Nutritional formulations containing pea protein isolate

Abstract

The present invention relates to a nutritional formulation comprising pea protein isolate, characterized in that the pea protein isolate comprises between 0.5% and 2% free amino acids and has the following viscosity: ‑1 The shear rate ranged from 13×10 ‑3 Pa.s. to 16×10 ‑ 3 Pa.s., in the 40s ‑1 The shear rate ranged from 10×10 ‑3 Pa.s. to 14×10 ‑3 Pa.s., and in the 600s ‑1 The shear rate ranged from 9.8×10 ‑3 Pa.s. to 14×10 ‑3 Pa.s., having the following solubility: from 30% to 40% in the pH range from 4 to 5 and from 40% to 70% in the pH range from 6 to 8. The invention also relates to the use of this nutritional formulation as a sole protein source or as a food supplement intended for infants, children and / or adults.

Description

[0001] This application is a divisional application of the invention patent application with the application date of January 27, 2017, application number 201780008626.9, and invention name “Nutritional formulation containing pea protein isolate”. Technical Field

[0002] The present invention relates to nutritional formulations comprising pea protein isolate.

[0003] More specifically, the present invention relates to the use of these nutritional formulations:

[0004] - as a beverage, through a powder mix to be reconstituted, especially for dietary nutrition (sports, weight loss),

[0005] - as a ready-to-drink beverage for dietary or clinical nutrition,

[0006] - as a liquid for clinical nutrition (enteral bag or drink),

[0007] - Fermented milk of the yogurt type (stirred yogurt, Greek yogurt, drinking yogurt, etc.),

[0008] - as dairy / plant-based beverages,

[0009] - as a dairy / plant-based creamer (e.g. coffee creamer or "coffee punch"), dessert creamer, ice dessert or sorbet,

[0010] - as cookies, muffins, pancakes or nutrition bars (intended for specialized / weight loss nutrition or sports nutrition),

[0011] - As a protein-rich bread or gluten-free bread,

[0012] - as high-protein cereals obtained by extrusion cooking (including "chips" foods / breakfast cereals / snacks),

[0013] -As cheese. Background Art

[0014] Nutritional powders and liquids

[0015] Nutritional powders and liquids manufactured in pediatrics for infants or adults contain a well-defined selection of nutritional ingredients (carbohydrates, proteins, fats, fibers, vitamins and / or trace elements, etc.).

[0016] Some are used as a sole food source, while others are used as food supplements.

[0017] These nutritional products comprise a powder that can be reconstituted with water or another aqueous liquid into a nutritional solution such as an enteral bag or a ready-to-drink beverage.

[0018] These nutritional formulations in powder and liquid form for ready-to-drink beverages and enteral bags are particularly popular in dietetics, and their use is increasing worldwide.

[0019] Nutritional formulations in powder form are typically prepared by intimately mixing the various powders.

[0020] Nutritional formulations for ready-to-drink or enteral administration are typically prepared by preparing one or two separate solutions, which are then mixed together and then heat treated at room temperature to allow storage for at least 12 months.

[0021] The first solution represents the aqueous phase comprising carbohydrates, proteins, fibers, minerals and water-soluble emulsifiers, and the second solution represents the lipid phase comprising oil and fat-soluble emulsifiers.

[0022] As is known, the addition of this second lipid phase depends on the targeted nutritional formulation.

[0023] These nutritional formulations, in powder and liquid form, are particularly sought after for their supply of protein and their supply of energy nutrients.

[0024] Conventionally, milk proteins are used first.

[0025] However, due to cost and environmental considerations, it is preferred to use plant proteins as an alternative to dairy proteins for protein enrichment in powder mix beverages and ready-to-drink beverages.

[0026] Soy proteins (isolates, hydrolysates) are used in large quantities, but rice, wheat and potato proteins are also used (especially to improve the vegetable taste of the finished product).

[0027] In the context of market product re-vegetation and cost reduction, the development of novel solutions based on pea proteins may be suggested as an alternative to dairy proteins in finished products such as beverages (powder mixes to be reconstituted into dietary nutrition (sports / weight loss) and ready-to-drink beverages for clinical and dietary nutrition) and enteral bags for protein enrichment.

[0028] In this case, pea proteins must meet certain functionalities such as good solubility, low viscosity in solution, good resistance to heat treatment of heat-treated liquids, and also good viscosity stability over time.

[0029] It must also meet the nutritional recommendations suggested by FAO / WHO in terms of amino acid profile and digestibility profile.

[0030] It has now been discovered that when protein extracted from pea is chosen for use as a dry mix in a nutritional powder base, even at very low concentrations, and when attempting to rehydrate the nutritional formulation, the formulation may have an undesirable gritty feel in the mouth that is related to the particle size specifications, solubility and composition of the protein.

[0031] Excessive viscosity of pea protein-containing formulations with a high protein content is also a source of dissatisfaction.

[0032] Therefore, alternative solutions to dairy proteins must unquestionably comply with the favorable sensory and functional properties that dairy proteins naturally fulfil.

[0033] Fermented milks or desserts, such as stirred yogurt, Greek yogurt, and set yogurt

[0034] Yogurt is milk that has been inoculated with lactic acid ferments to thicken it and preserve it longer.

[0035] To be called yogurt, it must contain only two specific ferments, Lactobacillus delbrueckii subsp. Bulgaricus and Streptococcus thermophilus, which give it its flavor and texture, and also provide certain nutritional and health benefits.

[0036] Other fermented milks (with a yogurt texture) have been created in recent years. They may or may not contain these two bacteria, as well as additional strains such as Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium bifidum, B. longum, B. infantis, and B. breve.

[0037] Therefore, yogurt is a good source of probiotics (i.e., live microorganisms) that, when consumed in adequate amounts, can have positive health effects beyond their usual nutritional benefits.

[0038] Whether it’s set, stirred or liquid, the name yogurt sticks because, in fact, it’s its manufacturing that determines its final texture, beyond the conventional definition.

[0039] Therefore, to obtain set yogurt, milk is inoculated directly in tanks.

[0040] On the other hand, in the case of stirred yogurt (also known as "Bulgarian" yogurt), the milk is seeded in a trough and then stirred before being poured into its jugs.

[0041] Finally, the liquid yogurt (also known as drinking yogurt) is stirred and then blended until the right texture is achieved, and poured into bottles.

[0042] However, other types of plain yogurt exist, such as Greek yogurt, which have a thicker texture.

[0043] The fat percentage can also change the texture of yogurt, which can be made based on whole milk, semi-skimmed milk or skimmed milk (a label containing the word "yogurt" alone must indicate yogurt made with semi-skimmed milk).

[0044] In all cases, its shelf life cannot exceed 30 days and must always be stored in a refrigerator between 0°C and 6°C.

[0045] The three main types of yogurt are distinguished as follows:

[0046] ○ Stirred yogurt

[0047] More liquid, it's usually more tart than plain yogurt. It's just the texture that's different. It's also called Bulgarian yogurt—a reference to yogurt's supposed origin and the bacteria Lactobacillus bulgaricus (one of the two ferments involved in turning milk into yogurt). It's made in tanks before being packaged in cans.

[0048] It is particularly suitable for preparing beverages such as lassis, fruit cocktails, etc.

[0049] Greek yogurt

[0050] This extra thick yogurt is either plain, strained (traditionally) or enriched with cream. This incredibly delicious yogurt is essential for tsatsiki and Eastern European dishes, and simply blends with herbs to create a delicious aperitif dip. Served cold, it can be used as a substitute for heavy crème fraîche. ).

[0051] ○Drinking yogurt

[0052] While it exists in plain form, it is often sweetened and flavored and made with stirred yogurt. This type of yogurt was conceived in 1974, allowing teenagers to rediscover the joys of milk by eating yogurt straight from the bottle without a spoon. "Pour-over yogurt" in 950g cartons has also recently been introduced, targeting those who want to combine oatmeal and yogurt for breakfast.

[0053] This low-energy, fat-free yogurt made from skim milk – 52 kcal; whole milk yogurt – 88 kcal – "plain" yogurt is naturally low in fat and carbohydrates, yet contains a considerable amount of protein. It is also a source of micronutrients (especially calcium and phosphorus), as well as vitamins B2, B5, B12, and A. Yogurt, which is 80% water, actively participates in the body's hydration.

[0054] Therefore, regular consumption of yogurt is thought to improve the digestion and absorption of lactose (EFSA opinion of 19 October 2010). Other studies have shown potential benefits for improving diarrhoea in children and the immune system in certain people, such as the elderly.

[0055] However, the consumption of milk is increasingly being criticized and questioned, and more and more people are simply deciding to exclude it from their diet, for example because of lactose intolerance or because of allergy problems.

[0056] Therefore, yogurt solutions based on plant milk have been proposed, as plant milk is easier to digest than cow's milk and is rich in vitamins, minerals and unsaturated fatty acids.

[0057] In the remainder of this specification, for simplicity, the term "yogurt" will continue to be used even if the source of the protein is not dairy (officially, "yogurt" made from ingredients other than fermented milk, dairy ingredients or conventional ferments such as Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus does not have the right to be called yogurt).

[0058] The most commonly used plant-based source is soy. However, although soy milk is high in calcium and protein, it is also difficult to digest; this is why it is not recommended for children.

[0059] Additionally, overconsumption of soy-based products is not recommended, as their health effects can be counterproductive when consumed in large quantities.

[0060] Furthermore, it is generally believed that 70% of global soy production is from GMO sources.

[0061] Milk and dairy beverages; plant-based beverages

[0062] Milk is a non-trivial source of protein of high biological quality. For a long time, animal proteins have been overwhelmingly favored for their excellent nutritional qualities, as they contain all the essential amino acids in sufficient proportions.

[0063] However, certain animal proteins can cause allergic reactions, leading to reactions that can be particularly troublesome or even dangerous in everyday life.

[0064] Dairy allergy is one of the most common allergic reactions. Studies have shown that 65% of people who suffer from food allergies are allergic to milk. The adult form of milk allergy, referred to herein as "dairy allergy," is a reaction of the immune system that produces antibodies against the undesirable food. This allergy is different from cow's milk protein (cow protein) allergy (also known as CMPA), which affects newborns and infants. The clinical manifestations of this allergy are primarily gastrointestinal (50% to 80% of cases), as well as cutaneous (10% to 39% of cases) and respiratory (19% of cases).

[0065] In view of all the above-mentioned disadvantages associated with the consumption of dairy proteins, there is a great deal of interest in the use of alternative proteins (also called alternative proteins), including plant proteins.

[0066] Plant-based milks derived from plant ingredients can be an alternative to animal-derived milk. They overcome and avoid CMPA. They are free of casein, lactose, and cholesterol, are rich in vitamins and minerals, and are also high in essential fatty acids but low in saturated fatty acids. Some also have a considerable fiber content.

[0067] Besides the fact that some plant-based milks are low in calcium, and others are not commercially available due to their plant rarity, it should also be mentioned that some plant-based milks can also cause allergic reactions. This is the case, for example, with plant-based milks prepared from oilseed plants (e.g. soy milk).

[0068] Given all the drawbacks of dairy proteins, as well as the harmful allergenic properties imparted by certain plant proteins, there is a real, hitherto unmet consumer demand for plant-based milks that are undisputed and recognized as harmless and can therefore be consumed by the whole family. Traditional manufacturers are also beginning to look for new protein sources to enrich their products.

[0069] The applicant company has also addressed this research in order to be able to meet the increasing demands of manufacturers and consumers for compositions with advantageous nutritional properties without the disadvantages of some existing compositions. The applicant's research has involved formulating new plant-based milks with undisputed and recognized harmlessness and therefore consumable by the whole family.

[0070] Dairy cream for coffee cream, butter, cheese, Chantilly cream, sauces, cake toppings, and decorations

[0071] Dairy creams are products obtained by condensing milk and containing more than 30% fat, in the form of an emulsion of oil droplets in skimmed milk. They can be used for various purposes, either directly as a consumer product (e.g. as coffee creamer) or as an industrial raw material for the manufacture of other products such as butter, cheese, Chantilly cream, sauces, ice cream, or alternatively as cake toppings and decorations.

[0072] There are various types of cream: crème fraîche, low-fat cream, single cream, double cream, pasteurized cream, etc. These creams differ according to their fat content, storage, and texture.

[0073] Raw cream is the cream obtained from the separation of milk and cream, directly after skimming and without undergoing a pasteurization step. It is liquid and contains 30%-40% fat.

[0074] Pasteurized cream (still liquid) has undergone a pasteurization process. It's heated at 72°C for about 20 seconds to eliminate harmful microorganisms. This cream is particularly well-suited for spreading. As a result, it develops a lighter, larger texture when whipped, allowing for the incorporation of air bubbles. For example, it's ideal for Chantilly cream.

[0075] Some liquid creams sold in stores are described as "extended shelf life treated." They can be stored for weeks in a cool, dry place. To preserve them for such long periods, they have been sterilized or heated using the UHT process. For sterilization, this involves heating the cream at 115°C for 15 to 20 minutes. With the UHT (or ultra-high temperature) process, the cream is heated at 150°C for 2 seconds. The cream is then cooled rapidly, which results in a better preservation of its flavor qualities.

[0076] Once separated from the milk and skimmed, cream is naturally fluid. To give it a thicker texture, it undergoes an inoculation step. Lactic acid ferments are incorporated into the cream, and as it matures, it acquires a thicker texture, as well as a more acidic and richer flavor.

[0077] Along with traditional techniques for obtaining cream from milk, which date back thousands or centuries, techniques for combining or rehydrating cream from dairy ingredients have been developed in the past decade.

[0078] Compared with crème fraîche, these new technologies for rehydrating milk cream offer clear advantages in the industrial process: lower raw material storage costs, greater formulation flexibility, and no seasonal influence on milk composition.

[0079] Reconstituted dairy cream can therefore benefit from the natural image usually attributed to dairy products, as the regulation stipulates that its manufacture uses only dairy ingredients with or without the addition of drinking water and with the same finished product characteristics as cow's milk cream (Codex Alimentarius, 2007).

[0080] Developments in the field of rehydrated dairy creams have opened up new possibilities for the formulation of creams and, in particular, the birth of the concept of vegetable-based creams.

[0081] Vegetable-based creams are products similar to dairy creams, in which the milk fat is replaced by vegetable fat (Food Codex, codex Stan 192, 1995).

[0082] They are formulated starting from well-defined amounts of water, vegetable fat, dairy or vegetable protein, stabilizers, thickeners and low molecular weight emulsifiers.

[0083] Physicochemical parameters such as particle size, rheology, stability and expansibility are the characteristics of greatest interest to manufacturers and researchers in the field of replacing dairy cream with plant-based cream.

[0084] For example, as in any emulsion, the size of the dispersed droplets (particle size) is a key parameter for cream characterization since it has a significant impact firstly on other physicochemical properties such as rheology and stability and secondly on sensory properties such as texture and color of the cream.

[0085] The influence of emulsifier type includes low molecular weight emulsifiers such as monoglycerides, diglycerides and phospholipids, and high molecular weight emulsifiers such as proteins, and also protein / low molecular weight emulsifier interactions.

[0086] It is therefore known that the concentration of lipid emulsifiers also has an effect on the droplet size of creams. In protein-stabilized systems, very high concentrations of lipid emulsifiers may lead to a high increase in the average droplet size due to extensive aggregation of droplets after protein desorption.

[0087] The type of protein used in the formulation may also affect the particle size of the cream. Specifically, under the same emulsification conditions, creams based on protein sources rich in casein (e.g., skimmed milk powder) generally have smaller average droplet diameters than those based on protein sources rich in whey proteins (e.g., whey powder).

[0088] The difference in particle size between creams prepared from the two protein sources (casein or whey protein) is related to the difference in interfacial properties at the oil / water interface (casein has a higher interfacial tension reducing capacity than whey protein).

[0089] Furthermore, the protein concentration in the formulation has an impact on the particle size of the cream. Specifically, it has been shown that for a certain mass fraction of oil, the droplet size decreases with increasing protein concentration until a certain concentration, above which the size change is very small.

[0090] The simultaneous presence of low molecular weight (surfactants) and high molecular weight (proteins) amphiphilic molecules in cream formulations is often reflected in a decrease in droplet size during emulsification. Furthermore, competitive adsorption between surfactants and proteins at the oil / water interface often leads to protein desorption from the droplet surface during ripening, which can result in changes in particle size.

[0091] Finally, it appears that the emulsification conditions, the choice of ingredients used in the formulation (including proteins and lipids), and the temperature have an impact on the final properties of the cream.

[0092] It seems likely that plant-based creams could offer new technological properties. Freeze resistance, which could provide ice cream with significant stability, is one example. They can also demonstrate cook-and-serve or cook-and-refrigerate stability, a considerable advantage since these creams can be used to prepare hot or cold meals.

[0093] While plant-based creams can offer new functionalities and show textural properties comparable to or even more interesting than dairy creams, it remains that they can have sensory deficiencies, especially in their taste and smell, sometimes even after the addition of flavorings (as is the case with soy or pea proteins).

[0094] Therefore, the Applicant Company has conducted research on plant-based creamers (including the field of "non-dairy" coffee creamers) to gain further insights into the impact of their ingredients (such as pea protein) and their interactions (protein-protein, protein-fat, protein-water, etc.) on the final properties of the cream.

[0095] The applicant company has also developed a vegetarian cheese recipe.

[0096] Cheese is a food product generally obtained from coagulated milk or dairy cream, subsequently strained, and then optionally fermented and optionally ripened.

[0097] Cheese is primarily made from cow's milk, but can also be made from the milk of goats, sheep, buffalo, or other mammals. A bacterial culture is typically used to acidify the milk. Enzymes, such as rennet, or alternatives such as acetic acid or vinegar, are then added to form a coagulation and curd the milk and whey.

[0098] It is known practice to prepare vegetarian alternatives to cheese, in particular mozzarella-type cheese, by replacing milk caseinates with native starches and modified starches, more particularly acetate-stabilized starches.

[0099] However, attempts are still being made to improve shreddability, melting properties, freeze / thaw stability and taste (especially for pizza preparations in the United States).

[0100] Combinations of oil, modified starch and pea protein were tested but were not entirely satisfactory.

