount from about 0.1% to about 5% (wt.%); iv) an hydrocolloid in an amount from about 0.1% to about 5% (wt.%); v) a buffering agent in an amount from about 0.1% to about 5% (wt.%); and vi) mixtures thereof.

[0055] The composition of claim 10 being a powdered composition, wherein, optionally, the composition is an instant drink mix.

[Q056] The composition of claim 10 or 11 forming a base composition of a ready -to-drink beverage wherein the ready-to-drink beverage comprises: a) water in an amount from about the base composition in an amount from about 70% to about 99% (wt.%), or from about 70% to about 95%, or from about, or from about 75% to about 95%, or from about 76% to about 95%, or from about 77% to about 95%, or from about 78% to about 95%, or from about 79%, to about 95%, or from about 80% to about 95%, or from about 80% to about 94%, or from about 80% to about 93%, or from about 80% to about 92%, or from about 80%, to about 91%, or from about 80% to about 90%, or from about 81% to about 90%, or from about 82% to about 90%, or from about to about 90%, or from about 83% to about 84%, or from about 85% to about 90%; and b) the base composition in an amount from about 1% to about 30%, or from about 5% to about 30%, or from about 5% to about 25%, or from about 5% to about 24%, or from about 5% to about 23%, or from about 5% to about 22%, or about 5% to about 21%, or about 5% to about 20%, or about 6% to about 20%, or about 7% to about 20%, or about 8% to about 20%, or about 9% to about 20%, or about 10% to about 20%, or about 10% to about 19%, or about 10% to about 18%, or about 10% to about 17%, or about 10% to about 16%, or about 10% or about 15%.

[0057] The composition of any one of claims 10 to 12 having a pH in water of from about 6.0 - 8.5

[0058] The composition of any one of claims 10 to 13 having viscosity change over two weeks of less than about 20% or less than 10%, or from about 5% to about 20%, or to about 15%.

[0059] A method making an aqueous solutions comprising: a) mixing water and a pea protein isolate or composition as described in any foregoing claim to disperse the pea protein isolate or composition in the water; b) preheating the dispersion of the pea protein isolate or composition in water to a temperature from about 60° to about 70° C; c) homogenizing the dispersion at pressure of from about 130 to about 140 bar, optionally further homogenizing the dispersion at a pressure from about 30 to about 40 bar; d) optionally , heating the homogenized dispersion at a temperature from about 130° to about 140° C for from about 1 and 10 second or from about 5 to about 10 seconds; e) optionally cooling the homogenized dispersion and transferring it to sterile containers.

[0060] Use of a pea protein isolate as described in any foregoing claim in a ready-to-drink beverage or to make an instant beverage.

[00611 The technology described in this specification can be further understood with reference to the following non-limiting examples, which are provided for illustrative purposes and are not intended to limit the full scope of the technology disclosed in this specification.

EXAMPLE 1 -MAKING OF PEA PROTEIN ISOLATE

[0062] Pea proteins isolates having protein content (dry basis) of about 84% were made using the following process. Dehulled and split yellow pea seeds were cleaned to remove the foreign materials and fed to the pea grind mill to produce pea flour. The flour was then mixed with water and the slurry pH is adjusted (pH —9.0) using dilute sodium hydroxide (NaOH). Next, the flour slurry was passed through hydrocyclones to separate starch (underflow) and protein/fiber (overflow). The protein/fiber overflow from the hydrocyclones was sent to decanters for separation of protein from fibers. Protein solution (overflow) from the decanters was pH adjusted (pH —4.5) for precipitation using dilute hydrochloric acid (HC1) and sent to a pre-heating system to reduce initial microbial load of the protein. The fiber (underflow) was pumped to the fiber separation decanter. Concurrently, the pre-heated and precipitated protein solution was pumped to protein separation decanters. The protein cream (underflow) from the protein separation decanters was collected and pH adjusted in the neutralization tanks and was passed through the pasteurizer where high pressure steam is injected into the protein. The product was then injected into a protein spray dryer. Finally, the spray dried pea protein isolate powder was packaged and stored for functional testing as further described in the following Examples.

EXAMPLE 2 - PHYSICAL CHARACTRISTICS OF PEA PROTEIN ISOLATES

[0063] Pea protein isolates made using the process of Example 1 were evaluated for protein distribution according to molecular weight, particle size distribution, solubility, and color profile. The pea protein isolates were measured to have protein content of about 84% (wt.%) (0fl64j Molecular weight distributions were obtained using SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis). The molecular weight distribution was performed under reducing conditions following the Laemmli procedure.

