EP3945862A1 - Field bean protein composition - Google Patents

Abstract

The invention relates to the field of plant protein isolates, and in particular to field bean protein isolates. The invention also relates to a process for the production thereof and to industrial applications thereof.

Description

Description

Title: PROTEIN COMPOSITION OF FEVEROLE

Technical area

[1] The invention relates to the field of protein isolates from legumes, and in particular protein isolates from field beans.

Prior art

[2] Field beans, or field beans (according to the old spelling), are annual plants of the species Vicia faba. They are legumes from the Fabaceae family, a subfamily of the Faboideae, a tribe of the Fabeae.

[3] It is the same species as the bean, a plant used since antiquity for human consumption. The word bean therefore designates both the seed and the plant.

[4] Several production methods are known from the prior art which make it possible, starting from field bean seeds, to produce a protein isolate.

[5] “Potential of Fava Bean as future protein supply to partially replace meatus intake in the human diet.” (Multari & al., In Comprehensive Reviews in Food Science and Food Safety Vol. 14, 2015) provides an excellent review of current knowledge on this subject.

[6] The classic process is initiated by grinding the beans to obtain a flour. This is then diluted in water in order to undergo an alkaline extraction aimed at solubilizing the bean proteins. The solution then undergoes a liquid / solid separation in order to obtain on the one hand a crude protein solution and on the other hand a solid fraction enriched with starch and fibers. The proteins are extracted via isoelectric pH precipitation of the proteins, they are separated from the aqueous solution and dried.

[7] The protein isolate thus obtained has a protein content of at least 80% (expressed as total nitrogen multiplied by the coefficient 6.25, on the dry matter total, calculation method described in the document available at the following address: http: //www.favv- afsca.fgov.be/laboratories/methods/fasfc/_documents/METLFSAL003Proteinebrut ev10.pdf). This is of known long-term industrial interest, especially in human and animal food. Indeed, its nutritional and functional properties allow it to be included in a large number of recipes and formulations.

[8] However, there remain two major technical problems which one skilled in the art still has to face to this day.

[9] First of all, the protein isolate obtained is systematically characterized by a dark gray, even black color. This mainly comes from the tannins and polyphenols present in the external fibers, entrained with the proteins during the manufacturing process of said protein isolate.

[10] Despite extreme care, traditional methods of dehulling the outer fiber (known as "dehulling" in English) do not remove enough tannins and polyphenols, and the apparent dark coloration limits the number of possible applications.

[11] Optimized processes have been developed. The process described for example in “Technological-scale dehulling process to improve the nutritional value of faba beans” (Meijer & al., In Animal Feed Science and Technology, 46, 1994) involves two grindings, two filtrations and a turboseparation (classification of particles according to their density using an ascending air current). These technological refinements are complex and therefore expensive.

[12] The tannins and polyphenols being soluble at alkaline pH, one strategy also consists in not carrying out the alkaline extraction mentioned above. Unfortunately, if the solubilization of these compounds is thus limited and makes it possible to limit the dark coloration, the extraction yield is greatly limited. Indeed, the bean proteins being more soluble at alkaline pH, extraction at neutral or acidic pH limits the extraction yield. [13] Secondly, the faba bean protein isolate according to the prior art has a water retention of less than 3 grams per gram of protein. Water retention consists of the measurement of the quantity of water capable of being absorbed by the protein isolate after exposure of the latter to an aqueous solvent under conditions defined in Test A, described in detail in the pages following sections of this description.

[14] For example, in the article "Nutritional and functional properties of Vicia Faba protein isolates related fractions. "(Vioque, Food Chemistry, 132, 2012), the water retention capacity of the isolate is 2.55 grams per gram of protein (see Table 3 of the article). Likewise, in “Composition and functional properties of protein isolates obtained from commercial vegetables grown in northern Spain” (Fernandez-Quintela, in Plant Foods for Human Nutrition, 51, 1997), the water retention capacity of the isolate is of 1.8 grams per gram of protein (cf. Table 4 of the article).

[15] These values, suitable for certain industrial applications, may be limiting for others.

[16] It is therefore of interest for the technique to know a simple and effective process, allowing access to a bean isolate as clear as possible in coloring and having a water retention greater than 3 grams of water per. grams of isolate.

[17] It is to the Applicant’s credit for having found such a process and such an isolate. This invention will be described in the following section.

Description of the invention

[18] According to the present invention, a bean protein composition is proposed, the color of which is characterized by an L component greater than 70 according to the L * a * b measurement and the water retention is greater than 3 grams of water per gram of isolate.

[19] According to another aspect, there is provided a process for producing a bean protein composition according to the invention, characterized in that it comprises the following steps: 1) Use of bean seeds; 2) Grinding of the beans using a stone mill, followed by separation of the ground material obtained into two so-called light and heavy fractions using an ascending air flow, then a second grinding heavy fraction with a knife mill; 3) Final grinding of the heavy fraction using a roller mill to obtain a flour; 4) Suspending the flour in an aqueous solvent; 5) Removal of the solid fractions from the suspension by centrifugation and obtaining a liquid fraction; 6) Isolation by precipitation by heating at isoelectric pH of the bean proteins contained in the liquid fraction; 7) Dilution of the bean proteins previously obtained to 15-20% by weight of dry matter and neutralization of the pH between 6 and 8, preferably 7, to obtain the protein composition of bean; 8) Drying of the protein composition of faba beans.

[20] According to a final aspect, it is proposed the industrial uses, in particular in human or animal food, in cosmetics, in pharmacy, of the faba bean protein isolate according to the invention.

[21] The invention and the variants thereof may allow, in general, to provide a practical and efficient solution to meet the needs of the industry to have a protein isolate of faba bean whose color is characterized by an L component greater than 70 according to the L * a * b measurement and the water retention is greater than 3 grams of water per gram of isolate, of its production process and of suitable industrial uses.