[0101] The Applicant Company has found that the use of pea protein isolate according to the invention makes it possible to meet these specifications, in particular with regard to cuttability, melting and taste.

[0102] ice cream

[0103] Ice cream usually contains animal or vegetable fat, protein (lactoprotein, ovoprotein) and / or lactose.

[0104] In addition to giving ice cream its flavor, the protein also acts as a texturizing agent.

[0105] They are basically produced by weighing the ingredients, pre-mixing them, homogenizing, pasteurizing and freezing them at 4°C (to ripen), and subsequently freezing before packaging and storage.

[0106] However, many people cannot tolerate dairy products or other ingredients of animal origin, which prevents them from consuming milk or regular ice cream.

[0107] For this category of consumers, there has hitherto been no alternative to ice cream containing milk with comparable organoleptic value.

[0108] In the hitherto known ice cream products comprising vegetable ingredients (mainly based on soy), attempts have been made to replace animal emulsifiers with vegetable proteins.

[0109] Dried vegetable proteins are frequently used which are obtained in the course of conventional aqueous or hydroalcoholic extraction and which are obtained in powder form after drying.

[0110] These proteins are shown to be heterogeneous mixtures of polypeptides, certain fractions of which have, to varying degrees, particularly advantageous properties such as emulsifiers or gel formers, eg water-binding agents, foaming agents or texture improvers.

[0111] To date, plant protein products have been obtained almost exclusively from soybeans without fractionation based on their specific functional properties.

[0112] Furthermore, the taste of ice cream prepared with the soy protein is unpleasant.

[0113] The applicant company therefore conducted research on plant-based creams and found that the pea protein isolate according to the invention made it possible to meet the required specifications.

[0114] High-protein biscuit products, pastry products, bakery products and cereal products

[0115] In order to obtain the designation "rich in protein", under the current rules it is necessary that the caloric supply related to protein is greater than or equal to 20% of the total energy supply of the finished product.

[0116] This means that for products with a substantial fat content, such as biscuits or cakes (ranging from a minimum of 10% to a maximum of 25%, with an average content of 18% fat), the level of protein incorporation required to achieve this designation is substantial and greater than 20%.

[0117] However, replacing at least one fifth of a formulation with protein, regardless of the protein and the matrix (biscuit / cake), is a real technical challenge, as these rehydrations have the following consequences:

[0118] - the high protein product produced thereby (in particular related to its hydration level) and also the structural and / or textural properties of the finished product,

[0119] - the process used to manufacture high-protein products (ability to be made by molding, "processability"),

[0120] - Organoleptic qualities of high protein preparations and finished products.

[0121] The applicant company has proposed pea protein for increasing the protein content of biscuits BF, while limiting the negative impact on the workpiece and finished product.

[0122] The solution is derived from pea protein, which has little or no functional properties (emulsifying / gelling ability) and has minimal interaction with water; this protein is sparingly soluble.

[0123] However, this protein does not make it possible to fully satisfy the above-mentioned technical problems.

[0124] Thus, good results may be obtained with a "protein source" biscuit, ie a biscuit in which protein provides 12% of the total caloric supply.

[0125] However, in the name of "rich in protein", this protein BF has limitations and the product is not optimized in terms of texture, which remains mushy.

[0126] The Applicant Company has therefore continued its work on optimizing the quality of vegetable proteins, in particular from pea, by proposing a novel pea protein isolate according to the invention, which better meets technological challenges such as protein enrichment of bakery products.

[0127] In particular, the pea protein isolate obtained according to the invention makes it possible to combine The benefit of BF is little functionality (emulsifier power / gelling power) but high solubility.

[0128] The Applicant Company has thus discovered that these two properties, hitherto unknown, have never been combined, can be combined to provide a protein source allowing a high protein enrichment without negatively affecting the preparation process or the texture of the product or finished product. Summary of the Invention

[0129] The present invention proposes a novel nutritional formulation comprising pea protein isolate which can fully or partially replace milk or soy protein, has a neutral taste, and has properties suitable for:

[0130] o Mixed powder,

[0131] o Protein-containing (or even protein-enriched or "high protein") ready-to-drink UHT sterilized beverages and

[0132] o Enteral nutrition solutions,

[0133] Among them are the need for low viscosity beverages and improved pea protein solubility, and also in the following items:

[0134] - Fermented milk of the yogurt type (stirred yogurt, Greek yogurt, drinking yogurt, etc.),

[0135] - Dairy / plant-based beverages,

[0136] o Dairy / plant-based creamers (e.g., “coffee punch”), iced desserts, or sorbets,

[0137] The emulsifying ability of the pea protein isolate is meaningful for its use in partially or completely replacing milk protein in the matrix of these dairy products.

[0138] o Vegetarian cheese,

[0139] The addition of the pea protein isolate makes it possible to improve the cutting properties, melting properties and taste of mozzarella-type vegetarian cheese.

[0140] The present invention also provides novel nutritional formulations comprising pea protein isolate having properties suitable for:

[0141] - biscuits, muffins, pancakes or nutrition bars (intended for specialized / weight loss nutrition or sports nutrition),

[0142] - protein-rich or gluten-free bread,

[0143] - High-protein cereals obtained by extrusion cooking (including "chips" foods / breakfast cereals / snacks).

[0144] The present invention also leads to an improvement in the taste of pea protein (reduction of pea aroma components, grassy aroma components) so that it is more neutral in applications / finished products (with high content of protein and standards) using pea protein isolate for partial or total replacement of dairy protein, which is an important property for all types of dairy products, dairy beverages or plant-based beverages, fermented milks of the yogurt type, dairy creams or plant-based creams, etc.

[0145] More precisely, the subject of the present invention is a nutritional formulation comprising a pea protein isolate:

[0146] o Contains between 0.5% and 2% free amino acids,

[0147] ○The viscosity at 20℃ is as follows:

[0148] ■In 10s -1 The shear rate ranged from 11×10 -3 Pa.s. to 18×10 -3 Pa.s.,

[0149] In the 40s -1 The shear rate ranged from 9×10 -3 Pa.s. to 16×10 -3 Pa.s., and

[0150] ■In the 600s -1 The shear rate ranged from 8×10 -3 Pa.s. to 16×10 -3 Pa.s.,

[0151] ○Have the following solubility:

[0152] ■ In the pH range from 4 to 5, the pH range is from 30% to 40%.

[0153] ■ From 40% to 70% in the pH zone from 6 to 8.

[0154] Preferably, the pea protein isolate has a digestibility expressed as coefficient of digestibility utilization (CDU) of between 93.5% and 95%.

[0155] Preferably, the degree of hydrolysis (DH) of the pea protein isolate is between 5% and 10%.

[0156] In particular, according to the SYMPHID test, pea protein isolate appears as a "fast viscosity" protein, reflecting the rapid duodenal assimilation of the isolate's constituent amino acids.

[0157] Preferably, the pea protein isolate has been pasteurized at high temperature for a short time before being dried by atomization.

[0158] In one embodiment of the present invention, the nutritional formulation comprises at least one pea protein isolate and at least one milk protein. When the nutritional formulation is in powder form, the milk protein preferably represents at least 10%, 15%, 20%, 25%, 30%, 40%, 45% or 50% by weight relative to the total weight of the protein.

[0159] In another embodiment of the present invention, the nutritional formulation comprises at least one pea protein isolate, another plant protein, such as soy, rice and / or wheat protein, and at least one dairy protein.

[0160] This pea protein isolate represents:

[0161] o between 40% and 100%, preferably between 50% and 100%, 60%-100%, 70%-100%, 80%-100% or 50%-90% of the total protein in the nutritional formulation in powder form,

[0162] o Between 0.1% and 100% of the total protein of the ready-to-drink beverage for clinical nutrition and weight loss, preferably between 20%-100%, 30%-100%, 40%-100%, 50% and 100%, 60%-100%, 70%-100%, 80%-100% or 50%-90% and 60%-100%, 70%-100%, 80%-100% or 50%-90% and 60%-100% of the total protein of the nutritional formulation.

[0163] ○ Between 52% and 100% of the total protein in ready-to-drink beverages for sports nutrition, 60%-100%, 70%-100%, 80%-100% or 50%-90% of the total protein in nutritional preparations,

[0164] o between 0.1% and 100% of the total protein of fermented milk of the yogurt type, preferably 20%-100%, 30%-100%, 40%-100%, between 50% and 100%, 60%-100%, 70%-100%, 80%-100%, 20%-60%, 30%-50% or 50%-90% of the total protein in the nutritional formulation,

[0165] o between 0.1% and 100% of the total protein of the dairy beverage, preferably between 20%-100%, 30%-100%, 40%-100%, 50% and 100%, 60%-100%, 70%-100%, 80%-100% or 50%-90% of the total protein of the nutritional formulation,

[0166] o between 0.1% and 100% of the total protein of the dairy cream, ice dessert or sorbet, more particularly between 50%-100%, 60%-100%, 70%-100%, 80%-100% or 50%-90% of the total protein of the coffee punch, and between 20%-100%, 30%-100%, 40%-100%, 50%-100% or 40%-90% of the total protein of the dairy cream, ice dessert or sorbet,

[0167] o Between 5% and 100% of the total protein of a cookie, muffin, pancake, or nutritional bar (intended for specialized / weight loss nutrition or sports nutrition), preferably between 20%-100%, 30%-100%, 40%-100%, between 50% and 100%, 60%-100%, 70%-100%, 80%-100%, or 50%-90% of the total protein of the nutritional formulation,

[0168] o between 5% and 100% of the total protein of the protein-enriched bread or gluten-free bread, preferably between 10%-100%, 20%-100%, 30%-100%, 40%-100%, between 50% and 100%, 60%-100%, 70%-100%, 80%-100% or 50%-90% of the total protein of the nutritional formulation,

[0169] o Between 5% and 100% of the total protein of high protein cereals obtained by extrusion cooking (including "chips" foods / breakfast cereals / snacks), preferably between 20%-100%, 30%-100%, 40%-100%, between 50% and 100%, 60%-100%, 70%-100%, 80%-100% or 50%-90% of the total protein of the nutritional formulation.

[0170] For vegetarian cheese, about 5% by weight of pea protein isolate in the recipe is sufficient to improve its technical and sensory characteristics.

[0171] For example, the pea protein isolate according to the present invention may represent, specifically by weight, 0.1%-10%, 10%-20%, 20%-30%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% of the total protein in the nutritional formulation, or any combination of these percentage ranges.

[0172] A subject of the present invention is also a nutritional formulation as described above, for use as a sole protein source or as a food supplement intended for infants, children and / or adults.

[0173] A subject of the present invention is also the use of this nutritional formulation as a sole protein source or as a food supplement intended for infants, children and / or adults.

[0174] Detailed Description of the Invention

[0175] The present invention relates to a nutritional formulation comprising a pea protein isolate according to the invention. The present invention also relates to the isolate according to the invention and in particular to the use of the isolate according to the invention for preparing a nutritional formulation.

[0176] More specifically, the present invention relates to the use of these nutritional formulations as beverages for dietary nutrition (sports, weight loss) through powder mixtures to be reconstituted, as well as for clinical nutrition (oral route or enteral bag) and dietary nutrition as ready-to-drink beverages, wherein a low viscosity of the beverage and an improvement in the solubility of the pea protein are sought.

[0177] The invention also relates to the use of these nutritional formulations as dairy or plant-based beverages, in fermented milks of the yogurt type (stirred yogurt, Greek yogurt or drinking yogurt), and as dairy or plant-based creams, ice desserts or sorbets.

[0178] Finally, the present invention relates to the use of these nutritional formulations as biscuits, muffins, pancakes or nutritional bars (intended for specialized / weight loss nutrition or sports nutrition), as protein-enriched bread or gluten-free bread, as high-protein small cereals obtained by extrusion cooking ("chips"), wherein more particularly a high-protein solution is sought without negatively affecting the preparation process or the texture of the product or the finished product.

[0179] With regard to taste, the Applicant Company has discovered that dry blending of pea proteins in protein-rich nutritional formulations in powdered form produces an undesirable sandy sensation in the mouth of the reconstituted powder that can be reduced or eliminated by implementing a specific pea protein isolate.

[0180] It was also found that incorporating the pea protein isolate of the present invention into the nutritional formula made it possible to improve the taste of pea protein by reducing pea aroma components and vegetable aroma components.

[0181] For the purposes of the present invention, the term "nutritional formulation in powder form" means a formulation in powder form comprising:

[0182] o at least one vegetable protein, and in particular a vegetable protein from peas,

[0183] o Optionally at least one protein of dairy origin, and

[0184] o optionally at least one ingredient of the fat and carbohydrate type,

[0185] It is reconstituted with aqueous liquids and is suitable for oral administration to humans.

[0186] As used herein, unless otherwise indicated, the term "dry blend" refers to the mixing of components or ingredients to form a base nutritional powder, or to the addition of dry components or powder-based ingredients in powder or granular form to form a nutritional formulation in powder form.

[0187] All percentages, parts and ratios used herein are by weight of the total formulation, unless otherwise specified.

[0188] The powdered food formulations of the present invention and corresponding manufacturing processes may comprise, consist of, or consist essentially of the essential components of the present invention as described herein and also any additional or optional components described herein or useful in nutritional formulations.

[0189] The nutritional formulation in powder form of the present invention comprises pea protein isolate.

[0190] The powdered nutritional formulations of the present invention are typically in the form of a flowable or substantially fluid granular composition, or at least a granular composition that can be easily molded and measured using a spoon or other similar device, wherein the composition can be easily reconstituted by the intended user with an aqueous solution (typically water) to form a liquid nutritional formulation for immediate oral or enteral use.

[0191] In this context, "immediate" use generally means within 48 hours, more typically within about 24 hours, and preferably immediately after reconstitution.

[0192] Nutritional formulations in powder form include pea protein isolate, which in certain embodiments may comprise up to 100% of the protein provided.

[0193] Food preparations in powder form can be formulated with sufficient types and amounts of all nutrients to form food supplements, or specialized nutritional preparations intended for use by people following specific diets intended for sports diets and weight loss.

[0194] In one embodiment, the powdered nutritional formulation may be formulated for the following uses:

[0195] o for repairing muscles after intense effort, such as in the case of athletes, or

[0196] o To ensure the maintenance or building of muscle mass in athletes, or

[0197] o As a meal replacement for people who wish to lose weight through a satiety-inducing effect.

[0198] Food formulations in powder form may have a caloric density suitable for the nutritional needs of the end user, although in most cases the reconstituted powder will contain from about 350 to about 400 kcal / 100 ml.

[0199] Food formulations in powder form can have a protein content suitable for the nutritional needs of the end user, although in most cases the reconstituted powder contains from about 20 to about 91 g protein / 100 g, including from about 40 to about 65 g protein / 100 g.

[0200] Thus, the formulation may comprise between 20% and 95% protein, such as between 20%-90%, 30%-80% or 40%-60%, relative to the total weight of the formulation.

[0201] For example, the pea protein isolate according to the present invention may represent 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90%, or 90%-100% of the total protein of the formulation, or any combination of these percentage ranges.

[0202] Furthermore, food formulations in powder form can have a fat content suitable for the nutritional needs of the end user, although in most cases the reconstituted powder contains from about 0.5 to about 13 g / 100 g, including from about 3 to about 7 g / 100 g.

[0203] Thus, the formulation may comprise between 0 and 20% lipid, such as between 0.5%-15%, 1%-10% or 3%-7% (particularly % by weight) relative to the total weight of the formulation.

[0204] The powdered nutritional formulations of the present invention can be packaged and sealed in single-use or multi-use containers and then stored under ambient conditions for up to 36 months or longer, more typically from about 12 months to about 24 months.

[0205] For multiple-use containers, they can be opened and capped for repeated use by the end user, provided that the capped package is then stored under ambient conditions (e.g., avoiding extreme temperatures) and the contents are used within about one or two months.

[0206] Fields of application of the nutritional formulations according to the invention are, in particular:

[0207] oDiet and nutrition (exercise, weight loss),

[0208] o clinical nutrition (in the form of a drink, dessert cream or enteral bag),

[0209] o Dairy products (in the form of yogurt, dairy beverages, dairy cream, ice desserts or sherbet), o High-protein biscuit products, pastry products, bakery products and cereal products.

[0210] In the field of sports, protein is known to be involved in the maintenance and growth of muscle. Protein supply is also important for athletes who are engaged in bodybuilding or muscle strengthening.

[0211] This protein must be balanced according to the amino acid profile and must comply with FAO / WHO recommendations. Its digestibility is an important factor, ranging from rapid to slower digestion, depending on the moment the protein is supplied.

[0212] Ready-to-drink protein drinks or high-protein beverages then allow the body to benefit from a selected protein supply with limited calories.

[0213] These high-protein drinks must:

[0214] -Rich in protein and low in carbohydrates and fat;

[0215] -Has a good taste;

[0216] - Designed to aid weight loss by stimulating fat loss and aiding muscle recovery;

[0217] - Produces a feeling of fullness;

[0218] - Helps fight hunger without added sugar or fat;

[0219] -Has a balanced content of essential amino acids, fiber, vitamins and minerals;

[0220] - is low in calories.

[0221] These ready-to-drink beverages can advantageously be prepared with the pea protein isolate according to the invention. They can also be used as the sole protein source.

[0222] For example, plant-based beverages as milk alternatives contain on average 4.5 to 11 g protein per 100 ml of beverage, preferably around 7 g protein per 100 ml, and are very low in fiber (around 0.5 to 1 g per 100 ml).

[0223] Thus, the beverage may comprise between 1% and 20% protein, such as between 3% and 15% or between 6% and 8%, relative to the total weight of the beverage.

[0224] For example, the pea protein isolate according to the present invention can represent 50%-60%, 60%-70%, 70%-80%, 80%-90%, or 90%-100% of the total protein, or any combination of these percentage ranges. Preferably, it represents at least 52%. Specifically, the pea protein supply is between 52% and 100% of the total protein supply.

[0225] For ready-to-drink beverages, the pea protein supply can range from 0 to 100%, preferably from 0.01% or 0.1% to 100%. For example, the pea protein isolate according to the present invention can represent 0.1%-10%, 10%-20%, 20%-30%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% of the total protein or any combination of these percentage ranges.