[Q065] Particle size distribution of the pea isolates was measured dry using a Malvern Mastersizer 3000 particle size analyzer. One teaspoon of sample was used for testing. The following measurements were reported are median particle size, surface area mean and volume mean diameter. Surface area mean diameter or D[3,2] is reported as the diameter of sphere having the mean size within the particle size distribution determined by the volume-to-surface-area ratio of particles in the distribution). Volume mean diameter D[4,3] is reported as the diameter of a particle having the mean particle size in the distribution calculated by reference to the volume proportion of particles within the distribution. Median particle size is reported as the diameter of the median particle.

|0066| Percent protein solubility of a pea protein isolate was determined using a modified method of Morr et al. (J. Food Science 50( 1985) 1715-et seq.) and Karaca et al (Food Res. Inf 1 44(2011) pp. 2742-2750). Protein solutions were prepared by dispersing 1% w/v of protein in buffer with pH adjustment to 6.00 with either 0.1 M NaOH or 0.1 M HC1 as needed. Following establishing desired pH, protein isolate was mixed with solution (solution into protein) by vortexing for 30 sec for 1 hour followed by centrifuging at 4000 x g for 10 min at room temperature. The nitrogen content of the supernatant was determined using LECO protein analyzer (LECO, TruMac® N). Percent protein solubility was calculated by dividing the nitrogen content of the supernatant by the total nitrogen in the sample (x 100%).

Table 1

Physical Attributes of Pea Protein Isolates

[0067] Color profile of pea protein isolates were measured using Hunter Colorimeter. Over various process runs. Color profile of three runs is as follows. One Batch had L = 84, a = 2, and b =17. Another batch had color profile of L = 86.17, a = 1.58, b = 16.50. In still another batch had a color profile ofL = 83.28, a = 2.24 and b = 18.96.

EXAMPLE 3 - FUNCTIONAL OF PEA PROTEIN ISOLATE SOLUTION

[0068] Pea protein isolate function was evaluated in three ways, foaming stability and dispersion stability of a simple solution of pea protein isolate in deionized water, viscosity characteristics of a ready to pea milk, and dispersion stability of a powdered drink mix in an instant beverage.

EXAMPLE 3 A - FOAM STATBILITY

[0069] Foam and stability studies of simple solution consisting of pea protein isolate and deionized were made using Batch 1, Batch 2, Batch 3 and a comparative sample called Comparative Sample 1.

[0070] Comparative Sample l is a pea protein isolate resold by Ingredion Incorporated, having about 80% protein and was measured for molecular weight distribution of the protein composition using SDS-PAGE under reducing conditions following the Laemmli procedure. Comparative Isolate 1 was measured to have about 43% of proteins between about 10 to about 30 kDa and about 21% of proteins having a molecular weight about 90 kDa, all having molecular weight greater than about 250 kDa. It is noted that Comparative Isolate 1 has more protein having molecular weight between about 10 and 30 kDa than Batch 1, 2, or 3 as Comparative Isolate 1 has about 43% of protein within this range but all of Batch 1, 2, and 3 have no more than 38%.

(0071 j Pea protein isolate was mixed with deionized water to form a solution having 5% solids content. Mixing was done by slowly adding filtered water to isolate powder, while stirring. The solution was then covered with parafilm and stirred for 30 min at room temperature (23°C). Protein solution (40 mL) was added to a commercially available automatic milk frother (Capresso) and stirred at “Cold” for 1 min. The solution was then poured into a 250 mL graduated cylinder for evaluation.

(0072] Foam was measured for initial volume. Solution was then left at room temperature for 30 minutes. The foam volume was measured again. Stability of the foam is the percent difference between the initial volume and the final volume. Results are depicted in Figures la and lb for samples made using Batch 3 (Figure la) and Comparative Isolate 1 (Figure lb). Results are discussed in full below in Example 3D. Percent change of foam is not reported numerically, but as seen in Figure lb, for Comparative Isolate 1, the foam volume decreases significantly over 30 minutes sitting time and the dispersion of protein and water separate. Numerical percent change values for dispersions of Batches 1, 2 and 3 of the pea protein isolate in water are as follows. The dispersion using Batch 1 pea protein isolate had percent change of foam volume of 47%. The dispersion using Batch 2 pea protein isolate had percent change of foam volume of 48%. The dispersion using Batch 3 pea protein isolate had percent change of foam volume of 36%. Also notably, with reference to Figure la, the dispersion using Batch 3 does not separate after 30 minutes.

EXAMPLE 3B - READY-TO-DRINK PEA MILK

(0073] Ready-to-drink pea milks were made using the dry formulation set forth in Table 2.