[22] The invention will be better understood from the description, presented in the following chapters.

Brief description of the drawings

[23] Other features, details and advantages of the invention will become apparent on reading the detailed description below, and on analyzing the accompanying drawings, in which:

Fig. 1

[24] [Fig. 1] shows a conventional method of separating the outer fibers and cotyledons of bean seeds; Fig. 2

[25] [Fig. 2] shows a method according to the invention for separating the outer fibers and cotyledons of bean seeds;

Detailed description of the invention

[26] As mentioned above, according to the present invention, it is first of all proposed a bean protein composition whose color is characterized by an L component greater than 70, preferably greater than 75, even more preferably greater than 80, according to the L * a * b measurement and the water retention according to test A is greater than 3 grams, preferably greater than 3.5 grams of water per gram of isolate.

[27] By "faba bean" is meant the group of annual plants of the species Vicia faba, belonging to the group of legumes of the family Fabaceae, subfamily of Faboideae, tribe of Fabeae. We distinguish the Minor and Major varieties. In the present invention, wild varieties and those obtained by genetic engineering or variety selection are all excellent sources.

[28] By "protein composition" is meant any composition rich in proteins, obtained by extraction from a plant and purification if necessary. A distinction is made between concentrates whose richness expressed as% protein on dry matter is greater than 50%, and isolates whose richness expressed as% protein on dry matter is greater than 80%.

[29] By "L * a * b measurement" is meant the evaluation of the coloring according to the CIE (Commission Internationale de l'Eclairage) chromatic space methodology presented in the publication "Colorimetry" (n ° 15, 2nd Ed., P. 36, 1986), using an adapted spectrophotometer, which converts it into 3 parameters: the lightness L which takes values between 0 (black) and 100 (reference white); parameter a represents the value on a red green axis and parameter b represents the value on a blue yellow axis. The measurement of this coloration is preferably carried out using DATA COLOR -DATA FLASH 100 or KONIKA MINOLTA CM5 spectrophotometers, with the help of their user manuals. [30] By "water retention" is meant the amount of water in grams that a gram of protein is likely to absorb.

[31] To measure this water retention capacity, test A is used, the protocol of which is described below.

[32] Weigh 20 g of sample to be analyzed in a beaker, add drinking water at room temperature (20 ° C +/- 1 ° C) until the sample is completely submerged, leave in static contact for 30 minutes, separate the residual water and the sample using a sieve and weigh the final rehydrated weight P of the sample in grams.

[33] The following calculation is then applied to obtain the Water retention capacity (in g) = (P - 20) / 20

[34] Preferably, the isolate according to the invention is characterized by a high protein content greater than 70% expressed as a percentage by weight of proteins on dry matter, preferably greater than 80% by weight, even more preferably greater than 90 % in weight.

[35] Preferably, the protein composition according to the invention has a dry matter greater than 80% by weight, preferably greater than 85% by weight, even more preferably greater than 90% by weight. Any method for measuring the water content can be used to quantify this dry matter, the gravimetric technique evaluating the loss of water by desiccation is preferred.

It consists of determining the amount of water evaporated by heating a known quantity of a sample of known mass.

- We weigh the sample at the start and measure a mass m1 in g.

- The water is evaporated by placing the sample in a heated chamber until the mass of the sample stabilizes, the water being completely evaporated. Preferably, the temperature is 105 ° C under atmospheric pressure.

- We weigh the final sample and measure a mass m2 in g

- Dry matter = (m2 / m1) * 100. [36] A second aspect of the invention consists of a process for producing a faba bean protein composition according to the invention characterized in that it comprises the following steps: 1) Use of bean seeds; 2) Grinding of the beans using a stone mill, followed by separation of the ground material obtained into two so-called light and heavy fractions using an ascending air flow, then a second grinding of the heavy fraction with a knife mill; 3) Final grinding of the heavy fraction using a grinder selected from roller mills and knife mills to obtain a flour; 4) Suspending the flour in an aqueous solvent; 5) Removal of the solid fractions from the suspension by centrifugation and obtaining a liquid fraction; 6) Isolation by precipitation by heating at isoelectric pH of the bean proteins contained in the liquid fraction; 7) Dilution of the bean proteins previously obtained to 15-20% by weight of dry matter and neutralization of the pH between 6 and 8, preferably 7, to obtain the protein composition of bean; 8) Drying of the protein composition of faba beans.

[37] By "stone mill" is meant a system made up of two superimposed stone cylinders leaving a space approximately equal to the size of the seed. One of the cylinders is static, while the other is rotating. The seeds are introduced between these two cylinders, and their relative movement will impose a physical constraint on these seeds.

[38] The term "knife mill" should be understood to mean a system consisting of a chamber equipped with an upper inlet for introducing the seeds, several knives arranged on an axis intended to put them in rotation in said chamber and of a lower outlet equipped with a sieve to let out only the seeds of a desired grain size.

[39] The first step consists of the implementation of faba beans. These still include their protective external fibers, also called “hulls” in English. The seeds can undergo a pre-treatment which may include steps of cleaning, sieving (separation of the seeds from the stones, for example), soaking, bleaching or toasting. Of Preferably, if bleaching is performed, the heat treatment schedule will be 3 minutes at 80 ° C. Non-limiting examples of varieties are, for example, the Tiffany, FFS or YYY varieties. Preferably, varieties of bean seeds with a naturally low tannin and / or polyphenol content will be used, such as the Organdi variety. Such varieties are known and capable of being obtained by varietal crossing and / or genetic modifications.

[40] The second step aims at the most efficient possible separation of the external fibers and the cotyledons. It is initiated by the first crushing of the beans using a stone mill. A particular and particularly suitable example of such a stone mill is, for example, sold by the company Alma®. As previously described, the seed will be introduced into a space made up of two stone discs, one of which is rotating. The Applicant has noticed that this technique is of particular interest because it will cause a very efficient separation of the external fibers and the cotyledons of the seeds. Preferably, the inter-disc space is adjusted between 0.4 and 0.6 mm.