[0226] Due to the absence of noticeable vegetable flavour components, this protein source is very suitable for any type of beverage and due to its medium viscosity it can be incorporated at up to 100% without compromising the final taste (although for very high levels the addition of flavourings would be advantageous).

[0227] As mentioned above, in the field of "diet" drinks, i.e. drinks intended for use on a low-calorie diet or intended for weight loss, these protein-based or protein-rich drinks are not only effective for rapid muscle gain. This type of drink is also very advantageous in the context of a weight loss diet based on protein consumption.

[0228] It is understood that diet drinks are ideal for aiding weight loss. More specifically, they can:

[0229] - Provides satiety effect

[0230] - Protects muscle and replenishes the body to avoid regaining weight.

[0231] As for "sports" drinks, these weight loss drinks have:

[0232] o Balanced content of essential amino acids, fiber, vitamins and minerals

[0233] o Reduced sugar, fat and calorie content.

[0234] Therefore, protein-based drinks are indeed very effective for quickly losing a few kilograms. These protein-rich products simply reduce or prevent the hunger of the person who consumes them. For example, by taking such a drink, the user can significantly reduce the amount of food consumed and allow for faster weight loss (in the context of replacing meals for weight control, or replacing the total daily food intake for weight control).

[0235] In clinical nutrition, enteral nutrition is known as a nutritional therapy solution administered through a probe, which is used when the digestive tract is functioning normally and is accessible but when the patient cannot eat normally or in case of severe malnutrition.

[0236] This technology allows nutrients to be delivered directly to the digestive tract. This technology completely or partially replaces conventional oral administration with "complete" nutritional formulations (which provide all the nutrients the body needs).

[0237] These formulations are typically packaged in flexible (PVC) bags and administered via a nasogastric or gastrostomy, nasojejunal, nasoduodenal, or jejunostomy probe.

[0238] These nutrient blends consist of proteins, lipids, carbohydrates, vitamins, and minerals, with or without fiber.

[0239] Several categories are distinguished: polymer (standard) mixtures and semi-basic ("reduced") mixtures, the latter being indicated in rather special cases (short bowel syndrome, exocrine pancreatic insufficiency, etc.):

[0240] polymer mixture

[0241] o Low calorie (0.5-0.75 kcal / ml), normal or high protein, with or without fiber

[0242] o Isocaloric (1 kcal / ml), normal or high protein, with or without fiber

[0243] o High calorie (1.25kcal / ml-1.5kcal / ml), normal or high protein, with or without fiber

[0244] oSpecial preparations (disturbance of blood sugar metabolism, respiratory insufficiency).

[0245] Semi-essential blends are isocaloric or hypercaloric, normal or high protein blends based on medium chain triglycerides and peptides.

[0246] Due to its functional properties, pea protein isolate as a protein source is particularly suitable for this purpose.

[0247] Furthermore, they make it possible to maintain the same properties as milk proteins, but at a lower cost.

[0248] In the field of (total or partial) replacement of dairy proteins in yogurts, dairy drinks, dairy creams, ice creams or sorbets, plant proteins are sought with functional properties equivalent to or even superior to those of dairy proteins.

[0249] In this patent application, the term "functional property" refers to any non-nutritional property that affects the effectiveness of an ingredient in a dairy product.

[0250] These different properties contribute to obtaining the desired final characteristics of the dairy product. Some of these functional properties are solubility, viscosity, foaming properties and emulsifying properties.

[0251] Protein also plays an important role in the sensory properties of the food matrix in which it is used, and there is a real synergy between functional and sensory properties.

[0252] Thus, functional properties or functionality of proteins are physical or physicochemical properties that contribute to the organoleptic qualities of food systems produced during technological transformation, storage or preparation in the home kitchen.

[0253] It should be noted that, regardless of the source of the protein, said protein has an impact on the color, flavor and / or texture of the product. These organoleptic characteristics have a decisive influence on the choices made by consumers and, in this case, they are highly considered by manufacturers.

[0254] The functionality of a protein is the result of its intermolecular interactions with its environment (other molecules, pH, temperature, etc.).

[0255] In this case, it is a question of surface properties, which combine the properties of the protein in its interaction with other polar or nonpolar structures in the liquid or gas phase: this covers properties such as emulsification, foaming, etc.

[0256] The Applicant Company has observed that there is a real, unmet need for nutritional formulations having advantageous functional properties and which can be used as at least partial replacements for milk proteins in the preparation of dairy products.

[0257] Pea protein isolate as a protein source is particularly suitable for this purpose due to its taste-improving properties.

[0258] More specifically, in these specific application areas, namely:

[0259] o Yogurt-type fermented milk (stirred yogurt, Greek yogurt, drinking yogurt, etc.),

[0260] o Dairy / plant-based beverages,

[0261] o Dairy / plant-based creamers (e.g., “coffee punch”), iced desserts, or sorbets,

[0262] The applicant company has discovered that:

[0263] - With respect to “non-dairy coffee infuser,” also known as “non-dairy coffee creamer,” as will be presented below:

[0264] oViscosity ratio of pasteurized emulsion before drying The pea protein type is closer to the milk control, which makes it possible to dry low-viscosity emulsions with high solids content;

[0265] o Flocculation in coffee appears to be less pronounced for pea protein isolate according to the present invention. The difference in the content of pea proteins was significant, but this may be related to their improved solubility at the acidic pH of coffee or through better stability to divalent ions contained in the water used for coffee rehydration.

[0266] Furthermore, optionally, in order to improve the flocculation ability, one may choose to add a buffer such as sodium citrate, a NaCl type salt (salt) that promotes protein solubility, or a divalent ion complexing agent that is more effective than phosphate.

[0267] o Regarding the emulsifying capacity of the isolate for this specific application, it may be advantageous to choose to use additional emulsifiers, such as E472 (monoacetyltartaric acid esters and diacetyltartaric acid esters of mono- and diglycerides of fatty acids) or to vary the concentration of E471, or by adjusting the protein concentration or adjusting the homogenization process.

[0268] - Regarding "stirred yogurt", in the manufacturing formula to be described below,

[0269] o The temperature before homogenization can be between 65℃ and 80℃,

[0270] oHomogenization pressure can range from 150 bar to 250 bar,

[0271] o Pasteurization temperature can be in the following ranges: 80℃-85℃ for 30 minutes to 90℃-95℃ for 5 minutes to 10 minutes,

[0272] o The fermentation temperature may be in the range of 30°C to 45°C, preferably 38°C to 42°C,

[0273] o According to the requirement of “yogurt”, the types of ferments may be expanded to all ferments used in the yogurt industry.

[0274] Regarding the preparation of the yogurt:

[0275] In addition to modified starch and pectin, locust bean gum or guar gum can be used as stabilizers in different proportions.

[0276] o The starch of choice is a modified starch, preferably a starch that is very low in viscosity or even completely soluble, and may be present in a proportion ranging from 2.5% to 5%, preferably from 2.8% to 3.5%, by weight of the total composition.

[0277] o The addition of a dairy flavoring may advantageously be chosen to provide a "dairyier" aroma component or a fruity preparation, although in the case of pea protein, the vegetable aroma component is greatly reduced with the pea protein isolate of the present invention.

[0278] - As regards "drinking yoghurts", the recipe according to the invention is similar to that for "stirred yoghurts", but the amount of protein is much lower than that for stirred yoghurts, the amount of starch is preferably chosen between 1.5% and 2.5% by weight of the total composition, and this starch may or may not be in dissolved form in this matrix.

[0279] In the field of protein enrichment, the caloric supply of protein in baked products may prove complex:

[0280] - In dry products containing fat, such as biscuits, this requires the use of high protein concentrations in the formulation. This has certain implications:

[0281] o Texture of the finished product (increased hardness / loss of crispness, mushy texture)

[0282] o Taste of the finished product (bitter, beany, etc.) and

[0283] o Impact on the manufacturing process (forming issues / dough rheology / water competition with other ingredients, etc.)

[0284] - For moist products such as bread, the incorporation of protein has an impact on dough rheology. Since the added protein competes with the gluten network, the result is a reduced bread volume and a more compact and paste-like texture.

[0285] An even more important application is the production of high-protein crisps, ie obtained by extrusion and intended for inclusion in small cereals in cereal bars or other cereal aggregates such as "clusters" or muesli.

[0286] The search for a protein content greater than 70% in these high-protein chips has resulted in a significant reduction in the proportion of starch in the recipe, which is responsible for the expansion and therefore the crispiness. Without these starches, the high-protein chips are dense and very hard.

[0287] For several years, the functionality of proteins has been studied in order to select proteins that have the least impact on the texture of protein-rich baked products.

[0288] For the applicant company, in this case, pea protein was developed BF because of its low solubility and small interaction with water.

[0289] However, this pea protein does not make it possible to fully satisfy the above-mentioned technical problems.

[0290] Therefore, use The texture of BF's "protein-rich" cookies was not optimized and remained mushy.

[0291] For high protein potato chips, BF also did not make it possible to achieve the desired crispy texture.

[0292] In bread, although the volume of the bread increased after baking, its volume was still much lower than that of the control bread.

[0293] In order to solve these difficulties, the Applicant Company has therefore discovered that the pea protein isolate according to the invention makes it possible to:

[0294] - Improved solubility relative to pea protein,

[0295] -Reduce the viscosity of water relative to pea protein.

[0296] The developed pea protein isolate has high solubility and low viscosity, which constitutes a new combination of properties.

[0297] To this end, the Applicant Company has overcome the technical prejudice that in order to solve the problem of baked products, it is quite necessary to choose pea proteins that have little interaction with water, however, it turns out that soluble but less viscous proteins perform better.

[0298] Properties of Pea Protein Isolate

[0299] The pea protein isolates according to the invention are firstly characterized by their free amino acid content (determined according to standard NFEN ISO 13903:2005).

[0300] The value is between 0.5% and 2%.For example, the value may be between 0.5%-1%, 1%-1.5%, or 1.5%-2%, or any combination of these percentage ranges.

[0301] For comparison purposes, pea protein (e.g. S85F) has a free amino acid content of approximately 0.18%.

[0302] The pea protein isolate has a total protein content expressed as N.6.25 exceeding at least 70% by weight of the dry product, preferably at least 80% by weight, for example between 80% and 99%, between 80% and 95%, between 80% and 90% or between 80% and 85%.

[0303] The pea protein isolate according to the invention is further characterized by

[0304] o its viscosity curve in water at 15% solids and 20°C, determined as a function of shear rate;

[0305] o Their solubility curves in water as a function of pH, preferably at 20°C.

[0306] To determine the viscosity curve in water, the following measurements are required

[0307] o Measurements were made on an aqueous solution of pea protein isolate at 15% solids,

[0308] o Using an AR2000 rheometer from TA Instruments,

[0309] oIt has a concentric cylindrical geometry,

[0310] o 600s in 3 minutes -1 0.6×10 -3 shear rate (log), and

[0311] o Temperature of 20°C (temperature equilibration for 3 minutes before testing).

[0312] The shear rates generated in the rheometer made it possible to simulate the processing conditions to which a solution of pea protein isolate according to the invention might be subjected:

[0313] o Therefore, 1s -1 to 10s -1 The shear rate is characteristic of the beverage at rest (a more viscous product is the spoon texture),

[0314] o From the 40s -1 to 50s -1 The shear rate is the texture in the mouth,

[0315] o 300s -1 -1000s -1 The shear rate is equivalent to the shear force in the product delivery pump.

[0316] Therefore, the pea protein isolate according to the present invention has the following viscosity:

[0317] o in 10s -1 The shear rate is from 11×10 -3 Pa.s. to 18×10 -3 Pa.s., preferably 12×10 -3 Pa.s. to 17×10 -3 Pa.s., even more preferably from 13×10 -3 Pa.s. to 16×10 -3Pa.s.,

[0318] o in the 40s -1 The shear rate is from 9×10 -3 Pa.s. to 16×10 -3 Pa.s., preferably 10×10 -3 Pa.s. to 15×10 -3 Pa.s., even more preferably from 10×10 -3 Pa.s. to 14×10 -3 Pa.s., and

[0319] o in the 600s -1 The shear rate is from 8×10 -3 Pa.s. to 16×10 -3 Pa.s., preferably 9×10 -3 Pa.s. to 15×10 -3 Pa.s., even more preferably from 9.8×10 -3 Pa.s. to 14×10 -3 Pa.s.

[0320] This reflects the remarkable stability of the isolates, regardless of the shear forces to which they are subjected.

[0321] Pea protein isolates were then characterized by their water solubility profile as a function of pH.

[0322] The principle of the method used is as follows, as will be demonstrated in the example section:

[0323] o suspending pea protein isolate at 2.5% by weight in distilled water,

[0324] o Adjust to the desired pH: in this case, use 0.1N NaOH or 0.1N HCl to 3, 4, 5, 6, 7, or 8,

[0325] o Mixing at 1100 rpm for 30 minutes,

[0326] o Centrifuge at 3000 g for 15 minutes,

[0327] oMeasure the solids content of a portion of the supernatant.

[0328] Therefore, the solubility of pea protein isolate is:

[0329] o from 30% to 40% in the pH zone from 4 to 5,

[0330] o from 40% to 70% in the pH zone from 6 to 8,

[0331] This reflects their remarkable solubility within these pH zones.

[0332] For comparison purposes, pea protein (e.g. S85F) has:

[0333] o Solubility from 10% to 15% in the pH range from 4 to 5,

[0334] o Solubility from 20% to 50% in the pH range from 6 to 8.

[0335] Pea protein isolates are also characterized by their overall digestibility profile relative to intact pea protein, and by their digestion kinetics.

[0336] As shown below, the digestibility measured in vivo makes it possible to attribute to the pea protein isolate according to the invention a coefficient of digestibility utilization (CDU) value ranging from 93.5% to 95%.

[0337] To measure the digestion kinetics of pea protein isolate, an in vitro model of dynamic digestion under physiological conditions equivalent to the stomach followed by the small intestine was used (see Example 1, Section 4).

[0338] As shown below, the behavior of the isolates according to the invention in this model shows their original positioning between intact pea proteins (digestion "fast intermediate" type) and whey proteins (digestion "fast" type).

[0339] Pea protein isolate was ultimately characterized as a "rapidly digestible protein" in an in vitro digestibility model.

[0340] To obtain this result, the gastric behavior of five proteins (pea protein, whey protein and sodium caseinate, and two batches of pea protein isolate according to the invention) was evaluated in an in vitro digestion model (see Examples on page 32, Example 1, Section 5).

[0341] The digestion kinetics of proteins are largely determined by gastric residence time and gastric emptying time.

[0342] Viscosity is an important characteristic that determines gastric emptying rate. Therefore, in vitro viscosity measurement under gastric conditions was chosen as a relevant parameter for characterizing proteins.

[0343] The protein preparation is introduced into an in vitro system simulating gastrointestinal digestion, in this case a system developed by the company NIZO (SIMPHYD system, meaning SIMulation of PHYsiological Digestion), such as www.nizo.comAs shown in the brochure entitled "Bioavailability of Your Ingredients" on the website, reference is made to the article published in Appl. Environ. Microbiol. 2007 Jan;73(2):508-15.

[0344] This setup provides an online rheological measurement system for comparing the behavior of test proteins.

[0345] The viscosity profiles were measured over time under gastric pH and enzyme release conditions.

[0346] As described below, when compared to whey protein (classified as a “low viscosity” protein) and sodium caseinate (classified as an “extended high viscosity” protein):

[0347] - Pea proteins exhibit a rapid increase in viscosity during acidification, returning to baseline at pH 2 (“rapid intermediate viscosity” proteins), whereas

[0348] The pea protein isolate according to the invention shows a very slight viscosity increase after acidification, which then decreases within 30 minutes to reach a value similar to that of whey protein ("fast viscosity" protein).

[0349] Based on their in vitro gastric behavior, the pea protein isolates according to the invention are therefore rapidly transported to the duodenum, which results in a rapid assimilation of their amino acids.

[0350] The emulsification properties of pea protein isolate were evaluated in comparison with pea protein and milk protein.

[0351] This was performed via the liquid route using a Malvern Mastersizer 2000E particle size analyzer.

[0352] The measuring principle is based on light scattering.

[0353] The powder was dissolved in water containing azide at 1% by weight with stirring at 750 rpm for 6 hours.

[0354] 4 ml of cooking oil combining four vegetable oils (sunflower, rapeseed, "oléisol" hybrid sunflower, grapeseed) (eg oil Lesieur Isio 4) are added at 1% to 20 ml of protein (or protein isolate).

[0355] The entire mixture was mixed in a homogenizer (Ultra-Turrax) at 13500 rpm for 3 minutes, and the emulsion thus formed was then analyzed with a particle size analyzer to determine the size of its fat globules.

[0356] As shown below, pea protein isolate according to the present invention has better emulsifying properties than milk protein.

[0357] Furthermore, their emulsifying properties, which are comparable to those of caseinates, make them particularly advantageous for the preparation of dry emulsions of the "light coffee" type.

[0358] The present invention relates to a pea protein isolate as described above and its use for preparing a nutritional formulation.

[0359] Preparation of pea protein isolate according to the present invention

[0360] The preparation of the pea protein isolate according to the invention comprises enzymatic or non-enzymatic hydrolysis of pea protein such that the pea protein isolate has a degree of hydrolysis (DH) of between 5% and 10%, preferably between 6% and 8% and even more particularly from 6.5% to 7%.

[0361] In a first embodiment, the hydrolysis is performed using endopeptidases.

[0362] A non-specific endopeptidase is selected that is derived from a strain of Aspergillus, particularly a strain of Aspergillus species or Aspergillus oryzae.

[0363] More particularly the endopeptidase EC 3-4-11 is selected.

[0364] The exact amount of enzyme added to the suspension to obtain the desired characteristics of the pea protein isolate will vary depending on the specific characteristics, such as:

[0365] (1) The enzyme or enzyme system used;

[0366] (2) the desired final degree of hydrolysis; and / or

[0367] (3) Desired molecular weight / final distribution.

[0368] Given that these parameters are known, one skilled in the art can readily determine appropriate conditions for obtaining the desired characteristics of pea protein isolate.