Table 2

Formula for Ready-to-Drink Pea Milk

[0074] Pea protein isolate/gum mix used pea protein isolate from Batch 2 and is a proprietary blend available from Ingredion Incorporated. Ready-to-drink pea milk was made as follows. All ingredients were mixed using a Likwifier mixer at 400rpm for 10 min at ambient temperature (about 23° C). Hydrated mixture was run through a two-stage homogenizer (Microthermics). First stage homogenization began by preheating the mixture to 149° F (65° C). Mixture was then homogenized at a pressure of 2000 psi (about 137 bar). Mixture was further homogenized in second stage at 500 psi (about 37 bar). Entire homogenization process runs for about 4 minutes, starting from preheat through completion of second homogenization stage, based on operation mode of the homogenizer. Following homogenization the mixture was then heated to a final temperature of 275° F (135° C) for 6 seconds for microbial control. The mixture then was cooled and when it reached about 60° F (about 15° C) was transferred to sterilized bottles. The mixture was then stored at refrigerated temperature between 0° and 4° C.

[0075] Viscosity measurements for the ready-to-drink pea milk were taken using a module compact rheometer from Anton Paar at shear rate of 10 s ¹. Viscosity was measured after holding at refrigerated temperature for 24 hours and 1 week. Samples taken out of the refrigerator for testing and measured for viscosity at room temperature. Viscosity was measured to be 33.27 mPa*S after 24 hours and 39.05 mPa*S after 2-weeks.

EXAMPLE 3C - INSTANT PEA BEVERAGE FROM DRY MIX

[0076] Instant beverage was made from a powdered drink mix having the formula set forth in Table 3. Table 3

Instant Beverage Mix (dry ingredients ingredients)

[0077] Powdered drink mix was made using the pea protein isolate of Batch 3 and Comparative Isolate 1. An instant beverage was made from the powdered drink mix by mixing 39 g of the mixture described in Table 3 with 8 fl. oz (about 236 mL) of water (room temperature) in a shaker bottle for 30 seconds. Samples were and then poured into 3 fluid ounce (about 89 ml) for 30 minutes.

EXAMPLE 3D - RESULTS AND COMMENTS

[0078] Results of foaming test and stability test for pea protein isolate in deionized water and of stability test of the instant beverage made from the powdered drink mix are shown in Figures la and lb. The pictures depict results obtained from foaming and stability and instant beverages made using Batch 3 and Comparative Isolate 1. Figure la shows results obtained using Batch 3 pea protein isolate and Figure lb shows results obtained using Comparative Isolate 1. With reference to Figures la and lb show at the top two test tubes containing the pea protein isolate in deionized water mix and at the bottom a 3 -fluid ounce (about 89 ml) container of the instant beverage made from the powdered drink mix using the formula in Table 3. With reference to the two test tubes, the left test tube is a picture of the mixture of pea protein isolate and deionized water (made as described in Example 3a) immediately after being poured from frother into the test tube. The test tube on the right is of the mixture of pea protein isolate and deionized water after sitting for 30 minutes. (0fl79j As seen, shown by the arrows in Figure lb, the solutions using Comparative Isolate 1 separated. Specifically, the arrows point to discoloration in the right test tube and bottom container showing a protein-poor layer between a top foam layer and bottom protein-rich solution layer. Figure la, in contrast, shows only two layers. There is no protein poor layer between the foam and protein rich layer, showing that the protein of Batch 3 has remained evenly dispersed and has not settled from solution.

Claims

CLAIMS What is claimed is:

1. A pea protein isolate comprising: a pea protein content in an amount from about 75% to about 90% or from about 75% to about 85%, or from about 80% to about 85%, or from about 80% to about 90%; the pea protein isolate having a molecular weight distribution wherein the percent of pea protein within the molecular weight distribution that has a molecular weight between about 10 and about 30 kDa is less than about 40%, or less than about 35%, or less than about 30%, or less than about 25%, or from about 40% to about 15%, or from about 35% to about 15%, or from about 30% to about 15%, or about 25% to about 15%, or from about 25% to about 20%, or about 30% to about 20% or from about 35% to about 20%, or from about 35% to about 25%, or about 35% to about 30% or about 40% to about 30%.

2. The pea protein isolate of claim 1 wherein the percent of the pea protein within the molecular weight distribution that has a molecular weight greater than about 90 kDa is at least about 20%, or at least about 25%, or at least about 26%, or at least about 27%, or at least about 28%, or at least about 29%, or at least about 30%, or from about 20% to about 35%, or from about 25% to about 35%, or from about 26%, or about 27%, or about 28%, or about 29%, or about 30% to about 35%.

3. The pea protein isolate of claim 1 or 2 wherein the percent of the pea protein within the molecular weight distribution that has a molecular weight from about 90 kDa to about 250 kDa less than 15%, or is less than about 10%, or less than about 5%, or from about 10% to about 15%, or from about 1% to about 10%, or from about 1% to about 5%.

4. The pea protein isolate of any one of claims 1 to 3 wherein the percent of the pea protein within the molecular weight distribution that has a molecular weight greater than about 250 kDa is least about 10%, or at least about 20%, or at least about 25%, or at least about 26%, or at least about 27%, or at least about 28%, or at least about 29%, or at least about 30%, or from about 20% to about 30%, or from about 21%, or from about 21%, or about 22%, or about 23%, or about 24%, or about 25% to about 30%.