[41] The ground material obtained is then subjected to the application of an ascending air flow, against the current. The different solid particles will be classified according to their density. Typically, after equilibrium, two fractions are obtained: a light fraction mainly containing the external fibers or “hulls” and a “heavy” fraction mainly containing the cotyledons. A particular and particularly suitable example of a suitable device is, for example, MZMZ 1 - 40 sold by the company Hosokawa-alpine®.

[42] The heavy fraction, enriched in cotyledons, will then be ground using a knife mill. A particular and particularly suitable example of such a knife mill is, for example, the SM300 sold by the company Retsch®.

[43] The succession of the three operations mentioned above within the second step aims to separate very finely the external fibers and the cotyledons, avoiding degrading these two parts and mixing them. The processes of art prior art are either too simplistic and do not achieve effective separation of the external fibers, or complicated and therefore difficult to operate from an industrial point of view. The process described for example in “Technological-scale dehulling process to improve the nutritional value of faba beans” (Meijer & al., In Animal Feed Science and Technology, 46, 1994) involves two grindings, two filtrations and one turboseparation (by current rising air). This process makes it possible to obtain a cotyledon fraction which still contains 1.2% of external fibers in the cotyledons. Our invention simplifies the process (two grindings using types of grinders of different technologies, with a turboseparation between the two grindings) and allows to reduce the content of external fibers to a value of 1% or even less.

[44] The third step aims to reduce the particle size of the heavy fraction enriched in cotyledons by grinding them using a mill selected from among roller mills and knife mills, in particular a roller mill. A particular and particularly suitable example, for so-called "dry" grinding, that is to say without solvent, of such a roller mill is, for example, MLU 202, sold by the company Bühler®. It is used here in order to reduce the grain size of the flour overall, in order to obtain a homogeneous and sufficiently fine powder to allow the next step 4 to be facilitated. The preferred particle size is between 200 and 400 microns, preferably 300 microns. In order to measure this granulometry, a laser granulometry apparatus is preferably used, although any method is possible such as sieving.

[45] Alternatively, the step of reducing the particle size of the heavy fraction enriched in cotyledons, also called final grinding of the heavy fraction, can be carried out in the presence of an aqueous solvent, preferably water. In this case, the fourth step below is merged with the third step which are then carried out concomitantly. In this case, a suitable mill is for example the Hurschel® Comitrol 3000 knife mill.

[46] The fourth step aims to suspend the powder obtained in the third preceding step in an aqueous solvent, preferably in the water. The aim here is to achieve a selective extraction of certain compounds, mainly proteins as well as salts and sugars, by dissolving them. The pH of the solution is advantageously adjusted to an alkaline pH in order to maximize the solubilization of the proteins. This pH adjustment can be carried out before and / or after suspension of the powder in the aqueous solvent.

[47] The aqueous solvent is preferably water. The latter can nevertheless be additivated, for example with compounds making it possible to facilitate solubilization. The pH of the aqueous solvent is adjusted between 8 and 10, preferably 9. Any basic reagent such as sodium hydroxide or lime can be envisaged, but potassium hydroxide is preferred. The temperature is adjusted between 2 ° C and 30 ° C, preferably between 10 ° C and 30 ° C, preferentially between 15 ° C and 25 ° C, even more preferably at 20 ° C. This temperature is regulated throughout the extraction reaction.

[48] Alkaline pH is effective in maximizing protein solubilization. Unfortunately, tannins and / or polyphenols are also solubilized at alkaline pH. Certain bean extraction processes avoid this basic pH rectification, favoring limiting the yield to contamination by polyphenols. Our particular process carried out in the second step makes it possible to implement this alkaline extraction, without excessively dissolving the polyphenols.

[49] The powder obtained is diluted in order to obtain a suspension of between 5% and 25%, preferably between 5% and 15%, preferably between 7% and 13%, even more preferably between 9% and 11%, most preferred being 10%, the percentage being expressed in weight of powder per total weight of water / powder suspension. The suspension is stirred using any equipment well known to those skilled in the art, for example a tank equipped with an agitator, equipped with blades, marine propellers, or any equipment allowing efficient agitation. The extraction time, preferably with stirring, is between 5 and 25 minutes, preferably between 10 and 20 minutes, even more preferably 15 minutes. [50] The fifth step aims to separate by centrifugation the soluble fraction and the solid fraction obtained during the fourth step. The preferred industrial principle can be found in patent application EP 1400537 which is incorporated here by reference. The principle of this process is to start by using a hydrocyclone to extract a fraction enriched in starch, then to use a horizontal decanter in order to extract a fraction enriched in internal fibers. Nevertheless, it is possible to use an industrial centrifuge which will extract a fraction enriched with starch and internal fibers. In all cases, solid fractions are obtained and a liquid fraction which concentrates the majority of proteins.

[51] The sixth step aims to acidify the bean proteins to isoelectric pH, around 4.5, then to subject the solution to heating in order to coagulate the proteins called globulins, which will be separated by centrifugation.

[52] The acidification is carried out at a pH between 4 and 5, preferably 4.5. This is preferably carried out with hydrochloric acid at about 7% by mass, but all types of acids, mineral or organic, can be used, such as citric acid. Even more preferably, the use of pure ascorbic acid or in combination with another mineral or organic acid is also possible. The use of ascorbic acid to acidify allows an improvement of the final color. Any means of heating is then possible, for example by means of a stirred tank equipped with a jacket and / or coil or an in-line steam injection cooker ("jet cooker" in English). The heating temperature is advantageously between 45 ° C and 75 ° C, preferentially between 50 ° C and 70 ° C, even more preferably between 55 ° C and 65 ° C, the most preferred being 60 ° C. The heating time is advantageously between 5 minutes and 25 minutes, preferably between 10 and 20 minutes, the most preferred being 10 minutes.