[0369] In a specific embodiment, the starting pea protein used to prepare the pea protein isolate according to the present invention is a pea protein composition as described in patent application WO 2007 / 17572 or prepared via the process described in patent application WO 2007 / 17572 (the teaching of which is incorporated by reference). In a specific embodiment, the starting pea protein composition is produced by Roquette Frères under the trade name S85F marketed as a composition.

[0370] In a preferred embodiment of the present invention, the pea protein suspension is brought to a value of 5 to 20%, in particular from 15 to 20%, by weight of solids.

[0371] The reaction temperature is adjusted to a value between 50 and 60°C, preferably around 55°C.

[0372] As a general rule, the enzyme system or enzyme is added to the suspension in an amount ranging from about 0.3% to 1% weight / volume.

[0373] The hydrolysis reaction is typically carried out for a time required to achieve the desired degree of hydrolysis and / or desired molecular weight profile, in the present case from about 45 minutes to about 2 hours and 30 minutes, preferably about 1 hour.

[0374] Again, the time required for the hydrolysis reaction depends on the characteristics as described above, but can be readily determined by one skilled in the art.

[0375] In other embodiments, the suspension containing pea protein can be hydrolyzed using non-enzymatic means, such as mechanical (physical) and / or chemical hydrolysis, which techniques are also well known in the art.

[0376] Once the pea protein has been hydrolyzed to the desired extent, the hydrolysis reaction is stopped, for example, by inactivating the enzyme, or by other standard means.

[0377] In one embodiment, the inactivation of the enzyme is performed by heat treatment.

[0378] According to established practice, the enzyme preparation may be suitably inactivated by raising the temperature of the incubation suspension to a temperature at which the enzyme becomes inactivated, for example to about 70°C for about 10 minutes.

[0379] The pea protein isolate thus obtained is then treated at high temperature for a short time (HTST) and then pasteurized and optionally concentrated to a solids content of 10% to 30% and then dried by atomization. For example, the isolate can be pasteurized at a temperature between 130° C. and 150° C. for a time of about 1 second to about 30 seconds.

[0380] Therefore, the present invention relates to a pea protein isolate obtained or obtainable via the above-described method.

[0381] The present invention also relates to a nutritional formulation comprising a pea protein isolate according to the invention and also to the use of this isolate for the preparation of a nutritional formulation.

[0382] The pea protein isolate according to the present invention may be present in the nutritional formulation according to the present invention in an amount of up to 100% by weight, in particular in an amount between 52% and 60% by weight of the nutritional formulation. For example, the pea protein isolate according to the present invention may represent 0.1%-10%, 10%-20%, 20%-30%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% of the total protein in the nutritional formulation, or any combination of these percentage ranges.

[0383] Furthermore, the pea protein isolate according to the present invention may represent 0.1%-10%, 10%-20%, 20%-30%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% by weight of the nutritional formulation or any combination of these percentage ranges. Preferably, it represents 0.1%-60%, 1%-50%, 1%-20% or 1%-10% or any combination of these percentage ranges.

[0384] In a specific embodiment, the pea protein isolate according to the present invention can represent 0.1%-10%, 10%-20%, 20%-30%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% by weight of the nutritional formulation, or any combination of these percentage ranges, and it can represent 0.1%-10%, 10%-20%, 20%-30%, 40%-50%, 50%-60%, 60%-70%, 70%-80%, 80%-90% or 90%-100% or any combination of these percentage ranges of the total protein of the nutritional formulation. Preferably, it represents 0.1%-60%, 1%-50%, 1%-20% or 1%-10% or any combination of these percentage ranges.

[0385] At least a portion of the pea protein isolate present in the powdered food formulation is dried by atomization prior to being introduced (by dry mixing or the like) into the powdered nutritional formulation.

[0386] Properties of other ingredients

[0387] The nutritional formulation in powder form may comprise at least one fat, one protein, or one carbohydrate, wherein at least some of the protein is pea protein isolate.

[0388] The liquid nutritional formulation may comprise at least one protein, carbohydrates, and fat, wherein at least some of the protein is pea protein isolate.

[0389] Generally, sources of fat, carbohydrates, and protein may be used herein in addition to pea protein isolate, provided that these macronutrients are also compatible with the essential components of the nutritional formulation according to the present invention.

[0390] While the total concentrations or amounts of fat, protein, and carbohydrates can vary depending on the nutritional needs of the user, these concentrations or amounts generally fall within one of the following ranges, including any other essential fats, proteins, carbohydrates, and / or ingredients as described herein:

[0391] 1) For powder mixes for beverages:

[0392] o a fat concentration of from about 0.5% to about 13%, preferably from about 1% to about 9%, even more preferably from about 1% to about 3% by weight of the nutritional formulation in powder form;

[0393] o the protein concentration is from about 20% to about 91%, preferably from about 40% to about 90%, even more preferably between about 40% and about 65% by weight of the nutritional formulation in powder form;

[0394] o The carbohydrate concentration is from about 0.9% to about 70%, preferably from about 2% to about 7%, even more preferably between about 20% and about 40% by weight of the nutritional formulation in powder form.

[0395] 2) For liquids:

[0396] o the fat concentration is from about 1% to about 10%, preferably from about 1.5% to about 7%, even more preferably from about 1.5% to about 5% by weight of the nutritional formulation in liquid form;

[0397] o protein concentration is from about 1% to about 15%, preferably from about 3% to about 11%, even more preferably from about 4% to about 7% by weight of the nutritional formulation in liquid form;

[0398] o The carbohydrate concentration is from about 5% to about 45%, preferably from about 9% to about 20%, even more preferably from about 13% to about 17% by weight of the nutritional formulation in liquid form.

[0399] 3) For use in dairy products (in the form of yogurt, dairy beverages, dairy cream, ice desserts or sorbet):

[0400] o a fat concentration of from about 0% to about 15%, preferably from about 1.5% to about 10%, even more preferably from about 3% to about 6% by weight of the nutritional formulation in liquid form;

[0401] o the protein concentration is from about 1% to about 25%, preferably from about 2% to about 20%, even more preferably from about 2.5% to about 15% by weight of the nutritional formulation in liquid form;

[0402] o The carbohydrate concentration is from about 5% to about 45%, preferably from about 9% to about 25%, even more preferably from about 13% to about 20% by weight of the nutritional formulation in liquid form.

[0403] 4) Used in high-protein biscuit products, pastry products, bakery products and cereal products:

[0404] o the fat concentration is from about 0% to about 25%, preferably from about 1% to about 20%, even more preferably from about 10% to about 18% by weight of the nutritional formulation in liquid form;

[0405] o protein concentration is from about 1% to about 30%, preferably from about 2% to about 25%, even more preferably from about 2.5% to about 15% by weight of the nutritional formulation in liquid form;

[0406] o The carbohydrate concentration is from about 15% to about 75%, preferably from about 20% to about 60%, even more preferably from about 20% to about 55% by weight of the nutritional formulation in liquid form.

[0407] Non-limiting examples of fats (in powder or liquid form) or suitable sources thereof for use in the powder and liquid food formulations described herein include coconut oil, fractionated coconut oil, soybean oil, corn oil, olive oil, safflower oil, oleic acid-enriched safflower oil, sunflower oil, oleic acid-enriched sunflower oil, palm and palm kernel oils, palm olein, canola oil, marine crude oil, cottonseed oil, dairy-derived fats, and combinations thereof.

[0408] Non-limiting examples of carbohydrates or suitable sources thereof for use in the powder and liquid forms of the food products described herein can include maltodextrin, dextrin, corn starch or hydrolyzed or modified corn starch, glucose polymers, corn syrup, carbohydrates derived from rice, glucose, fructose, lactose, high fructose syrup, honey, sugar alcohols (e.g., maltitol, erythritol, or sorbitol), and combinations thereof.

[0409] Non-limiting examples of proteins, including pea protein isolate, for use in food products in powder and liquid form include hydrolyzed, partially hydrolyzed, or non-hydrolyzed proteins or protein sources that can be derived from any known source, such as milk (e.g., casein or whey), derived from animals (e.g., meat or fish), derived from cereals (e.g., rice or corn), derived from oil-containing plants (soybeans or rapeseed), legumes with seeds (lentils, chickpeas, or beans), or combinations thereof.

[0410] Non-limiting examples of such proteins include milk protein isolate, milk protein concentrate such as whey protein concentrate, casein, whey protein isolate, caseinates, whole milk, skim milk, soy protein, partially or fully hydrolyzed protein isolate, concentrated soy protein, and the like.

[0411] In a particular embodiment, the nutritional formulation in powder form comprises a combination of pea protein isolate and milk-based protein.

[0412] In one example of this embodiment, the milk-based protein is present in the powdered nutritional formulation in an amount of at least 10%, 15%, 20%, 25%, 30%, 40%, 45% or 50% by weight relative to the total weight of the protein, preferably about 45% by weight relative to the total weight of the protein. For example, the milk-based protein is present in the powdered nutritional formulation in an amount of 10%-60%, 20%-50%, 30%-40% by weight relative to the total weight of the protein. Preferably, the remainder of the protein is provided by the pea protein isolate according to the present invention.

[0413] In another example of this embodiment, the milk-based protein is present in the liquid nutritional formulation for clinical nutrition in an amount of at least 10%, 15%, 20%, 25%, 30%, 40%, 45% or 50% by weight relative to the total weight of the protein, preferably about 50% by weight. For example, the milk-based protein is present in the liquid nutritional formulation for clinical nutrition in an amount of 10%-60%, 20%-50%, 30%-40% or 45%-55% by weight relative to the total weight of the protein. Preferably, the remainder of the protein is provided by the pea protein isolate according to the present invention.

[0414] In another example of this embodiment, the milk-based protein is present in the liquid nutritional formulation for sports in an amount of at least 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60% or 75% by weight relative to the total weight of the protein, preferably about 75% by weight. For example, the milk-based protein is present in the liquid nutritional formulation for sports in an amount of 10%-60%, 20%-50%, 30%-40% or 45%-55% by weight relative to the total weight of the protein. Preferably, the remainder of the protein is provided by the pea protein isolate according to the present invention.

[0415] Nature of optional ingredients

[0416] The nutritional formulation according to the invention may also contain other ingredients which may modify the chemical, physical, hedonic or processing characteristics of the product or serve as a medicament or additional nutritional component when used by certain target groups.

[0417] Many of these optional ingredients are known or otherwise suitable for use in other food products and may also be used in nutritional formulations according to the present invention, provided that the optional ingredients are safe and effective for oral administration and are compatible with the other essential ingredients of the selected product.

[0418] Non-limiting examples of such optional ingredients include preservatives, antioxidants, emulsifiers, buffers, pharmaceutically active agents, other nutrients, dyes, flavorings, thickeners, stabilizers, and the like.

[0419] Nutritional formulations in powder or liquid form may also include vitamins or related nutrients, such as vitamin A, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, vitamin B12, carotenoids, niacin, folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, salts thereof, and derivatives thereof, and combinations thereof.

[0420] The nutritional formulations in powder or liquid form may also contain minerals such as phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium, chloride, and combinations thereof.

[0421] Nutritional formulations in powder or liquid form may also contain one or more masking agents to reduce the bitter taste, for example, in the reconstituted powder.

[0422] Suitable masking agents include natural and artificial sweeteners, sodium sources such as sodium chloride, and hydrocolloids such as guar gum, xanthan gum, carrageenan, and combinations thereof.

[0423] The amount of masking agent in the powdered nutritional formulation may vary depending on the specific masking agent selected, the other ingredients of the formulation, and other formulation variables or target product.

[0424] Process for producing a nutritional formulation in powder form according to the invention

[0425] The basic nutritional powder (comprising the pea protein isolate according to the present invention) can be prepared by dry blending all ingredients which are themselves in powder form.

[0426] As a variation, the base nutritional powder may be prepared using a conventional wet process which typically involves the use of two or more suspensions which are ultimately mixed, processed and then dried.

[0427] At least some of the plant protein present in the nutritional formulation in dry blend powder form is pea protein isolate, which has advantageously been dried by atomization prior to dry blending with a nutritional base powder, which typically comprises at least carbohydrates, vitamins, and minerals.

[0428] In certain embodiments, the pea protein isolate may be processed at high temperature for a short time (HTST) and then pasteurized prior to drying by atomization.

[0429] More specifically, the pea protein isolate can be added to water and hydrated; the water can be heated or unheated.

[0430] The suspension is then processed by HTST followed by drying by atomization. Optionally, the pea protein isolate can be conventionally homogenized after HTST treatment and before drying by atomization. For example, the isolate can be pasteurized at a temperature between 130° C. and 150° C. for a period of about 1 second to about 30 seconds.

[0431] The step of drying by spraying is a conventional step of drying by spraying, which is carried out at well known and conventional temperatures and times to produce spray-dried vegetable proteins.

[0432] Fields of use of nutritional preparations in powder form

[0433] The nutritional formulation in dry-blended powder form comprising the pea protein isolate according to the present invention has an improved mouthfeel after rehydration.

[0434] An individual may preferably consume at least one portion of the rehydrated nutritional formulation in powder form daily, and in certain embodiments, may consume two, three, or even more portions daily.

[0435] Each portion is preferably administered as a single dose, although the portion may also be divided into two or more localized portions and taken two or more times during the course of the day.

[0436] Nutritional formulations in powder form can be reconstituted for use by infants, children, and adults.

[0437] The term "about" refers to a value plus or minus 10%, preferably plus or minus 5%. BRIEF DESCRIPTION OF THE DRAWINGS

[0438] Figure 1 : Particle size distribution of the emulsion forming the nutritional formulation for clinical nutrition according to the present invention

[0439] Figure 2 : Sensory analysis of powder mixtures formulated with pea protein isolate according to the present invention

[0440] Figure 3 : Sensory analysis of ready-to-drink beverages for clinical nutrition

[0441] Figure 4 : Monitoring viscosity during in vitro digestion of pea protein isolate according to the present invention

[0442] Figure 5 :The solubility curve of pea protein isolate changes with pH

[0443] Figure 6 : Sensory analysis of ready-to-drink sports beverages

[0444] Figure 7 : Sensory analysis of dessert cream for clinical nutrition

[0445] Figure 8 : Fat globule size distribution of emulsions prepared with 100% milk protein for ice dessert preparation

[0446] Figure 9 : 50% milk protein and 50% pea protein for ice dessert preparation Size distribution of fat globules in emulsions prepared with S85F

[0447] Figure 10 : Size distribution of fat globules in an emulsion prepared with 50% milk protein and 50% pea protein isolate No. 1 according to the present invention for ice dessert preparation

[0448] Figure 11 : Size distribution of fat globules in an emulsion prepared with 50% milk protein and 50% pea protein isolate No. 2 according to the present invention for ice dessert preparation

[0449] Figure 12 : Melting curve of vegetarian ice cream prepared with pea protein isolate according to the present invention

[0450] Figure 13 : Sensory analysis of ice desserts

[0451] Figure 14 : Solubility of pea protein isolate compared to sodium caseinate changes with pH

[0452] Figure 15 : Sensory Analysis of Stirred Yogurt - Taste

[0453] Figure 16 : Sensory Analysis of Stirred Yogurt - Texture

[0454] Figure 17 : Sensory analysis of strawberry-flavored dairy beverages: Taste

[0455] Figure 18 : Sensory analysis of strawberry-flavored dairy beverages: Texture

[0456] Figure 19 :Viscosity analysis of chocolate muffin dough

[0457] Figure 20: Viscosity analysis of pancake batter

[0458] Figure 21 : Digestibility analysis by monitoring viscosity using the SIMPHYD device from NIZO

[0459] Figure 22 : sensory descriptors

[0460] The present invention will be more clearly understood with the aid of the following examples, which are intended to be illustrative rather than limiting.

[0461] Examples

[0462] Materials and Methods

[0463] Measurement of DH (degree of hydrolysis)

[0464] The measurement is based on the method for determining amino nitrogen on proteins and protein isolates according to the invention and calculating the degree of hydrolysis using the MEGAZYME kit (reference K-PANOPA).

[0465] principle:

[0466] The "amino nitrogen" groups of the free amino acids of the sample react with N-acetyl-L-cysteine and o-phthalaldehyde (OPA) to form isoindole derivatives.

[0467] The amount of the isoindole derivative formed in the reaction is stoichiometric with respect to the amount of free amino nitrogen.The isoindole derivative is measured by the increase in absorbance at 340 nm.

[0468] program:

[0469] An accurately weighed test sample P* of the sample to be analyzed is introduced into a 100 ml beaker. (This test sample is from 0.5 g to 5.0 g, depending on the amino nitrogen content of the sample.)

[0470] Add about 50 ml of distilled water, homogenize and transfer to a 100 ml graduated cylinder, add 5 ml of 20% SDS and make up the volume with distilled water; stir on a magnetic stirrer at 1000 rpm for 15 minutes.

[0471] Dissolve one tablet from bottle 1 of the Megazyme kit in 3 ml of distilled water and stir until completely dissolved. Provide one tablet per test.

[0472] This solution No. 1 was prepared extemporaneously.

[0473] Reactions are performed directly in the spectrophotometer cuvette.

[0474] ○Blank:

[0475] Introduce 3.00 ml of solution No. 1 and 50 μl of distilled water.

[0476] ○Standard material:

[0477] Introduce 3.00 ml of solution No. 1 and 50 μl of bottle No. 3 of the Megazyme kit.

[0478] ○Sample:

[0479] 3.00 ml of solution No. 1 and 50 μl of sample preparation were introduced.

[0480] The cuvette was mixed and the absorbance measurement of the solution was read after approximately 2 minutes at 340 nm on a spectrophotometer (the spectrophotometer was equipped with a cuvette with a 1.0 cm optical path, which can measure at a wavelength of 340 nm and was validated according to the procedure described in the manufacturer's technical manual associated therewith).

[0481] The reaction was immediately started by adding 100 μl of OPA solution bottle 2 from the Megazyme kit to the spectrophotometer cuvette.

[0482] The cuvette was mixed and placed in the dark for approximately 20 minutes.

[0483] Next, read absorbance measurements of the blank, standards, and samples at 340 nm on a spectrophotometer.