5. The pea protein isolate of any one of claims 1 to 4 further having a particle size distribution wherein the volume mean diameter of the particles within the distribution is from about 40 to about 50 microns, or from about 41 to about 49 microns, or from about 42 to about 48 microns, or about 42 to about 47 microns, or from about 43 to about 47 microns.

6. The pea protein isolate of any one of claims 1 to 5 further having a particle size distribution wherein the median particle size within the distribution is from about 40 to about 50 microns, or from about 41 to about 49 microns, or from about 42 to about 48 microns, or about 42 to about 47 microns, or from about 43 to about 47 microns.

7. The pea protein isolate of any one of claims 1 to 6 wherein the percent of the pea protein isolate that is soluble when mixed with pH 6 water at room temperature is from about 10% to about 17%, or from about 12% to about 17% or from about 14% to about 16%.

8. The pea protein isolate of any one of claims 1 to 7 having a color profile defined by one or more of: a. a Hunter L value of greater than about 80 or greater than about 81, or greater than about 82, or greater than about 83, or from about 80, or about 81, or about 82 or about 83, or about 84 to about 86; b. a Hunter a value between about 1 and about 3; and c. a Hunter b value of from about 16 to about 20, or about 18.

9. The pea protein isolate of any one of claims 1 to 8 wherein a dispersion of the pea protein isolate dispersed in filtered water (15% to 20% protein to water (w/v)) shows no separation after sitting for 30 minutes.

10. A composition comprising: a. from about 70% to about 90% (wt.%) of pea protein isolate as described in any foregoing claim, or from about 70% to about 85%, or from about 70% to about 80%; and b. a second ingredient in an amount from about 10% to about 30%, or from about 15% to about 30%, or from about 20% to about 30% wherein, optionally, the second ingredient is selected from the group consisting of: i. a prebiotic soluble fiber in an amount from about 5% to about 15%

(wt.%); ii. a sweetener in an amount from about 0.1% to about 5% (wt.%); iii. a flavoring in an amount from about 0.1% to about 5% (wt.%); iv. an hydrocolloid in an amount from about 0.1% to about 5% (wt.%); v. a buffering agent in an amount from about 0.1% to about 5% (wt.%); and vi. mixtures thereof.

11. The composition of claim 10 wherein the composition is a powdered composition, wherein, optionally, the composition is an instant drink mix.

12. The composition of claim 10 or 11 wherein the composition forms a base composition of a ready-to-drink beverage wherein the ready-to-drink beverage comprises: a. water in an amount from about the base composition in an amount from about 70% to about 99% (wt.%), or from about 70% to about 95%, or from about, or from about 75% to about 95%, or from about 76% to about 95%, or from about 77% to about 95%, or from about 78% to about 95%, or from about 79%, to about 95%, or from about 80% to about 95%, or from about 80% to about 94%, or from about 80% to about 93%, or from about 80% to about 92%, or from about 80%, to about 91%, or from about 80% to about 90%, or from about 81% to about 90%, or from about 82% to about 90%, or from about to about 90%, or from about 83% to about 84%, or from about 85% to about 90%; and b. the base composition in an amount from about 1% to about 30%, or from about 5% to about 30%, or from about 5% to about 25%, or from about 5% to about 24%, or from about 5% to about 23%, or from about 5% to about 22%, or about 5% to about 21%, or about 5% to about 20%, or about 6% to about 20%, or about 7% to about 20%, or about 8% to about 20%, or about 9% to about 20%, or about 10% to about 20%, or about 10% to about 19%, or about 10% to about 18%, or about 10% to about 17%, or about 10% to about 16%, or about 10% or about 15%.

13. The composition of any one of claims 10 to 12 having a pH from about 6.0 - 8.5

14. The composition of any one of claims 10 to 13 having viscosity change over two weeks of less than about 20% or less than 10%, or from about 5% to about 20%, or to about 15%.

15. A method making an aqueous solutions comprising: a. mixing water and a pea protein isolate or composition as described in any foregoing claim to disperse the pea protein isolate or composition in the water; b. preheating the dispersion of the pea protein isolate or composition in water to a temperature from about 60° to about 70° C; c. homogenizing the dispersion at pressure of from about 130 to about 140 bar, optionally further homogenizing the dispersion at a pressure from about 30 to about 40 bar; d. optionally , heating the homogenized dispersion at a temperature from about 130° to about 140° C for from about 1 and 10 second or from about 5 to about 10 seconds; e. optionally cooling the homogenized dispersion and transferring it to sterile containers.

16. Use of a pea protein isolate as described in any foregoing claim in a ready-to-drink beverage or to make an instant beverage.