[53] The protein composition, mainly globulin, will coagulate and precipitate within the solution. It will be separated by any centrifugation technique, such as the Flottwegg® Sedicanteur. The solution residual obtained concentrates sugars, salts and albumins, it is called bean soluble. It will be treated separately, preferably evaporated and / or dried.

[54] It should be noted that the prior art of protein extraction from faba bean teaches exclusively isoelectric precipitation, without heating. The combination of the two steps according to the invention makes it possible to obtain the isolate according to the invention, but also to obtain bean solubles (name of the supernatant obtained after precipitation and centrifugation) which are stable at temperature. Indeed, the bean solubles obtained by isoelectric precipitation when exposed to a high temperature, for example in an evaporator, will precipitate. This precipitate is a major disadvantage because it leads to clogging of industrial installations.

[55] In contrast, the combination of isoelectric precipitation with controlled heating proposed by the invention allows to obtain:

- a floc of coagulated proteins, giving after the required treatment the product claimed in this application, and

- residual solubles containing, among others, soluble proteins (albumins), salts and sugars

The second fraction can typically be upgraded in the fermentation and / or animal nutrition industries. To do this, it must be concentrated so that it is stabilized from a bacteriological point of view. To do this, a concentration operation by vacuum evaporation is conventional, carried out using a second heating separate from that which made it possible to coagulate the floc. During this operation, and in the event of simple isoelectric precipitation during the floc / soluble separation, a deposit of coagulated proteins will accumulate in the evaporator.

[56] In a seventh step, the protein composition is then diluted to approximately 15-20% by weight of dry matter and neutralized to a pH of between 6 and 8, preferably 7, using any type of basic agent, preferably potash at 20% by mass. [57] The protein composition can then undergo a heat treatment, preferably at a temperature of 135 ° C by direct injection of steam by nozzle and cooling by flash effect under vacuum at 65 ° C.

[58] The protein composition obtained can be used directly, for example by being hydrolyzed by a protease or else textured by an extruder.

[59] In an eighth step, the protein composition according to the invention is dried. The preferred mode of drying is atomization, particularly using a multi-effect atomizer. Typical settings are an inlet temperature of 200 ° C and a vapor temperature of 85-90 ° C.

[60] According to a final aspect, it is proposed the industrial uses, in particular in human or animal food, in cosmetics, in pharmacy, of the faba bean protein isolate according to the invention. The faba bean composition obtained according to the invention has a very high protein content as well as a very white coloring, thus making it possible to be included in a large number of recipes including in particular drinks, and in particular vegetable milk analogs. . In addition, as will be exemplified below, the protein composition according to the invention possesses an inhibitory action on DPP-IV which allows it to provide a satietogenic effect during consumption.

[61] More particularly, the invention relates to the application of faba bean isolate in nutritional formulations such as:

- drinks, in particular through mixtures of powders to be reconstituted, in particular for dietetic nutrition (sport, slimming), ready-to-drink drinks for dietetic or clinical nutrition, liquids (drinks or enteral bags) for clinical nutrition vegetable drinks

- yoghurt-type fermented milk (stirred, Greek-style, drinkable, etc.)

- in vegetable creams (such as coffee cream or "coffee whitener"), dessert creams, ice cream desserts or sorbets.

- cookies, muffins, pancakes, nutritional bars (intended for specialized slimming nutrition or for athletes), breads, especially gluten-free breads enriched in proteins, high protein cereals, obtained by extrusion cooking (“crisps” for inclusion, breakfast cereals, “snacks”),

- cheeses,

- meat analogues, fish analogues, sauces, in particular mayonnaise.

[62] The isolate according to the invention is of interest for yoghurts. A yogurt, yogurt or yoghurt is a milk seeded with lactic ferments in order to thicken it and keep it longer. To be called yoghurt, it must contain, and only, two specific ferments, Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus, which give it its specific taste and texture and also provide certain nutritional and health benefits. Other fermented milks (yoghurt-like) have been created in recent years. They may or may not contain these two bacteria, and in addition strains such as Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium bifidum, B. longum, B. infantis and B. brève. Yoghurts are thus an excellent source of probiotics, ie living microorganisms which, when ingested in sufficient quantity, exert positive effects on health, beyond the traditional nutritional effects. Whether firm, stirred or liquid, it retains its name of yogurt, because it is indeed, in addition to the definitions in the Regulations, its manufacture that conditions its final texture. Thus, to obtain a firm yogurt, the milk is directly sown into the pot. While in the case of stirred yoghurt (also known as "Bulgarian"), the milk is inoculated in a tank, then stirred before being poured into its pot. Finally, the liquid yoghurt, also known as drinkable yoghurt, is stirred then beaten until the correct texture is obtained and poured into bottles. But there are also other types of plain yogurts, such as Greek yogurts, with a thicker texture. The percentage of fat can also affect the texture of the yogurt, which can be made from whole, semi-skimmed or skimmed milk (a label containing only the word "yogurt" necessarily means yogurt made with semi-skimmed milk. skimmed). In all cases, its Use By Date (DLC) cannot exceed 30 days and must always be stored in the refrigerator between 0 ° and 6 °.

[63] There are thus three main classes of yoghurt:

- Stirred yoghurt: More liquid, it is often more acidic than plain yogurt.

Only its texture differs. It is also called Bulgarian yogurt - in reference to the supposed origins of yogurt and to Lactobacillus bulgaricus, one of the two ferments at work in the transformation of milk into yoghurt. It is made in vats before being packaged in jars. It is particularly suitable for making drinks, such as lassis, fruit cocktails ...