[0484] Calculation method:

[0485] The free amino nitrogen content, expressed as a percentage by mass of the product itself, is given by the following formula:

[0486]

[0487] Where: ΔA=A2-A1

[0488] V = volume of the bottle

[0489] m = mass of the test sample in g

[0490] 6803 = Extinction coefficient of isoindole derivatives at 340 nm (in L.mol -1 .cm -1 as units).

[0491] 14.01 = molar mass of nitrogen (in g.mol -1 (unit)

[0492] 3.15 = Final volume in the cuvette (in ml)

[0493] 0.05 = Test sample in the cuvette (in ml)

[0494] The degree of hydrolysis (DH) is given by the formula:

[0495]

[0496] Protein nitrogen is determined according to ISO 16634 and the DUMAS method.

[0497] Measuring solubility in water at various pH values

[0498] The measurement is based on diluting the sample in distilled water, centrifuging it and analyzing the supernatant.

[0499] program:

[0500] 150 g of distilled water at a temperature of 20° C.±2° C. are introduced into a 400 ml beaker, mixed with a magnetic bar and precisely 5 g of the test sample are added.

[0501] If necessary, adjust the pH to the desired value with 0.1 N NaOH.

[0502] Make up the content to 200 g with water.

[0503] Mix at 1000 rpm for 30 minutes and centrifuge at 3000 g for 15 minutes.

[0504] 25 g of supernatant was collected.

[0505] A pre-dried and tared crystallizing dish was introduced.

[0506] Place in an oven at 103℃±2℃ for 1 hour.

[0507] Then, the sample was placed in a desiccator (containing a dehydrating agent) to cool to room temperature, and then weighed.

[0508] The soluble solids content expressed as a weight percent is given by the formula:

[0509] (m1-m2)x(200+P)x 100

[0510] ………………………………=% solubility

[0511] P1 x P

[0512] in:

[0513] o P = weight of sample in g = 5 g

[0514] o m1 = weight of the crystallizing dish after drying, in g

[0515] o m2 = weight of the empty crystallizing dish in grams

[0516] o P1 = weight of sample collected in g = 25 g

[0517] Measuring in vitro digestibility

[0518] The SIMPHYD device from NIZO is a static model that simulates digestive processes in the gastrointestinal tract.

[0519] Gastric digestion is combined with online viscosity measurement over time. Adapted to physiological conditions, gastric acidification is initiated with concentrated HCl and enzymatic digestion enzymes (pepsin and lipase) are added.

[0520] All samples were subjected to the SIMPHYD apparatus at a concentration of 3% (m / v).

[0521] The measurements are as follows:

[0522] o Viscosity baseline determined over 5 minutes at natural pH and 37°C

[0523] o Then it was acidified to pH 2 with HCl and the system was kept at 37 °C for 15 minutes.

[0524] o Add pepsin and lipase at 20 minutes.

[0525] The rheometer was used in AR-2000TA instrument at 75 s -1 The viscosity was monitored at a shear rate of 100 for 3 hours.

[0526] The measurements were performed in duplicate. If the difference between the two measurements was too great, a third measurement was performed.

[0527] The profiles of the test proteins were compared with those established by Hall et al. (2003, Casein and whey exert different effects on plasma amino acid profiles, gastrointestinal hormone secretion and appetite, Br. J. Nutr. 89:239-248) for "fast" and "slow" proteins (whey protein and sodium caseinate, respectively).

[0528] The viscosity curve obtained is shown in Figure 21 middle.

[0529] The apparent viscosity of the control whey protein sample did not change during gastric administration, whereas the apparent viscosity of the sodium caseinate control increased after gastric acidification and remained high after the addition of digestive enzymes.

[0530] After 5 minutes of acidification, pea protein ( S85M) shows a first viscosity peak, then a second viscosity peak at 15 minutes, and then the viscosity curve rejoins with that of whey protein at slightly higher values.

[0531] The viscosity begins to decrease before the digestive enzymes are added.

[0532] The pea protein isolate according to the invention showed a very small increase in apparent viscosity which again decreased to a value slightly above that of whey protein over the course of 30 minutes.

[0533] The behavior of pea protein isolates according to the present invention reflects their characteristic "fast" nature as more satiating proteins than "slow" proteins. This results in faster gastric emptying and a post-absorptive increase in plasma amino acids.

[0534] Emulsifying ability measurement

[0535] The resulting emulsions were analyzed for the size of the fat globules formed using a particle size analyzer, as described above, as measured by light scattering of the reconstituted protein powder.

[0536] The results are expressed as:

[0537] ○Dmode, the diameter of the main group,

[0538] ○D(4.3), arithmetic mean diameter

[0539] o D10, D50 and D90, the diameters with 10%, 50% and 90% passing.

[0540] The following table summarizes the sizes of fat globules in emulsions prepared using the following methods:

[0541] ○ Two pea protein isolates according to the present invention, No. 1 and No. 2,

[0542] ○Various milk proteins

[0543] ○A batch of sodium caseinate.

[0544] The ΔD corresponds to the difference between D90 and D10; it reflects the dispersion state of the emulsion.

[0545] The smaller the value, the closer the droplet sizes are and the more uniform the emulsion.

[0546]

[0547] The pea protein isolate according to the present invention has:

[0548] ○Good emulsification properties (lower Dmode: 23.4 and 23.6 μm respectively)

[0549] ○ Emulsion stability (ΔD) on par with or even lower than some milk protein concentrates or sodium caseinate and

[0550] ○ Emulsion homogeneity is equivalent to the homogeneity of milk protein.

[0551] Furthermore, their properties make them quite transferable to applications where a certain level of emulsifying power is required, such as iced dessert preparations or non-dairy coffee washes, where caseinates are sought.

[0552] Example 1: Preparation of pea protein isolate according to the present invention and reference according to the present invention “ 1 ” and “ 2 ” of Characterization of pea protein isolate

[0553] Process for preparing pea protein isolate No. 1 according to the present invention

[0554] 1500kg of pea protein (produced by the applicant company under the trade name S85F) was mixed into 8500 litres of water preheated to 55°C.

[0555] The mixture was stirred at 55°C for 3 hours.

[0556] 0.5% (w / w) of the endoprotease FLAVORPRO 750 MDP (from BIOCATALYST) was added.

[0557] The mixture was stirred at 55°C for 1 hour.

[0558] A degree of hydrolysis of 7 was then obtained.

[0559] The reaction was quenched by heating the medium to 70°C and keeping it at this temperature for at least 10 minutes.

[0560] A UHT treatment was applied (protocol: 140°C - 10 seconds).

[0561] The mixture was dried by atomization to a solids content of approximately 93%.

[0562] Process for preparing pea protein isolate No. 2 according to the present invention

[0563] 1500kg of pea protein (produced by the applicant company under the trade name S85F) was mixed into 8500 litres of water preheated to 55°C.

[0564] The mixture was stirred at 55°C for 3 hours.

[0565] 0.3% (w / w) of endoprotease ENZECO FUNGAL PROTEASE (from EDC) was added.

[0566] The mixture was stirred at 55° C. for 1 hour, and the degree of hydrolysis then obtained was 6.5.

[0567] The enzyme reaction was inhibited by heating the medium to 70°C and keeping it at this temperature for at least 10 minutes.

[0568] A UHT treatment was applied (protocol: 140°C - 10 seconds).

[0569] The mixture was then dried by atomization to a solids content of approximately 93%.

[0570] Characteristics of the pea protein isolate prepared therefrom

[0571] 1. Free amino acid content

[0572] Measured according to standard NF EN ISO13903:2005

[0573]

[0574] 2. Viscosity curve

[0575] To determine the viscosity curve in water, the following measurements are required

[0576] o In an aqueous solution containing pea protein isolate at 15% solids (permeated and azide-treated water at 200 ppm to prevent any bacteriological risk),

[0577] o Using an AR2000 rheometer from TA Instruments,

[0578] oIt has a concentric cylindrical geometry,

[0579] o 600s in 3 minutes -1 0.6×10 -3 shear rate (log), and

[0580] o Temperature of 20°C (temperature equilibration for 3 minutes before testing).

[0581] Prior to measurement, the solution was stirred at 750 rpm and 20° C. for at least 10 hours.

[0582] No pH adjustment was performed.

[0583] The following table compares pea protein isolate according to the present invention with control milk protein and pea protein Viscosity curve of S85F.

[0584]

[0585]

[0586] It was found that the pea protein isolate according to the present invention showed Newtonian behavior like milk protein, while the pea protein S85F showed very obvious shear thinning behavior.

[0587] In addition, the viscosity of pea protein isolates No. 1 and No. 2 is very close to that of milk protein, or even lower.

[0588] 3. Changes of solubility curve in water with pH

[0589] The results are shown in the table below and are given by Figure 5 illustrate.

[0590]

[0591] 4. Stability study

[0592] A study of the stability of pea protein isolates according to the invention over time was performed to measure their behavior towards intact pea protein.

[0593] The study was conducted after six months of storage according to the following temperature / relative humidity regime:

[0594] ○40℃±2℃

[0595] ○At 75%±5% relative humidity.

[0596] The measurements are expressed as percent loss of solubility (measured according to the procedure described above).

[0597]

[0598] It was found that at pH 7, S85F lost about half of its solubility, while pea protein isolate lost at most one-fifth of its solubility and in all cases maintained a 10% higher solubility than the initial S85F has higher solubility.

[0599] 4-Digestibility curve

[0600] The purpose of this study was to evaluate the total protein digestibility of pea protein isolates No. 1 and No. 2 according to the present invention and to compare it with S85F for comparison.

[0601] For this study, 48 Sprague Dawley rats (Charles River, Lyon, France) weighing 100-125 g at the beginning of the study were randomly divided into four groups of 12 rats each according to their body weight.

[0602] The experiments were performed in accordance with the European Animal Experimentation Code and with reference to the Animal Health Act (APAFIS project number 0000501).

[0603] Upon their arrival, the rats underwent a 7-day quarantine period during which they received a standard chow diet for growing rats.

[0604] Starting on the first day of the study, the rats received the following diet for 10 days:

[0605]

[0606]

[0607] Amounts are expressed as weight percent.

[0608] Feed and drink consumption and body weight changes were monitored on the first and fifth days of the study and then daily until the tenth and final day of the study.

[0609] Urine and feces were also collected daily during the first five days of the study. The protein content of feed and feces was determined by Kjeldahl method (standard ISO 1871:2009).

[0610] Nitrogen analysis of feces and feed makes it possible to calculate the coefficient of digestibility and utilization (CDU):

[0611]

[0612] All rats showed the expected growth, which was significantly reduced in the protein-deficient control group, as always in this experimental protocol.

[0613] Beverage consumption was not altered by the various diets.

[0614] Changes in other urine and fecal parameters were directly related to the control or experimental diet.

[0615] For each experimental day, the following digestibility rates were calculated:

[0616]

[0617] From a statistical point of view, The protein digestibility of S85F was significantly different from that of pea protein isolate No. 1 according to the present invention (p=0.0003).

[0618] However, from a biological perspective, these differences are completely insignificant.

[0619] It can therefore be concluded that the digestibility between NUTRALYS and Pea Protein Isolates No. 1 and No. 2 according to the invention is similar using the following rounding:

[0620]

[0621]

[0622] 5-Digestive dynamics

[0623] This test uses an in vitro technique that simulates protein digestion according to the following method.

[0624] The use of in vitro digestion methods allows for the efficient screening of various protein-rich foods based on their physicochemical properties and their behavior during transit through the stomach and small intestine.

[0625] Here, compared A 3% (m / m) protein solution of S85F, pea protein isolates No. 1 and No. 2 according to the present invention and controls commonly used in such tests, namely casein and whey.

[0626] Therefore, these five solutions were tested in an in vitro model of dynamic digestion under physiological conditions equivalent to those of the stomach and then the small intestine.

[0627] The digestion model was coupled with real-time viscosity monitoring using a controlled stress rheometer (AR-2000, TA Instruments, New Castle, TX, USA) equipped with a stainless steel finned rotor (39 mm height and 28 mm diameter).

[0628] The protein solution was tested under the same conditions, i.e., at 37 °C for 150 s. -1 Conventional shearing was performed for 3 hours.

[0629] Monitor the base viscosity for 5 minutes and then gradually acidify the solution to a pH between 1.5 and 2.

[0630] This acidification typically takes 15 minutes.

[0631] Once the pH of the solution stabilized between 1.5 and 2, an enzyme mixture of pepsin (Sigma-Aldrich, St. Louis, MO, USA) and lipase (Novozyme, Gladesaxe, Denmark) was added. Figure 4 shown.

[0632] Monitoring viscosity during in vitro digestion clearly reflects the digestion kinetics of proteins. Thus, digestion of whey does not cause changes in viscosity, as it is a rapidly digested protein. Casein, on the other hand, shows a significant increase in viscosity after acidification, reflecting a slow digestion.

[0633] A type of pea protein exhibits behavior between these two standards; it is called "fast intermediate."

[0634] However, pea protein isolates No. 1 and No. 2 according to the present invention again showed Intermediate behavior between S85F and whey.

[0635] It should be noted that the combination of fast-acting proteins and intermediate proteins can promote digestion and prolong the time for amino acids to diffuse in the blood circulation, which is beneficial to protein synthesis in muscles after prolonged exertion.

[0636] Example 2: Replacing milk protein with pea protein isolate in UHT-treated ready-to-drink beverages for clinical nutrition

[0637] Nutritional formulations based on milk protein, pea protein and pea protein isolate are shown in the table below:

[0638]

[0639]

[0640] Amounts are expressed as weight percent.

[0641] Its nutritional value per 100ml is as follows.

[0642]

[0643] Mineral composition (per 100ml):

[0644]

[0645]

[0646] The process used to make the beverage is as follows:

[0647] ○ Dry blend all powders (milk protein, pea protein, pea protein isolate, maltodextrin and sucrose),

[0648] ○ Weigh out 90% of water at 55℃,

[0649] ○ The powder mixture was added to water at 55°C, dispersed with a stirrer for 1 minute, and then mixed with a SILVERSON blender at 55°C for 30 minutes,

[0650] In a separate mixing container, dissolve the minerals with the remaining water at 50°C and add immediately to the solution.

[0651] o Add vanilla flavoring to the solution,

[0652] ○ Place lecithin and oil in a separate mixing container, stir and heat to about 55°C,

[0653] o After 35 minutes of hydration, add the lecithin and oil mixture to the main batch using shear at 10,000 rpm for 5 minutes,

[0654] o Preheat the product to 75°C / 3 litre batches in a beaker in a water bath (temperature rise in about 10 minutes) and homogenize on a 10 litre / hour homogenizer at 200 bar (one stage) (about 20 minutes per batch),

[0655] ○ Cool the product to 30°C for 5 minutes to 45 minutes depending on the batch,

[0656] ○ At 30°C, adjust to pH 6.8-7 with 30% sodium hydroxide,

[0657] ○ Heat the product again in a water bath at 75°C (temperature rise for 10 minutes),

[0658] o Sterilize the product at 142°C for 5 seconds in a tubular heat exchanger and then cool to 25°C in the heat exchanger,

[0659] ○Put the product into the bottle,

[0660] ○Store at +4°C.

[0661] The following analyses were performed on the preparations:

[0662] 1. Particle size analysis of emulsions formed from the nutritional formulations according to the invention

[0663] The aim here was to analyse aspects of a nutritional formulation based on pea protein isolate against a control based on milk protein.

[0664] The analysis was performed using a Reference 2000 particle size analyzer from MALVERN. The results obtained are shown in Figure 1 and in the following table (the values in the table are the average of three measurements).

[0665] Dmode is the main particle diameter. d10, d50 and d90 are the particle diameter values representing 10%, 50% and 90% of the total particles, respectively.

[0666]

[0667] Four samples showed a bimodal particle distribution. The first peak (first particle cluster) centered at 0.3 μm dominated the first three formulations. For the nutritional formulations with pea protein, this population was rare.

[0668] The second peak of the bimodal distribution (the second particle population) depends on the sample:

[0669] o For formulations No. 1 and No. 2 according to the invention, there is a population centered around 3 μm, but for formulation No. 1, the volume of the population is larger;

[0670] ○ For the control formulation: the second peak is centered at 10 μm;

[0671] o The nutritional formula with pea protein shows its difference by having a second peak centered at 46 μm.

[0672] It follows that protein isolates No. 1 and 2 make it possible to obtain emulsion sizes close to those of beverages obtained with milk proteins.

[0673] Furthermore, Pea Protein Isolate No. 2 even provided a better emulsion size distribution (less bimodal distribution and better emulsion stability, represented by the difference between D90 and D10) compared to Pea Protein Isolate No. 1.

[0674] 2. Viscosity analysis

[0675] The aim here was to show the stability of nutritional formulations based on pea protein isolates against controls based on milk protein and also to demonstrate the technical advantages in choosing these isolates over pea protein.

[0676] The measurement parameters are as follows:

[0677] ○Rheometer: Physica MCR301

[0678] ○Tool: Tool: Concentric cylinder CC27

[0679] ○Temperature: 20℃ (5 minutes to reach equilibrium)

[0680] ○ Shearing: 0.05s in 6 minutes -1 to 1000s -1

[0681] The results of the viscosity measurements are shown in the following table (pea protein isolate No. 1 according to the invention was analyzed here):

[0682]

[0683] Heat treatment does not affect the three nutritional formulations in the same way:

[0684] ○ For the control formulation, heat treatment resulted in a decrease in viscosity;

[0685] o For Formulation No. 1, there is an increase in viscosity and increased shear thinning behavior, but this viscosity increase is still low and close to the value of the control formulation.

[0686] In fact, it was found that before heat treatment, the control formulation had the highest viscosity, followed by the pea protein based formulation.

[0687] On the other hand, after heat treatment, the control formulation had the lowest viscosity, followed by formulation No. 1 according to the invention.

[0688] In conclusion, the nutritional formulation No. 1 according to the invention and the control formulation based on milk proteins had similar rheological behavior in terms of viscosity and heat resistance.

[0689] Other viscosity measurements were carried out on the same beverage recipe, this time also with a nutritional recipe prepared with Pea Protein Isolate No. 2 and after storage of the beverage at +4°C for one month (particularly with a change in the cooking method, in this case after heat treatment, in-line UHT sterilization at 20 l / h (142°C - 5 s) with in-phase homogenization at 200 bar down to 75°C).

[0690]

[0691]

[0692] These results show that all formulations had increased viscosity after one month at 4° C. Compared to the pea protein formulations, formulations No. 1 and No. 2 had viscosities of the same order of magnitude as the control formulation after one month, with very little increase in viscosity.

[0693] Beverages containing pea protein isolates No. 1 and No. 2 were much more stable than beverages containing pea protein and approached the stability of beverages containing dairy protein.

[0694] Example 3: Replacing dairy protein with pea protein isolate in UHT-treated ready-to-drink beverages for sports nutrition

[0695] The nutritional preparation has the following composition:

[0696]

[0697]

[0698] Amounts are expressed as weight percent.

[0699] The nutritional value per 100ml is as follows

[0700]

[0701] The conditions for preparing the beverage were the same as those of Example 2.

[0702] The viscosity of the ready-to-drink sports drink was measured.

[0703] The aim here was also to show the stability of nutritional formulations based on pea protein isolates against controls based on milk protein and also to demonstrate the technical advantages in choosing these isolates over pea protein.

[0704] The measurement parameters are as follows:

[0705] o Rheometer: Physica MCR301

[0706] oTool: Tool: Concentric cylinder CC27

[0707] o Temperature: 20°C (5 minutes to reach equilibrium)

[0708] ○ Shearing: 0.05s in 6 minutes -1 to 1000s -1

[0709] The results of the viscosity measurements are shown in the following table:

[0710]

[0711] From this we can infer:

[0712] - Before UHT, formulations containing pea protein isolate have an intermediate viscosity between milk protein and pea protein,

[0713] - After UHT, formulations containing pea protein isolate lost viscosity, while pea protein gained viscosity; milk protein remained the most viscous.

[0714] Therefore, pea protein isolate is more stable to heat treatment than pea protein and is more suitable for UHT RTD sports drinks due to its low viscosity, which is a desired property for UHT RTD sports drinks.

[0715] Example 4: Using pea protein isolate as a substitute for UHT-treated liquid nutritional formulations for enteral clinical nutrition Milk protein

[0716] The nutritional formulation then has the following composition:

[0717]

[0718]

[0719] Amounts are expressed as weight percent.

[0720] The nutritional value per 100ml is as follows.

[0721]

[0722] The process used to make the beverage is as follows:

[0723] o Mix caseinate in 50°C water, add protein, and mix on a magnetic plate for 10 minutes,

[0724] o Add carbohydrates and minerals while mixing with a SILVERSON blender,

[0725] o Add oil while mixing with a SILVERSON blender for 5 minutes (10000 rpm),

[0726] o Homogenization in a NIRO SOAVI 2K high pressure homogenizer (2 stages) at 60°C, 250 bar,

[0727] o Adjust the pH to 6.9 with 50% citric acid solution,

[0728] o Sterilize in glass bottles at 120 °C for 15 minutes in an autoclave,

[0729] ○ Cool to room temperature.

[0730] Under these operating conditions, pea protein isolate can be advantageously used to replace milk protein.

[0731] Example 5: Comparison of four powder blends formulated with pea protein isolate according to the present invention for athletes Sensory properties of objects

[0732] The team consists of 13 people.

[0733] This panel is qualified to taste products formulated with pea protein. The panel is trained to examine their performance in the following areas:

[0734] Ability to differentiate products

[0735] Consensus, correct use of descriptive terms

[0736] Repeatability, the ability to test a product submitted twice

[0737] Specifically, the group was trained in the correct use of sensory descriptors for taste and texture, such as:

[0738]

[0739] This approach also allowed them to comment on other descriptors not anticipated in this list.

[0740] product

[0741] This nutritional formulation is a powder mix for athletes, which has the following composition:

[0742]

[0743] Amounts are expressed as weight percent.

[0744] Immediately before consuming, rehydrate them in water at room temperature.

[0745] Test conditions

[0746] - In the sensory analysis laboratory: - Separate tasting booths, white walls, calm environment (favorable for concentration)

[0747] - White light (to have exactly the same product visuals)

[0748] - In the morning or late afternoon (when sensory abilities are at their highest)

[0749] - Products are provided anonymously using a three-digit code (to prevent the code from influencing product evaluations)

[0750] - Products are presented in a randomized fashion (to prevent order and persistence effects)

[0751] test

[0752] The method used to compare the results was the Flash curve (JM Sieffermann, 2000 - Le profil Flash: Unoutil rapide et innovant d'évaluation sensorielle descriptive. [The Flash Profile: a rapid and innovative tool for descriptive sensory evaluation.] In: L'innovation: de l'idée au succès [Innovation: from the idea to success] - 12th AGORAL Meeting. pp. 335-340, March 22-23, 2000. Paris, France: Lavoisier, Tec&Doc.).

[0753] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[0754] Examples:

[0755] Sensory Descriptors: Crunchy, such as Figure 22 shown

[0756] Data processing:

[0757] A statistical method suitable for this type of data is the multifactor analysis of the product data rows (J. Pagès, 1994 - Multiple factor analysis (AFMULT package) (Multifactor analysis (AFMULT package)). In: Computational Statistics & Data Analysis (Computational Statistics & Data Analysis), Vol. 18, No. 1, August 1994, pp. 121-140).

[0758] To make the results clearer, several MFAs were performed; both overall and according to each criterion (appearance, smell, taste, texture). The provided graphs summarize all the results provided by this method.

[0759] The analysis was performed using R software (publicly available):

[0760] R version 2.14.1 (2011-12-22)

[0761] Copyright (C) 2011 The R Foundation for Statistical Computing

[0762] ISBN 3-900051-07-0

[0763] 20 Platform: i386-pc-mingw32 / i386 (32-bit)

[0764] This software is a working environment that requires loading modules containing computational functions, such as the FactoMineR version 1.19 package.

[0765] result

[0766] like Figure 2 The results are shown in graphical form.

[0767] Three groups are distinguished: Pea protein marketed by COSUCRA pea protein S85F, and two pea protein isolates according to Example 1 of the present invention.

[0768] The panelists confirmed little difference between the two pea protein isolates according to Example 1 of the present invention.

[0769] They had a less gritty, pea and paper / cardboard appearance than the control product, and they were more bitter and had more strawberry / banana notes (the flavoring used in this formulation).

[0770] and The mixture stands out in terms of texture, since its application creates a foam formation.

[0771] and The S85F blend, on its own, has a prominent sweetness.

[0772] Example 6: Comparison of sensory properties of ready-to-drink beverages for clinical nutrition

[0773] The team consists of 14 people.

[0774] The panel, as in Example 3, was qualified to taste products formulated with pea protein. The panel was trained to examine their performance in the following areas:

[0775] Ability to differentiate products

[0776] Consensus, correct use of descriptive terms

[0777] Repeatability, the ability to test a product submitted twice

[0778] Specifically, the group was trained in the correct use of sensory descriptors for taste and texture, such as:

[0779]

[0780]

[0781] This approach also allowed them to comment on other descriptors not anticipated in this list.

[0782] product

[0783] This product is a ready-to-drink beverage, and its formula is the formula of Example 2.

[0784] They were presented to the panelists at room temperature.

[0785] Test conditions

[0786] - In the sensory analysis laboratory: - Separate tasting booths, white walls, calm environment (favorable for concentration)

[0787] - White light (to have exactly the same product visuals)

[0788] - In the morning or late afternoon (when sensory abilities are at their highest)

[0789] - Products are provided anonymously using a three-digit code (to prevent the code from influencing product evaluations)

[0790] - Products are presented in a randomized fashion (to prevent order and persistence effects)

[0791] practise

[0792] The method used to compare products is the Flash curve (JM Sieffermann, 2000).

[0793] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[0794] Examples:

[0795] Sensory Descriptors: Crunchy, such as Figure 22 shown

[0796] Data processing:

[0797] The statistical method suitable for this type of data processing is the Multivariate Analysis (MFA) of product data (J. Pagès, 1994). To clarify the results, several MFAs were performed; both overall and for each criterion (appearance, aroma, taste, texture). The provided graphs summarize all the results provided by this method.

[0798] The analysis was performed using R software (publicly available):

[0799] R version 2.14.1 (2011-12-22)

[0800] Copyright (C) 2011 The R Foundation for Statistical Computing

[0801] ISBN 3-900051-07-0

[0802] 20 Platform: i386-pc-mingw32 / i386 (32-bit)

[0803] This software is a working environment that requires loading modules containing calculation functions, such as the FactoMineR version 1.19 package.

[0804] result

[0805] like Figure 3 The results are shown in graphical form.

[0806] S85F was presented twice to test the reproducibility of this panel: As can be seen from the figure, the two points are close in the first dimension (maximum) but not in the second; this second dimension is therefore considered to be composed of measurement noise. Therefore, there is no significant difference between the two pea protein isolates according to the invention, as they are close in the first dimension.

[0807] It can be seen that the two pea protein isolates according to the present invention are S85F has more vanilla / caramel aroma and saltiness, which proves that its flavor is more pea / vegetable and astringent; in texture it is more creamy / coated, thick and sticky.

[0808] Example 7: Comparison of sensory properties of ready-to-drink beverages for athletes

[0809] The team consists of 12 people.

[0810] The panel, as in Example 3, was qualified to taste products formulated with pea protein. The panel was trained to examine their performance in the following areas:

[0811] Ability to differentiate products

[0812] Consensus, correct use of descriptive terms

[0813] Repeatability, the ability to test a product submitted twice

[0814] product

[0815] The products were ready-to-drink beverages whose formulation was that of Example 3. They were presented to the panelists at room temperature.

[0816] practise

[0817] The method used to compare products is the Flash curve (JM Sieffermann, 2000).

[0818] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[0819] Examples:

[0820] Sensory descriptor: Fresh walnuts ,like Figure 22 shown

[0821] The following is a list of descriptors provided to group members as a guide:

[0822]

[0823]

[0824] Data processing:

[0825] A statistical method suitable for this type of data processing is the multivariate analysis of product records (J. Pagès, 1994). The set of descriptors generated by the evaluation body is a set of variables. The provided charts summarize all the results provided by this method.

[0826] Statistical analysis was performed using R software version 2.14.1 (2011-12-22).

[0827] result

[0828] like Figure 6 The results are shown in graphical form.

[0829] Two families were distinguished in dimension 1: two pea protein isolates / two pea proteins according to the invention, and for dimension 2, four samples could be characterized according to their texture, smell and taste.

[0830] Regarding texture, and The ready-to-drink beverage of S85F was thicker than the beverage with pea protein isolate according to the present invention.

[0831] Regarding the smell, the beverage with pea protein isolate No. 1 according to the present invention had more vanilla smell than the beverage with pea protein isolate No. 2, while for The smell is more of a pea smell.

[0832] About the taste, Seems spicy and chemical, and about the smell, more pea, walnut and vegetable smell. With PISANE and The ready-to-drink beverages of S85F have a common bitter taste and paper-cardboard nature.

[0833] Regarding the beverages with pea protein isolate according to the present invention, beverage No. 1 (with isolate No. 1) was more creamy and vanilla-scented, while beverage No. 2 was more cereal and milk jam / caramel-scented.

[0834] Example 8: Replacement of milk protein with pea protein isolate in UHT-treated dessert cream for clinical nutrition.

[0835] Nutritional formulations according to the invention based on milk, pea and competitive pea proteins and pea protein isolates are listed in the table below (in the order of 23% replacement):

[0836]

[0837]

[0838] Amounts are expressed as weight percent.

[0839] The nutritional value per 100g is as follows:

[0840]

[0841] The process used to make the beverage is as follows:

[0842] ○Preheat water to 50℃,

[0843] ○ Dry blend all powders (milk protein, pea protein, pea protein isolate, maltodextrin, dextrin, sucrose and starch),

[0844] ○ The powder mixture was added to water at 50°C, dispersed with a stirrer for 1 minute, and then mixed with a SILVERSON blender at 3000 rpm at 50°C for 30 minutes,

[0845] o adding flavoring to the solution,

[0846] o Place lecithin and oil in a separate mixing container, stir and heat to 50°C,

[0847] o After 30 minutes of hydration, the lecithin and oil mixture was added to the main batch using shear at 10,000 rpm for 5 minutes.

[0848] o The product is sterilized at 133°C for 55 seconds in a tubular heat exchanger and then packaged at 70°C,

[0849] oStore at 4°C.

[0850] Comparison of sensory properties of dessert creams for use in clinical nutrition.

[0851] The panel is qualified to taste the formulated products. The panel is trained to check their performance in terms of:

[0852] Ability to differentiate products

[0853] Consensus, correct use of descriptive terms

[0854] Repeatability, the ability to test a product submitted twice

[0855] The panel consisted of 26 people from the French company Roquette, and on the day of the tasting, a total of 11 people participated, six of whom had received special training in dessert cream.

[0856] The product was prepared and then stored in a refrigerator.

[0857] They were presented to the panelists at room temperature.

[0858] Test conditions

[0859] In the sensory analysis lab: individual tasting booths, white walls, a calm environment (favorable for concentration)

[0860] White light (to have exactly the same product visual effect)

[0861] In the morning or late afternoon (when sensory abilities are at their highest)

[0862] Products are provided anonymously using a three-digit code (to prevent the code from influencing product evaluations)

[0863] Products are presented in a randomized fashion (to prevent order and persistence effects)

[0864] practise

[0865] The method used to compare products is the Flash curve (JM Sieffermann, 2000).

[0866] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[0867] Examples:

[0868] Sensory Descriptors: Fangdeng sugar, such as Figure 22 shown

[0869] The following is a list of descriptors provided to group members as a guide:

[0870]

[0871]

[0872]

[0873] Data processing

[0874] The statistical method suitable for this type of data processing is the Multivariate Analysis (MFA) of product data (J. Pagès, 1994). To clarify the results, several MFAs were performed; both overall and for each criterion (appearance, aroma, taste, texture). The provided graphs summarize all the results provided by this method.

[0875] Statistical analysis was performed using R software version 2.14.1 (2011-12-22).

[0876] result

[0877] like Figure 7 As shown, the dessert cream was consistently distinguished by all panelists with a very high dimension 1 at almost 64%, which describes an extreme product in the following way.

[0878] The dairy control had the smoothest appearance and melted in the mouth, but was the thinnest and had the sweetest taste.

[0879] Regarding texture, use C9 and The dessert cream of S85F was thicker than the dessert cream using the pea protein isolate according to the present invention.

[0880] About taste, C9 and Compared to the S85F test, the dessert cream with pea protein isolate had less pea smell.

[0881] Example 9. Replacing dairy protein in ice cream / ice desserts with pea protein isolate

[0882] Four formulations were developed:

[0883] o Control: with 100% milk protein

[0884] o Recipe 1: Pea protein S85F replaces 50% of milk protein;

[0885] o Recipe No. 2: 50% of the milk protein was replaced with the pea protein isolate No. 1 according to the present invention; o Recipe No. 3: 50% of the milk protein was replaced with the pea protein isolate No. 2 according to the present invention;

[0886]

[0887] Amounts are expressed as weight percent.

[0888]

[0889]

[0890] The manufacturing process is as follows:

[0891] ○ Add skim milk to the container (40℃ / 45℃),

[0892] ○ Add the powder ingredients to the container and stir at 80 Hz for 15 minutes in a CHOCOTEC batch cooker.

[0893] ○ Mix the stabilizer and sugar together and combine the mixture into a container,

[0894] ○ Mix at 80 Hz for 20 minutes,

[0895] ○ Combine cream and glucose syrup,

[0896] ○ Mix at 80 Hz for 15 minutes,

[0897] ○Pasteurize at 80℃ for 3 minutes,

[0898] o Cool to 70°C - half of the resulting mixture was homogenized directly; the other half was cooled to 50°C. While the first batch was homogenizing, the second batch was heated to 70°C and then homogenized,

[0899] ○ Homogenization at 200 bar,

[0900] ○ Cool to 4°C in a aging container and add seasoning

[0901] Allow to mature for 23 hours

[0902] ○ Beat to obtain 95%-100% expansion and freeze at -30℃ for up to 1 hour

[0903] ○ Store ice cream at -20℃.

[0904] analyze

[0905] Characterizing mixtures during manufacturing

[0906]

[0907]

[0908] It should be noted that the swellability of the formulation prepared with the pea protein isolate according to the present invention was the same as that of the control and was not significantly different from the formulation prepared with pea protein.

[0909] Viscosity measurement

[0910]

[0911] The formulation with pea protein showed the highest viscosity. The formulation with pea protein isolate according to the invention was equivalent to the control formulation.

[0912] Particle size analysis

[0913] To evaluate the emulsifying capacity and stability of the emulsion, particle size analysis was performed at various steps in the preparation of ice cream:

[0914] o Size distribution of fat globules after the homogenization step,

[0915] o Size distribution of fat globules after the ripening step,

[0916] o Size distribution of ice cream fat globules (equivalent to the size distribution of fat globules after the beating step).

[0917] These analyses were also performed with the addition of 0.1% SDS to determine whether the emulsions were generated by aggregation / flocculation or coagulation.

[0918] The results are shown in Figures 8 to 11 middle.

[0919] For each formulation, the particle size distribution tended to decrease or become more unimodal after ripening.

[0920] For use of pea protein The change in the S85F formula is very obvious. This shows that pea protein S85F is the slowest emulsifier used to migrate on the surface of fat globules.

[0921] In contrast, recipe No. 3 (with pea protein isolate No. 2 according to the invention) was as good an emulsifier as the recipe comprising 100% milk protein.

[0922] Pea protein isolate No. 1 according to the invention has a poorer emulsification property than pea protein isolate No. 2 according to the invention after homogenization, but has a tendency to become equally good after ripening.

[0923] Example 10: Using pea protein isolate to completely replace dairy protein in ice cream / ice desserts

[0924] Three recipes were developed for these vegan ice creams:

[0925] ○ Control: 100% pea protein S85F,

[0926] Formula No. 1: 100% pea protein isolate No. 1 according to the present invention,

[0927] Formula No. 2: 100% pea protein isolate No. 2 according to the present invention

[0928]

[0929] Amounts are expressed as weight percent.