- Greek yogurt: Particularly thick, it is a very drained natural yogurt (traditional technique) or enriched with cream. Gourmet, very tasty, it is essential for making tsatsiki and for all Eastern European dishes, and quite simply mixed with herbs, it is a delicious aperitif dip. When cold, it can be substituted for thick crème fraîche,

- Drinkable yoghurt: If it exists plain, it is most often sweetened and flavored, and made with beaten stirred yogurt. Conceived in 1974, it allowed teenagers to reconnect with the pleasure of milk, by tasting yogurt without a spoon, directly from the bottle. Recently, it has also been available in “pourable yoghurt”, in a 950 g brick, for those who want to combine cereals and yogurts with breakfast. Low in energy - 52 kcal for a 0% yogurt made from skimmed milk; at 88 kcal for a whole milk yogurt - “plain” yoghurt is naturally not rich in fat and carbohydrates, but contains protein in good quantities. It is also a source of micronutrients (especially calcium and phosphorus) as well as vitamins B2, B5, B12 and A. Made up of 80% water, yoghurt actively participates in the hydration of the body.

[64] Regular consumption of yogurt is thus recognized to improve the digestion and absorption of lactose (opinion of 19 October 2010 from EFSA). Other studies show potential benefits in improving diarrhea in children. children, and the immune system in some people such as the elderly. However, the consumption of cow's milk is increasingly criticized, questioned and we are witnessing a growing number of people who simply decide to eliminate it from their diet, for reasons for example of lactose intolerance, or allergenicity issues. Vegetable milk-based yoghurt solutions have therefore been proposed, because vegetable milks are much more digestible than cow's milk, are rich in vitamins, minerals and unsaturated fatty acids. In the rest of our presentation, for the sake of simplicity, we will continue to use the term "yogurt" even if the origin of the proteins is not dairy (officially, "yogurts" which are made from ingredients other than fermented milk, dairy ingredients, or conventional ferments such as Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus do not have the right to this name). The most common plant source is soy. However, even if soy milk has the greatest richness in calcium and protein, it is also very indigestible; this is why it is not recommended for children. Moreover, it is also not advisable to abuse soy products as their health effects can be counterproductive when consumed in large quantities. Moreover, it is commonly accepted that 70% of world soybean production is GMO.

[65] The isolate according to the invention is also of interest for milk and dairy drinks as well as vegetable drinks. Milk is a food which contains a significant source of protein and of high biological quality. For a long time, animal proteins have been acclaimed for their excellent nutritional qualities because they contain all the essential amino acids in adequate proportions. However, some animal proteins can be allergenic, causing very annoying and even dangerous reactions on a daily basis. One of the most common allergic reactions is an allergy to dairy products. Studies show that 65% of people with food allergies are allergic to milk. The adult form of milk allergy, referred to herein as a “dairy allergy,” is a reaction of the immune system that creates antibodies to fight the unwanted food. This allergy is different from the allergy to cow's milk proteins (proteins cattle), also called APLV, which affects newborns and children. The clinical manifestations of this allergy are mainly gastrointestinal (50 to 80% of cases), also cutaneous (1 0 to 39% of cases) and respiratory (19% of cases). In view of all the disadvantages cited above associated with the consumption of milk proteins, this results in a great interest in the use of replacement proteins, also called alternative proteins, among which are classified plant proteins. Plant milks, obtained from plant ingredients can be an alternative to milks of animal origin. They palliate and avoid the APLV. They are free from casein, lactose, cholesterol, are rich in vitamins and minerals, are also rich in essential fatty acids but low in saturated fatty acids. Some also have interesting fiber levels. Besides the fact that some vegetable milks are poor in calcium, that others because of their botanical rarity are not commercially available, it should also be mentioned that certain vegetable milks are also allergens. This is the case, for example, with vegetable milks prepared from oilseeds, such as, for example, soy milk. In view of all the disadvantages of milk proteins but also of the dangerous allergenic character conferred by certain vegetable proteins, there is a real demand on the part of consumers, unfulfilled to date, for vegetable milks possessing indisputable and recognized harmlessness and being able to therefore be consumed by the whole family. Traditional manufacturers are also starting to look for new sources of protein to enrich their products.