[0930] Nutritional value (per 100g) is as follows:

[0931]

[0932]

[0933] The manufacturing process is as follows:

[0934] ○Heat water to 45℃,

[0935] ○ Mixed ingredients,

[0936] ○ Mix the stabilizer with sucrose,

[0937] ○ Add water and mix for 20 minutes,

[0938] ○Introduce fat (melted coconut oil) and mixture,

[0939] ○Pasteurize at 80℃ for 3 minutes,

[0940] ○ Cool to 70℃,

[0941] o The mixture was homogenized at 200 bar (in two stages) - 30% in the second stage,

[0942] ○Add flavoring,

[0943] ○ Stir at 4℃ for 20 minutes to mature.

[0944] ○ Beat to 90%-100% and cool at -30℃ for 1 hour.

[0945] ○Store at -18°C.

[0946] analyze

[0947] Measurement of puffing capacity (ice desserts)

[0948] ○The weight of an empty crucible of a given volume V,

[0949] Measured mass = m c , where m c is the mass of the empty crucible

[0950] o The weight of a crucible of given volume V, filled to the brim with the mixture before puffing

[0951] Measured mass = mc + m 混合物 , where m 混合物 is the mass of the mixture corresponding to volume V

[0952] o The weight of a crucible of given volume V, filled to the brim with the mixture after puffing (taken from the refrigerator)

[0953] Measured mass = mc + m 冰 , where m 冰 is the mass of ice corresponding to volume V (puffed mix taken out of the freezer).

[0954] The bulking measurement is then given by:

[0955]

[0956] Characterization of the preparation process

[0957]

[0958] Viscosity measurement

[0959] oMeasured value at 4℃

[0960] o Rheometer: Physica MCR 301 Anton Paar

[0961] oGeometry: Concentric cylinder CC27

[0962] oNominal value: 0s within 5 minutes -1 to 200s -1

[0963]

[0964]

[0965] It is thus noted that the viscosity is lower when the formulation comprises pea protein isolate according to the invention.

[0966] Texture measurement

[0967] oMeasure temperature: When leaving the refrigerator,

[0968] o Rheometer: INSTRON 9506

[0969] oGeometry: Conical

[0970] oNominal value: Deformation applied for up to 20 minutes,

[0971]

[0972] It was found that the hardness was generally better for the formulations with the pea protein isolate according to the present invention. More specifically, the pea protein isolate No. 2 according to the present invention had a very high hardness, which was undoubtedly related to its higher puffing capacity (101%).

[0973] Measuring emulsions for mixes and ice desserts

[0974] plan:

[0975] o MALVERN 3000 Liquid Path Particle Size Analyzer (Particle Size Analyzer) (Solvent is demineralized water)

[0976] oOptical model: 1.46+0.001i, stirring speed is 1900 rpm.

[0977] The mixture before and after ripening was characterized with or without SDS:

[0978] o Without SDS: Introduce the sample directly into the particle size analyzer beaker containing only water,

[0979] o With SDS: 0.1% (i.e. 0.6 g) of SDS was introduced directly into the beaker of the particle size analyzer. After the SDS was dissolved, the sample was added for analysis.

[0980] The final ice cream is introduced into the bowl of the particle size analyzer without melting. After the ice cream has melted and dispersed, the measurement is performed.

[0981] The sizes of the emulsions before and after ripening with and without SDS are given in the table below.

[0982]

[0983] In the absence of SDS, the emulsion comprising the mixture of pea proteins (control) had a smaller particle size than the emulsion prepared from the pea protein isolate according to the present invention.

[0984]

[0985] Using SDS, fat aggregates are dispersed and therefore the Dmode three tests are closer. It should be noted that the formulation with Pea Protein Isolate No. 1 according to the invention has a particle size analysis peak with larger particles.

[0986]

[0987]

[0988]

[0989] No major changes were observed after ripening. The formulations with pea protein isolate according to the invention were more polydisperse than the formulations with pea proteins.

[0990] The emulsion size of the unmodified form of ice cream was measured in the absence of SDS.

[0991]

[0992] The main peak (Dmode) size of the three ice creams was similar. However, the formulations with pea protein isolate according to the invention were more polydisperse, especially the formulation with isolate No. 2.

[0993] Comparative studies were performed using commercial ice creams, which showed that these ice creams contained even higher amounts of coarse particles than the control recipe and recipes No. 1 and 2, which was related to their high content of fat globules.

[0994] Measuring melting behavior

[0995] plan:

[0996] Based on experience, a given volume of ice dessert sample is placed on the grid above the beaker. The following measurements are then taken:

[0997] o to the time when the first drop falls into the beaker,

[0998] o Percentage of ice cream melted over time (over 3 hours).

[0999] Figure 12 This clearly illustrates the fact that ice cream prepared with the pea protein isolate according to the invention melts less.

[1000] Sensory analysis

[1001] The team consists of 15 people.

[1002] This panel, as in the previous example, was qualified to taste products formulated with pea protein. The panel was trained to examine their performance in the following areas:

[1003] Ability to differentiate products

[1004] Consensus, correct use of descriptive terms

[1005] Repeatability, the ability to test a product submitted twice.

[1006] Compared to ice cream prepared with pea protein, the ice cream of the present invention has less bitterness, less pea smell, and less coloring.

[1007] The ice dessert with pea protein isolate No. 1 according to the present invention had some ice crystals and a more pronounced vanilla taste, was sweeter and had more fat than the other products.

[1008] The ice desserts with pea protein isolate No. 2 according to the present invention were sweet and fatty, and more creamy. They had a slightly more pronounced "green tea" taste.

[1009] in conclusion

[1010] During the production of ice desserts, the pea protein isolate according to the invention leads to lower viscosity compared to pea protein.

[1011] Isolate 1 had a harder texture, but this was not noticeable to the panelists.

[1012] Both isolates lead firstly to a reduction in the melting of the corresponding ice dessert.

[1013] In terms of taste, the best perception was given to the ice dessert prepared with Isolate No. 1, which had a sweet and distinct flavor, less bitterness, and less "pea" taste.

[1014] Example 11: Comparison of sensory properties of ice cream

[1015] The team consists of 20 people.

[1016] This panel is qualified to taste products formulated with pea protein. The panel is trained to examine their performance in the following areas:

[1017] Ability to differentiate products

[1018] Consensus, correct use of descriptive terms

[1019] Repeatability, the ability to test a product submitted twice

[1020] Specifically, the group was trained in the correct use of sensory descriptors for taste and texture, such as:

[1021]

[1022]

[1023] This approach also allowed them to comment on other descriptors not anticipated in this list.

[1024] The product ice creams were those of recipes No. 1, 2 and 3 of Example 9.

[1025] Test conditions

[1026] - In the sensory analysis laboratory: separate tasting booths, white walls, calm environment (favoring concentration) - white light (to have an identical product visual)

[1027] - In the morning or late afternoon (when sensory abilities are at their highest)

[1028] - Products are provided anonymously using a three-digit code (to prevent the code from influencing product evaluations)

[1029] - Products are presented in a randomized fashion (to prevent order and persistence effects)

[1030] practise

[1031] The method used to compare products is the Flash curve (JM Sieffermann, 2000).

[1032] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[1033] Examples:

[1034] Sensory Descriptors: Crunchy, such as Figure 22 shown

[1035] Data processing:

[1036] The statistical method suitable for this type of data processing is the Multivariate Analysis (MFA) of product data (J. Pagès, 1994). To clarify the results, several MFAs were performed; both overall and for each criterion (appearance, aroma, taste, texture). The provided graphs summarize all the results provided by this method.

[1037] The analysis was performed using R software (publicly available):

[1038] R version 2.14.1 (2011-12-22)

[1039] Copyright (C) 2011 The R Foundation for Statistical Computing

[1040] ISBN 3-900051-07-0

[1041] 20 Platform: i386-pc-mingw32 / i386 (32-bit)

[1042] This software is a working environment that requires loading modules containing calculation functions, such as the FactoMineR version 1.19 package.

[1043] result

[1044] The results are shown in Figure 13 middle.

[1045] All three samples were evaluated on the basis of creamy texture, coldness and fondant, and on the basis of pea, vanilla and bitterness.

[1046] However, some descriptors can distinguish them:

[1047] ·have The ice cream at S85F seemed harder and had a pea and cardboard taste.

[1048] • The ice cream with pea protein isolate No. 1 according to the invention was creamier and aerated with walnut flavor components.

[1049] • Ice cream with pea protein isolate No. 2 according to the invention was perceived as sweeter.

[1050] Example 12: Pea protein isolate “ Non-dairy coffee creamer / coffee water ” Use in matrix

[1051] a. 100% replacement for sodium caseinate

[1052] The aim here was to replace 100% of the sodium caseinate and obtain a product that was stable in coffee.

[1053] The measurement of emulsion viscosity after pasteurization and the measurement of coffee stability made it possible to show the relative Improvements in functional properties in its ability to replace sodium caseinate.

[1054] The following formulations were developed:

[1055]

[1056] Amounts are expressed as weight percent.

[1057] The manufacturing process is as follows:

[1058] o Melt the fat at 80°C while stirring constantly.

[1059] o Add Dimodan Hp to the melted fat to dissolve the monoglycerides,

[1060] o Heat 90% of the water to 50°C and add protein and hydrate while stirring continuously for 30 minutes.

[1061] o Dissolve phosphate in residual water at 40°C,

[1062] o After 30 minutes of hydration, add glucose syrup and phosphate to the main mix.

[1063] o Pre-emulsify the fat / Dimodan Hp mixture in the main mixture at 10000 rpm for 5 minutes,

[1064] o The product was placed in a Niro Panda 2K Soavi (GEA) high pressure homogenizer at 75°C, a first stage pressure of 160 bar and a second stage pressure of 30 bar,

[1065] o Pasteurize at 80°C for a few seconds and then place the product in cold water to stop the heat treatment.

[1066] The following analyses were performed on the preparations:

[1067] 1. Viscosity analysis

[1068] Viscosity measurements of the emulsion concentrates after the heat treatment step were performed at 65°C (normal atomization temperature).

[1069] equipment:

[1070] Physica MCR 301 Anton Paar rheometer

[1071] o Geometry: CC27

[1072] o Method 0s within 660s -1 to 1000s -1

[1073] The results obtained for various formulations are as follows:

[1074]

[1075] The viscosity of the emulsions of formulations 2 and 3 after pasteurization was closer to the milk control than that of formulation 4 prepared with pea protein, which made it possible to dry low viscosity emulsions with high solids content, here 60% by weight.

[1076] 2. Changes in solubility with pH

[1077]

[1078] Figure 14 The solubility variation with pH of pea protein isolate according to the present invention relative to caseinate is illustrated and reflects their superior behavior.

[1079] Assessing coffee stability

[1080] Rehydrating coffee

[1081] ○ Weigh out 2g of soluble coffee:

[1082] o Drinking water (calcium content 136 mg, and magnesium content 60 mg) was heated at 80° C., and 135 g of said water was added to the 2 g,

[1083] o Add 12.7g of concentrated emulsion to the coffee.

[1084] Flocculation in coffee appears to be less pronounced in the formulation containing the pea protein isolate according to the invention than in the formulation obtained with pea protein. However, this may be related to the improved solubility of the isolate relative to pea protein.

[1085] b. 50% replacement of sodium caseinate

[1086] The aim here was to replace 50% of the sodium caseinate and obtain a product that was stable in coffee.

[1087] The measurement of emulsion viscosity after pasteurization and the measurement of coffee stability made it possible to show the relative Improvements in functional properties in its ability to replace sodium caseinate.

[1088] The following formulations were developed:

[1089]

[1090]

[1091] The amounts are expressed as percentages by weight. The nutritional values per 100g are as follows.

[1092]

[1093]

[1094] The manufacturing process is as follows:

[1095] ○ Melt the fat at 80℃ while stirring constantly.

[1096] ○ Dissolve monoglycerides and diglycerides in liquid oil,

[1097] ○ Dissolve the powdered protein in water at 50°C within 30 minutes.

[1098] ○ Add glucose syrup and phosphates dissolved in part of the water,

[1099] ○ Pre-emulsify melted fat in aqueous solution by stirring at 10000 rpm,

[1100] ○Pasteurize at 80℃ for a few seconds,

[1101] o The product was placed in a Niro Panda 2K Soavi (GEA) high pressure homogenizer at 75°C, with a first stage pressure of 160 bar and a second stage pressure of 30 bar,

[1102] ○ Dilute the mixture to 50% solids to heat in the apparatus at 180°C (T 入口 ) and 90℃(T 出口) atomization, evaporation rate is 10l / h to 12l / h.

[1103] The following analyses were performed on the preparations:

[1104] 1) pH of the emulsion

[1105] pH Recipe No. 1 Recipe No. 2 Formula No. 3 25℃ on emulsion 7.69 9.24 8.72 75℃ on coffee 6.45 6.35 6.10

[1106] 2) Emulsion capacity

[1107] Measuring the size of the lipid globules (using a laser particle size analyzer) made it possible to determine the ability of the pea protein isolate according to the invention to form lipid globules of the smallest possible size.

[1108]

[1109] These results clearly show that the particle size distribution of the 50 / 50 blend is similar to that of the 100% caseinate control.

[1110] 3) Viscosity of an emulsion containing 60% solids at 65°C (before atomization)

[1111] equipment:

[1112] Physica MCR 301 Anton Paar rheometer

[1113] ○Geometric shape: CC27

[1114] ○ Method 0s within 660s -1 to 1000s -1

[1115]

[1116] The minimal viscosity of the 50 / 50 mixture makes it possible to atomize at higher solids levels than are normally required for caseinates.

[1117] 4) Stabilization of powdered “non-dairy coffee creamer” in coffee

[1118] Rehydrating coffee:

[1119] a. Weigh out 2g of soluble coffee

[1120] b. Add 8 g of emulsion and 150 ml of drinking water at 80°C (calcium content 136 mg, and magnesium content 60 mg).

[1121] The stability of the emulsion in coffee is determined by measuring the color change of the product - measuring color based on L (white balance), a (yellow balance) and b (green balance) coordinates. White color in coffee is one of the key standards sought by manufacturers and consumers.

[1122] The 2 point difference in the measured value of the L parameter for the coffee prepared with the 50 / 50 mixture (L=+96) over the control coffee prepared with caseinate (L=+98) reflects the superior stability of the mixture with the pea protein isolate according to the invention.

[1123] Example 13. Use of pea protein isolate in preparing stirred yogurt

[1124] The aim here is to replace 30% of the dairy protein.

[1125] The following formulations were developed:

[1126]

[1127] Amounts are expressed as weight percent.

[1128] solid 17.73 18.28 18.31 18.28 Total protein 3.70 3.70 3.70 3.70 milk protein 3.70 2.58 2.58 2.58 plant protein 0.00 1.12 1.12 1.12 lipids 1.01 1.01 1.01 1.01 carbohydrate 12.30 12.99 12.99 12.99 Of which sugar 10.98 10.14 10.14 10.14 Kcal / 100g 73.10 75.84 75.86 75.84 Degree of substitution 0.00 30.19 30.19 30.19

[1129] The manufacturing process is as follows:

[1130] o Heat water to 60℃,

[1131] o Add protein and allow to hydrate for 1 hour,

[1132] o Add the cream while mixing with a POLYTRON homogenizer for 2 minutes,

[1133] o Add the sugar / starch mixture over 10 to 15 minutes,

[1134] o High pressure homogenization at 75°C-80°C (two stages: 180 bar in the first stage - 200 bar in the second stage),

[1135] o Pasteurization was performed using a Power Point International tubular heat exchanger at 95°C, 6 minutes - 20 l / h,

[1136] o Add fermentation YF-L812-50U / 250L),

[1137] o Acidify to pH 4.6 at 42°C (acidification time 5-6 hours),

[1138] o Stir at 3600 rpm and 42 °C,

[1139] o Use Spindle 2G at 37℃ / 38℃, 3600rpm to smooth it out

[1140] o Place in a jar and store at 4°C.

[1141] Viscosity measurement

[1142]

[1143] Values are given within ±5%.

[1144]

[1145]

[1146]

[1147]

[1148] Formulation No. 3 has the closest behavior to the control formulation, however the viscosity curve is inverted relative to the viscosity change of the control formulation at D+7 and D+14.

[1149] Specifically, formulation No. 3 recovers viscosity at D+14 and is most shear-resistant at D+14.

[1150] Formulation No. 1 was more viscous and more shear-resistant than Formulation No. 3 at D+7, but this situation was reversed from D+14.

[1151] Formula No. 2, which contains pea protein isolate according to the present invention, was the most viscous of the four formulas and was more viscous than the control formula. Its viscosity decreased over time.

[1152] These results indicate that, due to its behaviour, the pea protein isolate according to the invention would make it possible to reduce the amount of starch in this formula if one wishes to make it similar to the viscosity of the control formula.

[1153] The same is true for formulations 1 and 3, but to a lesser extent.

[1154] Example 14: Comparison of sensory properties of stirred yogurt

[1155] For the taste evaluation, the panel consisted of 11 people, and for the texture evaluation, the panel consisted of 12 people.

[1156] These panels were qualified to taste products formulated with pea protein. They were trained to examine their performance in the following areas:

[1157] Ability to differentiate products

[1158] Consensus, correct use of descriptive terms

[1159] Repeatability, the ability to test a product submitted twice

[1160] Specifically, they were trained in the correct use of sensory descriptors for taste and texture, e.g.

[1161] Taste description:

[1162]

[1163] Texture descriptors

[1164]

[1165]

[1166] product

[1167] The three products tested in Example 11 (Control Formulation, Formulation 1, and Formulation 2) were evaluated three days after preparation and presented at a temperature of approximately 10°C (the products were stored in a refrigerator and evaluated upon removal).

[1168] Test conditions

[1169] - In the sensory analysis laboratory: - Separate tasting booths, white walls, calm environment (favorable for concentration)

[1170] - White light (to have exactly the same product visuals)

[1171] - In the morning or late afternoon (when sensory abilities are at their highest)

[1172] - Products are provided anonymously using a three-digit code (to prevent the code from influencing product evaluations)

[1173] - Products are presented in a randomized fashion (to prevent order and persistence effects)

[1174] practise

[1175] The method used to compare products is the Flash curve (JM Sieffermann, 2000).

[1176] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[1177] Examples:

[1178] Sensory Descriptors: Crunchy, such as Figure 22 shown

[1179] Two lists of descriptors related to taste or texture were proposed to the panellists as a guide: they are attached in the Appendix to this report.

[1180] Data processing:

[1181] The statistical method suitable for this type of data processing is the Multivariate Analysis (MFA) of product data (J. Pagès, 1994). To clarify the results, several MFAs were performed; both overall and for each criterion (appearance, aroma, taste, texture). The provided graphs summarize all the results provided by this method.

[1182] Statistical analysis was performed using R software version 2.14.1 (2011-12-22).

[1183] result:

[1184] The results are shown in Figure 15 (flavor) and Figure 16 (Texture) Medium:

[1185] ·Include The stirred yogurt at S85F had a runny and grainy texture in the mouth, with notes of peas, cardboard, and fresh walnuts;

[1186] Yogurt with dairy proteins appears fattier and creamier, thicker and grainier, and tastes more typically yogurty, sweeter and milky.

[1187] The yogurt with pea protein isolate No. 1 according to the present invention was between the control and the The S85F test items were among the best and stood out by having a taste of cereals and fermented milk as well as a special coating texture in the mouth.

[1188] Example 15. Use of pea protein isolate in preparing strawberry-flavored dairy beverages

[1189] The aim here is to replace 50% of the dairy protein.

[1190] The following formulations were developed:

[1191]

[1192]

[1193] The manufacturing process is as follows:

[1194] ○ Heat milk and water to 50℃,

[1195] ○ Add protein to the mixture,

[1196] ○Hydrolyze at 50°C with stirring for 1 hour,

[1197] ○After 45 minutes, preheat the fat to 50℃.

[1198] ○ Add sugar and starch to the main product,

[1199] ○Add coloring agents, flavoring agents and strawberry puree to the main products,

[1200] ○ Mix for 5 minutes,

[1201] ○ Pre-emulsify the fat in the main product at 10000rpm for 5 minutes,

[1202] ○ In-line homogenization of the product at 65°C-190 bar (two stages)

[1203] ○ Sterilize the product in a tubular heat exchanger at 30 liters / hour, 7 seconds residence time, 138°C

[1204] o Cool to 40°C and store at +4°C.

[1205] Example 16: Comparison of sensory properties of strawberry-flavored dairy beverages

[1206] For the taste evaluation, the panel consisted of 12 people.

[1207] These panels were qualified to taste products formulated with pea protein. They were trained to examine their performance in the following areas:

[1208] Ability to differentiate products

[1209] Consensus, correct use of descriptive terms

[1210] Repeatability, the ability to test a product submitted twice

[1211] Specifically, they were trained in the correct use of sensory descriptors for taste and texture, e.g.

[1212] List of description words:

[1213]

[1214]

[1215] Smell and taste

[1216]

[1217] Test conditions

[1218] - In the sensory analysis lab: individual tasting booths, white walls, calm environment (favorable for concentration)

[1219] - White light (to have exactly the same product visuals)

[1220] - In the morning or late afternoon (when sensory abilities are at their highest)

[1221] - Products are provided anonymously using a three-digit code (to prevent the code from influencing product evaluations)

[1222] - Products are presented in a randomized fashion (to prevent order and persistence effects)

[1223] practise

[1224] The method used to compare products is the Flash curve (JM Sieffermann, 2000).

[1225] The products are all presented at the same time. This is a question of comparing products by making a series of classifications: the panelists select the descriptors that are most relevant to them to distinguish the products and classify the products according to these descriptors; it is possible to group several products in the same row.

[1226] Examples:

[1227] Sensory Descriptors: Crunchy, such as Figure 22 shown

[1228] Two lists of descriptors related to taste or texture were proposed to the panellists as a guide: they are attached in the Appendix to this report.

[1229] Data processing:

[1230] The statistical method suitable for this type of data processing is the Multivariate Analysis (MFA) of product data (J. Pagès, 1994). To clarify the results, several MFAs were performed; both overall and for each criterion (appearance, aroma, taste, texture). The provided graphs summarize all the results provided by this method.

[1231] Statistical analysis was performed using R software version 2.14.1 (2011-12-22).

[1232] result:

[1233] The results are presented in Figure 17 (flavor) and Figure 18 (Appearance and texture in the mouth).

[1234] In terms of taste, the panelists clearly identified the control by identifying it as sweeter, creamier (smell and taste), and strawberry-like (smell and taste) than the test formulated with pea protein.

[1235] The test with the pea protein isolate No. 1 according to the invention was considered vegetable-cereal in terms of smell and taste, while maintaining a dairy smell, while the test with NUTRALYS retained a vegetable-pea smell and taste.

[1236] In terms of texture, all products are considered to be watery. Their characterization is essentially carried out on dimension 1; two groups are then distinguished:

[1237] - The milk control, which was judged to coat more in the mouth, was followed by The tests conducted,

[1238] - Pea protein isolate No. 1 according to the present invention was judged to be oilier.

[1239] Example 17: Protein-enriched biscuits (intended for professional / weight loss or sports nutrition)

[1240] The preparation has the following composition:

[1241]

[1242] Amounts are expressed as weight percent.

[1243] The nutritional values of these preparations are as follows:

[1244]

[1245] The manufacturing process is as follows:

[1246] - Dissolve the sodium bicarbonate and ammonium bicarbonate in the water. Add the sugar and glucose syrup and mix in a Hobart planetary mixer fitted with flat paddles at speed 1 for 1 minute to completely dissolve the sugar.

[1247] - Add fat and lecithin and mix at speed 2 for 2 minutes.

[1248] - Add the remaining powder all at once and mix at speed 1 for 2 minutes and then at speed 2 for 1 minute.

[1249] - Allow the dough to rest for 15 minutes to complete the hydration of the powder and homogenization of the product.

[1250] - The dough is placed into the hopper of the biscuit machine so that the dough is pressed in the mold cavity between two rollers to form biscuits.

[1251] - Recover the cookies on the conveyor belt and place them on the baking tray.

[1252] - Bake in a MIWE Econo fan oven at 170° C. (fan speed 2) for 9 minutes.

[1253] The analysis performed is as follows:

[1254] One of the first important criteria for producing cookies on a biscuit machine is the "workability" of the dough.

[1255] Over-hydrated dough will be sticky and will not release from the mold cavity.

[1256] Dough that is too dry will not fill the mold cavity and will form abnormal cookies.

[1257] Adding large amounts of protein will have an impact on dough texture. The table below illustrates the hydration adjustments needed to compensate for the incorporation of various proteins into cookie dough.

[1258] Specifically, proteins with a greater or lesser affinity for water will bind some of the water in the formulation. This water will then no longer be available to "plasticize" the dough, which will then be too dry to form. Therefore, increasing the hydration of the dough is essential to correct this deficiency.

[1259]

[1260] Unfortunately, in dry cookies (less than 3% moisture in the finished product), it is undesirable to add too much water because of the impact this will have on baking times and conditions.

[1261] Furthermore, adding more water will affect the sugar concentration and the kinetics of recrystallization. Now, this last point is a determining factor in cookie texture, especially crispness.

[1262] Therefore, soluble but low-functional proteins such as the pea protein isolate of the present invention make it possible to limit the correction to the addition of only +8% water instead of 12% for non-functional and insoluble proteins and more than 23% for soluble and functional proteins.

[1263] A quick sensory analysis of the produced biscuits gave the following results.

[1264]

[1265]

[1266] Example 18: Protein enrichment of chocolate chip muffins (intended for professional / weight loss or sports nutrition)

[1267] The preparation has the following composition:

[1268]

[1269] Amounts are expressed as weight in grams.

[1270] The manufacturing process is as follows:

[1271] - Heat mixture B to melt the chocolate

[1272] - Powder A was mixed in a Hobart planetary mixer equipped with flat paddles at speed 1 for 1 minute

[1273] - Add the melted Mix B to the powder and mix at speed 1 for 2 minutes

[1274] -Finally, add C and mix for 2 minutes at speed 1. Scrape the bowl and mix for another 2 minutes at speed 2

[1275] - Spread the product into paper muffin molds (70g per mold)

[1276] - Bake in a MIWE Econo fan oven at 180°C for 15 minutes (fan speed 2, air inlet closed).

[1277] The analysis performed is as follows:

[1278] Muffin batter viscosity:

[1279] The following characteristics were measured using a TA Instruments AR2000 rheometer:

[1280] Duration: 600s

[1281] Speed: 160 rpm

[1282] ○Temperature: 25℃.

[1283] The results are shown in Figure 19 middle.

[1284] In muffins, the viscosity of the product will have an impact on the rise during baking and thus the final volume. The pea protein isolate according to the present invention has a much lower viscosity than other pea proteins.

[1285] Example 19: Protein enrichment of instant pancake mix (intended for professional / weight loss or sports nutrition)

[1286] The aim here is to replace 50% of the dairy protein.

[1287] The preparation has the following composition:

[1288]

[1289]

[1290] Amounts are expressed as weight percent.

[1291] The nutritional values of these preparations are as follows:

[1292]

[1293] The manufacturing process is as follows:

[1294] Mix all powders

[1295] Add water and mix with a stirrer to obtain a homogeneous product

[1296] Allow to sit for 2 minutes

[1297] Cook in a griddle or multi-griddle cake skillet for about 2 minutes, flipping the pancakes when halfway done.

[1298] The analysis performed is as follows:

[1299] Impact on product viscosity

[1300] The following characteristics are measured using the RVA rheometer:

[1301] Time(s) Speed (rpm) Temperature (℃) 60 20 25 120 50 25 180 100 25

[1302] The results are shown in Figure 20 middle.

[1303] RVA viscosity measurements of protein-enriched preparations showed that those with pea protein isolate according to the present invention were less viscous than those with other pea proteins.

[1304] This effect on viscosity has an impact on the rising of the pancakes during cooking.

[1305] A quick sensory analysis of the produced pancakes gave the following results.

[1306]

[1307] Example 20: Protein-Rich Gluten-Free Bread

[1308] Traditional bread has a protein content of about 10%.

[1309] However, in gluten-free products, the protein content is very low. Protein supplementation of these products is then sought to rebalance the nutritional value through gluten-free proteins such as pea protein.

[1310] The preparation has the following composition:

[1311]

[1312]

[1313] The nutritional values of these preparations are as follows:

[1314]

[1315] The analysis performed is as follows:

[1316] Viscosity of dough obtained from four different sources of pea protein (including the isolate according to the invention):

[1317]

[1318]

[1319] The mixture corresponds to a gluten-free bread control. The results show that, with the exception of the pea protein isolate obtained according to the invention (which does not affect the viscosity of the product nor the final volume), the protein enrichment of this mixture has an effect on the viscosity and final volume (maximum height) of the product.

[1320] Example 21: Protein-enriched bread

[1321] The preparation has the following composition:

[1322]

[1323] Amounts are expressed as weight in grams.

[1324] The nutritional values of these preparations are as follows:

[1325]

[1326]

[1327] The analysis performed is as follows:

[1328] NUTRALYS Pea-BF Pea Protein Isolate No. 2 <![CDATA[Volume (cm 3 )]]> 1505 1745 Weight of bread after baking (g) 441.3 435.0 Moisture loss during baking 11.7％ 13.3％ Weight of three plates of breadcrumbs / 50mm diameter (g) 20.4 13.0 <![CDATA[Bread density (g / cm 3 )]]> 0.293 0.249 <![CDATA[Breadcrumb density (g / cm 3 )]]> 0.346 0.221

[1329] The volume and density are in favor of the pea protein isolate according to the invention, which allows for better rising and thus a more aerated and softer, less dense bread.

[1330] A rapid sensory analysis of the produced bread gave the following results.

[1331]

[1332] Example 22: High-protein potato chips (protein content > 60%)

[1333] High-protein potato chips are small grains obtained by extrusion, with a protein content greater than 60%.

[1334] These cereals are used as inclusions in cereal products such as cereal bars or clusters.

[1335] These high-protein chips are sometimes the only solution for protein enrichment of these cereal products, as incorporating protein in powder form has an undue impact on the texture of the finished product.

[1336] The technical difficulty of high-protein potato chips is to achieve a protein content greater than 60%, or even 70%, while maintaining crispness.

[1337] The crispness of extruded products is directly related to expansion. In cereals obtained by extrusion cooking, expansion occurs at the die exit under the pressure of water vapor.

[1338] A high protein crunchy formulation containing 75% protein has the following composition:

[1339]

[1340] Amounts are expressed as weight percent.

[1341] The procedure is as follows:

[1342] The potato chips or extruded cereals were obtained on a CLEXTRAL Evolum 25 brand co-rotating twin-screw extruder equipped with a shear screw profile.

[1343] For comparative testing, the parameters were set in the first stage to have only the “protein type” variable.

[1344] Extrusion was performed as follows:

[1345] Fixed parameters Screw speed (rpm) 300 Chopping speed (rpm) 800 Powder flow rate (kg / h) 6 Water flow rate (kg / h) 1.4 mold Three holes (3mm diameter), two of which are blocked Barrel temperature 1 to 6 (℃) 50-80-100-130-150-130 blade two

[1346] The analysis performed is as follows.

[1347] The method used to assess the quality of potato chips is based on the sum of the scores obtained regarding various appearance and texture criteria, according to the reference framework detailed below.

[1348] Get a first number about the overall appearance of the potato chip by summing:

[1349] - A number from 1 to 4 regarding the shape of the potato chip, where the number 1 represents a very irregular shape and the number 4 represents a very round shape.

[1350] - A number from 1 to 2 regarding the color, where 1 represents an unsatisfactory color (too dark / irregular) and 2 represents an acceptable color.

[1351] A second number, assessing texture and expansion level, is obtained by summing:

[1352] - A number from 1 to 5 regarding hardness, where the number 1 represents a "very hard" product / the number 3 represents a "crunchy" product and the number 5 represents a "brittle" product.

[1353] - A measure of the level of radial expansion established by the ratio of the average diameter of the potato chip to the die diameter.

[1354] The following table summarizes the results obtained. The highest scores indicate the best results.

[1355]

[1356] The above results show that the best product is obtained with the pea protein isolate obtained according to the present invention.

[1357] Microfunctional and slightly soluble proteins such as pea protein The product given by BF was average in terms of texture but unacceptable in terms of appearance. Soluble and functional proteins such as S85F gave mediocre results in terms of appearance and texture.

[1358] Example 23: High protein nutritional bar for athletes

[1359] The technical challenge of high protein nutritional bars is the challenge of controlling the texture during storage of the product.

[1360] The reason for this is that high protein bars have a tendency to harden over time.

[1361] Various hypotheses have been found in the literature to explain this phenomenon, in particular the migration of water between components and protein aggregation.

[1362] Therefore, the choice of protein or proteins is crucial to the quality of the finished product.

[1363] Prepare nutritional bars according to various recipes:

[1364]

[1365] Amounts are given as weight percent.

[1366] analyze:

[1367] Measuring the firmness of nutrition bars

[1368] Monitor the hardness after 1 month ( Determined on a needle penetration hardness tester - the force required for a knife to penetrate 40% of the thickness of the strip at a constant speed), the D values of the various formulations given above were measured. +1 、D +7 、D +14 、D +21 、D +28 .

[1369] Here are the results:

[1370]

[1371] Regardless of storage time, increasing levels of incorporation of Pea Protein Isolate No. 2 according to the present invention were inversely proportional to decreasing bar hardness.

[1372] For 37.5% of No. 2 pea protein isolate according to the present invention / 12.5% The best value was obtained for the ratio of S85XF / 50% WPC.

[1373] Example 24. Mozzarella-type vegetarian cheese containing pea protein isolate

[1374] Vegetarian cheese recipes containing pea protein isolate No. 2 according to the present invention are given in the table below.

[1375] The control was a formula containing NUTRALYS Pea Protein Type F85F.

[1376]

[1377]

[1378] Amounts are given as weight percent.

[1379] The process used to prepare the formulation is as follows:

[1380] o Add water to a container fitted with a heating jacket (such as a Stephan Bowl - www.stephan-machinery.com / index.php?id=3) and heat to 50°C,

[1381] o Add all powdered ingredients except citric acid,

[1382] o Mix at 750 rpm for 2 minutes at 50°C,

[1383] o Add oil and mix at 750 rpm for 2 minutes,

[1384] o Add citric acid and mix at 750 rpm for 1 minute,

[1385] o Heat the mixture to 75°C while mixing regularly by hand to prevent it from browning,

[1386] oPrevent steam from entering the sleeve,

[1387] o Cook for 5 minutes while mixing regularly,

[1388] oStop cooking and store at +6°C.

[1389] Analyses were conducted for colour, texture, “cutability” as well as freeze / thaw and melt stability.

[1390] While the color and texture of the two formulations were identical, the formulation with pea protein isolate No. 2 had better "shred" behavior and better melt stability. In addition, the flavor of formulation No. 2 was considered better.

Claims

1. A pea protein isolate: ○Contains 0.5%-2% free amino acids, ○ In water at 15% solids and 20°C, it has the following viscosities: ■In 10s -1 The shear rate ranged from 11×10 -3 Pa.s to 18×10 -3 Pa.s, In the 40s -1 The shear rate ranged from 9×10 -3 Pa.s to 16×10 -3 Pa.s, and ■In the 600s -1 The shear rate ranged from 8×10 -3 Pa.s to 16×10 -3 Pa.s, ○ Solubility in water at 20°C: ■ In the pH range from 4 to 5, the pH range is from 30% to 40%. ■40% to 70% in the pH range from 6 to 8; o Has a degree of hydrolysis (DH) of 5%-10%.

2. The pea protein isolate according to claim 1, having a digestibility expressed as a coefficient of digestibility and utilization (CDU) of 93.5% to 95%.

3. The pea protein isolate of claim 1 , which is characterized as a "fast viscosity" protein according to the SIMPHYD test, reflecting the rapid duodenal assimilation of the constituent amino acids of the isolate.

4. The pea protein isolate of claim 1 , which has been pasteurized at high temperature for a short time before being dried by atomization.

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