[66] The isolate according to the invention is also of interest for dairy creams for coffee cream, butter, cheese, Chantilly creams, sauces, toppings, cake decoration. Dairy creams are products with more than 30% fat (fat) obtained by concentrating milk, in the form of an emulsion of oil droplets in skimmed milk. They can be used for various applications, either directly as a consumer product (used for example as coffee cream) or as a raw material in industry for the manufacture of other products such as butter, cheese, whipped cream, sauces. , ice creams, or even toppings and cake decorating. There are different varieties of creams: fresh, light, liquid, thick, pasteurized. Creams can be distinguished by their fat content, shelf life and texture. Raw cream is the cream resulting from the separation of milk and cream, directly after skimming and without going through the pasteurization step. It is liquid and contains 30 to 40% fat. Always liquid in texture, pasteurized cream has undergone the pasteurization process. It was therefore heated to 72 ° C. for twenty seconds in order to eliminate microorganisms undesirable for humans. This cream is particularly suitable for expansion. It thus takes on a lighter and more voluminous texture by being beaten to incorporate air bubbles. It is perfect for whipped cream for example. Some liquid creams sold in stores are said to have a long shelf life. They can be stored for several weeks in a cool, dry place. To keep for that long, these creams were either sterilized or heated using the UHT process. For sterilization, the cream is heated for 15 to 20 minutes at 115 ° C. With the UHT (or Ultra High Temperature) process, the cream is heated for 2 seconds at 150 ° C. The cream is then rapidly cooled, which results in better preserving its taste qualities. The cream is naturally liquid, once it is separated from the milk, after skimming. In order for it to take on a thick texture, it goes through the seeding stage. Lactic ferments are thus incorporated which, after maturation, will give the cream this thicker texture and this more acidic and richer taste. Alongside traditional technologies (millennia or centuries old) for obtaining cream from milk, over the last decade, technologies for assembling or reconstituting cream from dairy ingredients have developed. These new technologies for reconstituting dairy creams have obvious advantages in industrial processes, compared to fresh cream: low cost of storage of raw materials, greater flexibility in formulation, independence with regard to the seasonality of the milk. composition of milk. Also, reconstituted dairy creams can benefit from the image of naturalness generally attributed to dairy products, since the regulations require for their manufacture the exclusive use of ingredients. dairy products with or without the addition of drinking water and the same characteristics of the finished product as cream (Codex Alimentarius, 2007). The development of the field of reconstituted dairy creams has opened up new possibilities in the formulation of creams, and more particularly that of the birth of the concept of vegetable creams. Vegetable creams are products similar to dairy creams in which the milk fat is replaced by vegetable fat (Codex Alimentarius, codex Stan 192, 1995). They are formulated from well-defined quantities of water, vegetable fats, milk or vegetable proteins, stabilizers, thickeners and low molecular weight emulsifiers. The physicochemical parameters, such as particle size, rheology, stability and ability to expand are the characteristics which are of primary interest to manufacturers and researchers in the field of the substitution of milk creams by vegetable creams. For example, as in any emulsion, the size of the dispersed droplets (particle size) is a key parameter in the characterization of creams because it has a significant impact, on the one hand, on other physicochemical properties such as rheology and stability, and on the other hand, on sensory properties such as texture and color of creams. The influence of emulsifier type includes both low molecular weight emulsifiers such as mono, diglycerides and phospholipids, as well as high molecular weight ones such as proteins, as well as protein / low molecular weight emulsifier interactions. It is thus known that the concentration of the lipid emulsifier also influences the size of the droplets of creams. In protein stabilized systems, a very high concentration of the lipid emulsifier can lead to a large increase in the average droplet size, due to strong droplet aggregation following desorption of the proteins. The type of proteins used in the formulation can also affect the particle size of the creams. In fact, under the same emulsification conditions, creams based on protein sources rich in caseins, such as skimmed milk powder, generally have average droplet diameters smaller than those based on protein sources rich in protein. whey, such as whey powder. The differences in particle size between creams prepared from the two protein sources (caseins or whey proteins) are linked to the differences in interfacial properties at the oil / water interface, caseins having a greater ability to lower the interfacial tension than whey proteins. Furthermore, the protein concentration in the formulation influences the particle size of the creams. Indeed, it has been shown that at constant mass fraction of oil, the size of the droplets decreases with the protein concentration up to a certain concentration beyond which the size varies very little. The simultaneous presence of the amphiphilic molecules of low molecular weight (surfactant) and high (proteins) in a cream formulation generally results in a decrease in the size of the droplets during emulsification. Furthermore, competitive adsorption at the oil / water interface between surfactants and proteins generally leads, during maturation, to desorption of proteins from the surface of the droplets, which can lead to particle size changes.

Initially, it appears that the emulsification conditions, the choice of ingredients (both protein and lipid) used in the formulation, as well as the temperature, influence the final properties of the creams. It appears that vegetable creams can lead to new technofunctional properties. Thus, the resistance to freezing which can give great stability to ice creams is an example. They can also exhibit stability in hot or cold bonding, which is a considerable advantage, since these creams can be used indifferently in the preparation of hot or cold dishes. If vegetable creams can provide new functionalities and show comparable or even more interesting textural properties than those of dairy creams, the fact remains that they can present sensory defects, in particular with regard to their taste and smell, even sometimes after adding flavorings (case of soy protein, or pea protein).

[67] The isolate according to the invention is also of interest for vegan cheeses. Cheese is normally a food obtained from coagulated milk or dairy cream, followed by draining followed or not by fermentation and possibly refining. The cheese is thus made from cow's milk mainly, but also from sheep, goats, buffalo or other mammals. The milk is acidified, usually using a bacterial culture. An enzyme, rennet, or a substitute such as acetic acid or vinegar, is then added to cause coagulation and form curds and whey. It is known practice to make vegan alternatives to cheese (in particular cheeses of the mozzarella type), by substituting milk caseinates with native and modified starches, more particularly acetate stabilized starches. However, it is still sought to improve the character of “shredding” (Anglo-Saxon term for “shredability”), the melting, the freeze / thaw stability, the flavor (in particular in the United States for pizza preparations). Tests have been carried out by combining oil, modified starches and pea proteins without giving full satisfaction.

[68] The isolate according to the invention is of interest for ice creams. Ice creams conventionally contain animal or vegetable fats, proteins (milk proteins, egg proteins) and / or lactose. Proteins then play the role of texturizer in addition to providing ice cream flavor. They are mainly produced by weighing the ingredients, their pre-mixing, their homogenization, pasteurization, refrigeration at 4 ° C (allowing maturation), then freezing before packaging and storage. Many people, however, suffer from an intolerance to dairy products or other ingredients of animal origin that prevent them from consuming traditional milk or ice cream. For this group of consumers, there is so far no alternative to ice cream containing milk of comparable sensory value. In the heretofore known ice cream preparations having vegetable ingredients, mainly soybean, attempts have been made to replace animal emulsifiers with vegetable proteins. Dried vegetable proteins, obtained in conventional aqueous or hydro-alcoholic extraction processes and after drying in powder form, have often been used. These proteins appear to be heterogeneous mixtures of polypeptides, certain fractions of which have, to varying degrees, particularly good properties as emulsifiers or formation agents. gel, as water binding agents, foam or texture improving agents. Until now, vegetable protein products have been obtained almost exclusively from soybeans, without fractionation according to their specific functional properties. Moreover, the taste of ice creams prepared from said soy proteins is prohibitive.

[69] The isolate according to the invention is of interest for biscuit products, pastry products, bakery products and high-protein cereal products To obtain a “high protein” claim, according to the regulations in force, it is necessary that the protein-related calorie intake is equal to or greater than 20% of the total energy intake of the finished product. This means that, in products containing a substantial fat content such as cookies or cakes (between 10% for the leanest to 25% for the richest with an average of 18% fat), the rate of protein incorporation to achieve the claim is significant and is greater than 20%.

[70] In the field of (total or partial) substitution of milk proteins in food products, it is sought after plant proteins whose functional properties are equivalent, or even improved, compared to milk proteins. The term "functional properties" means in the present application any non-nutritional property which influences the utility of an ingredient in a food product. These various properties contribute to obtaining the desired final characteristics of the dairy product. Some of these functional properties are solubility, viscosity, foaming properties, emulsifying capacities. Proteins also play an important role in the sensory properties of the food matrices in which they are used, and there is a real synergy between the functional properties and the sensory properties. The functional properties of proteins or functionalities are therefore the physical or physico-chemical properties which have an impact on the sensory qualities of food systems generated during technological transformations, preservation or domestic culinary preparations. It can be seen, whatever the origin of the protein, that it affects the color, flavor and / or texture of a product. These organoleptic characteristics play a decisive role in the consumer's choice and in this case they are widely taken into account by manufacturers. The functionality of proteins is the result of their molecular interactions with their environment (other molecules, pH, temperature, etc.). Here, it is a question of the surface properties which group together the properties of interaction of proteins with other polar or non-polar structures in the liquid or gas phase: this covers the emulsifying, foaming properties ...

[71] Within human food applications, the protein composition according to the invention is particularly suitable for dairy applications. More particularly, the invention relates to the application of the feverolle isolate according to the invention for fermented milk of yoghurt type (stirred, Greek style, drinkable) and in dairy or vegetable creams, dessert creams, ice cream desserts or sorbets or cheese.

[72] The nutritional formulations according to the invention can further comprise other ingredients which can modify the chemical, physical, hedonic or processing characteristics of the products or serve as pharmaceutical or complementary nutritional components when they are used for certain target population. . Many of these optional ingredients are known or otherwise suitable for use in other food products and can also be used in nutritional formulations according to the invention, provided that these optional ingredients are safe and effective for oral administration and are suitable for oral administration. compatible with the other essential ingredients of the selected product. Non-limiting examples of such optional ingredients include preservatives, antioxidants, emulsifying agents, buffering agents, pharmaceutical active agents, supplemental nutrients, colors, flavors, thickening agents and stabilizers, etc. Powder or liquid nutritional formulations may further include vitamins or related nutrients, such as vitamin A, vitamin E, vitamin K, thiamine, riboflavin, pyridoxine, vitamin B12, carotendids, niacin , folic acid, pantothenic acid, biotin, vitamin C, choline, inositol, their salts and derivatives, and combinations thereof. Powdered or liquid nutritional formulations may further include minerals, such as phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium, chloride, and combinations thereof. Powdered or liquid nutritional formulations may also include one or more masking agents to reduce, for example, bitter flavors in reconstituted powders. 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. The amount of masking agent in the nutritional powder formulation can vary depending on the particular masking agent selected, the other ingredients of the formulation and other formulation or target product variables.

[73] Said invention will be particularly better understood on reading the following examples.

Examples

[74] Example 1: Comparison of traditional and conventional methods of dehulling external fibers:

[75] The same batch of beans of the Tiffany variety is treated to separate the external fibers and the cotyledons. To do this, two methods are employed.

[76] Method of the prior art: The grains were first treated using a knife mill (SM300, Retsch®) whose speed of rotation was 700 rev / min. The ground material was then treated by turboseparation using a so-called “zig-zag” system (MZM 1 -40, Hosokawa-alpine®). The air speed was 4.0 ms ¹ (23 m ³ .h ¹ ). At the end, a light fraction containing the external fibers and a heavy fraction containing the cotyledons are obtained. The heavy fraction is then ground using a roller mill (MLU 202, Bühler®). Finally, a flour is obtained with a particle size of less than 300 μm (the average size of particles produced using a laser particle size analyzer is 275 μm). The process is shown schematically in FIG. 1.

[77] Improved method according to the invention: The grains were first treated using a stone mill (Alma®). The ground material was then treated by turboseparation using a so-called “zig-zag” system (MZM 1 -40, Hosokawa-alpine®). The air speed was 4.0 ms ¹ (23 m ³ .h ¹ ). At the end, a light fraction containing the external fibers and a heavy fraction containing the cotyledons are obtained. The heavy fraction is then treated using a knife mill (SM300, Retsch®), the rotation of which is 700 rpm and the outlet fitted with a 6 mm grid. The heavy fraction is then ground using a roller mill (MLU 202, Bühler®). Finally, a flour is obtained, the particle size of which is less than 300 μm (the average particle size obtained using a laser granulometer is 285 μm). The process is shown schematically in Figure 2.

[78] Manual separation of the outer fibers (or "hulls") residual in the heavy fraction obtained according to the two methods of the prior art and according to the invention described above is carried out. This consists of taking a 200g sample of the fraction, then manually separating the external fibers still present. These are then weighed (Weight = m). The percentage of residual external fibers is given by the following calculation: (m / 200) * 100

For the method according to the prior art, the percentage is 1.7%. For the process according to the invention, this percentage is reduced to 0.9%.

[79] Example 2a: Production of a protein composition according to the invention

[80] 75 kg of bean flour is prepared with the improved process according to the invention described in paragraph [0077] above. This flour is suspended at 10% by weight of dry matter in drinking water at 20 ° C. The pH is adjusted to 9 by adding potash at 20% by mass (3.4 kg). Homogenization is carried out for 15 minutes, still at 20 ° C. The solution is then sent to a Sedicanter decanter from the Flottweg company (Bowl speed: 60% i.e. 4657 rpm (approximately 3500g), Screw speed at 60% for a Vr = 18.8, Pipette for the supernatant (overflow) at 140 mm, feed at 1 m ³ / h) and the liquid supernatant containing the proteins is recovered.

[81] This supernatant is acidified to pH 4.5 by adding hydrochloric acid at approximately 7% by mass (8.2 kg). It is heated to 60 ° C by injecting steam into a double jacket of the tank, where homogenization is carried out for 15 minutes. The Flottweg Sedicanter is used a second time (Bowl speed at 60%, i.e. 4657 rpm (approximately 3500g) Screw speed at 10% for a Vr = 3.5 up to 40% (Vr = 12.6) Pipette for the overflow at 140 mm initially up to 137 Feed at 700 l / h) but this time to recover the sediment where the coagulated proteins are located.

[82] The sediment is diluted to approximately 15-20% by weight of dry matter and neutralized to pH 7 by adding 20% potash. Heat treatment at 135 ° C is carried out using a nozzle and flash cooling under vacuum at 65 ° C is carried out. The product is finally atomized (inlet temperature of 200 ° C and vapor temperature of 85-90 ° C)

[83] The protein extraction yield from the flour is 86.6%. The protein obtained is called "protein composition according to the invention"

[84] Example 2b: Production of a protein composition according to the invention by wet grinding

[85] The bean seeds were first processed using a stone mill (Alma®). The ground material was then treated by turboseparation using a so-called “zig-zag” system (MZM 1 -40, Hosokawa-alpine®). The air speed was 4.0 ms ¹ (23 m ³ .h ¹ ). At the end, a light fraction containing the external fibers and a heavy fraction containing the cotyledons are obtained. The heavy fraction is then treated using a knife mill (SM300, Retsch®), the rotation of which is 700 rpm and the outlet fitted with a 6 mm grid. The heavy fraction pre-crushed using the knife mill is suspended at 20% by weight of dry matter in drinking water at 20 ° C. The heavy fraction is then crushed using a mill. Hurschel® Comitrol 19300. The pH is adjusted to 9 by adding potash at 20% by mass. Homogenization is carried out for 15 minutes, still at 20 ° C. The solution is then sent to a Sedicanter decanter from the Flottweg company (Bowl speed: 60% i.e. 4657 rpm (approximately 3500g), Screw speed at 60% for a Vr = 18.8, Pipette for the supernatant (overflow) at 140 mm, Feed at 1 m ³ / h) and the liquid supernatant containing the proteins is recovered.

[86] This supernatant is acidified to pH 4.5 by adding hydrochloric acid at approximately 7% by mass. It is heated to 60 ° C by injecting steam into a double jacket of the tank, where homogenization is carried out for 15 minutes. The Flottweg Sedicanter is used a second time (Bowl speed at 60%, i.e. 4657 rpm (approximately 3500g) Screw speed at 10% for a Vr = 3.5 up to 40% (Vr = 12.6) Pipette for the overflow at 140 mm initially up to 137 Feed at 700 l / h) but this time to recover the sediment where the coagulated proteins are located.

[87] The sediment is diluted to approximately 15-20% by weight of dry matter and neutralized to pH 7 by adding 20% potash. Heat treatment at 135 ° C is carried out using a nozzle and flash cooling under vacuum at 65 ° C is carried out. The product is finally atomized (inlet temperature of 200 ° C and vapor temperature of 85-90 ° C)

[88] The protein extraction yield from the flour is 87.8%. The protein obtained is called "protein composition 2b according to the invention"

[89] Example 3: Production of a protein composition according to the prior art

[90] A teaching of Fernandez-Quintela is practiced, (Plant Foods for Human Nutrition, 51, 1997). The beans are first treated with the method of the prior art described in paragraph [0064], then the cotyledons are immersed in water for 10 hours, then dried overnight in an oven at 25 ° C. The cotyledons are then ground into a flour of 300 microns on average. This is suspended in drinking water in a 1/5 water / flour mass ratio and the pH of the solution is rectified to 9.0 with 1N sodium hydroxide. The solution is stirred for 20 min. The insoluble fraction is separated by centrifugation (4000 g / 20 min, 20 ° C) and set aside. The pH of the supernatant is adjusted to pH 4.0 with 1N hydrochloric acid and stirred at 20 ° C for 20 min. The solution is centrifuged (4000 g / 20 min, 20 ° C), and the pellet is lyophilized. We calls this protein composition: "Protein composition according to Example 3 according to the prior art"

[91] Example 4: Comparison of functionalities and analyzes

[92] The different compositions obtained using Examples 2 and 3 are compared from an analytical (dry matter and protein content) and functional (water retention capacity according to test A and staining L) point of view. A commercial protein composition of bean FAVA BEAN PROTEIN ISOLATE 85% is also acquired from the company YANTAI T, FULL BIOTECH CO LTD (lot DFC021606181 / C1377), representative of the bean isolates available on the market. Table 1 below summarizes these analyzes.

[93] [Table 1]

[94] The Table shows the exceptional water retention capacity of the protein composition according to the invention: it is much greater than 3 grams per gram of protein, while the protein compositions according to the invention. the prior art at best barely exceed 2 grams per gram of protein composition.

[95] We can also note an excellent protein richness, greater than 90% for example 2a.

[96] Example 2b is slightly less rich in protein (but remains very rich if compared to pea and soy isolates for example), but its water retention capacity is exceptionally high, 3 times greater than that of the prior art.

[97] Example 4: Nutritional benefit of the protein composition according to the invention:

[98] In this example, it is proposed to present a particular nutritional advantage of the protein composition according to the invention. To do this, as the protein composition of the prior art, use is made, by way of reference, of the commercial pea protein compositions NUTRALYS®, a TUBERMINE® potato protein composition and a PRODIET® milk protein.

[99] We will first of all simulate in vitro gastric and intestinal digestion of said compositions, using the protocol described in “Simulated Gl digestion of dietary protein: Release of new bioactive peptides involved in gut hormone secretion” (Caron & al., in Food Research International, Volume 89, Part 1, 2016, Pages 382-390) The proteins will undergo hydrolysis with pepsin (1/40 weight enzyme / weight protein, pH 3, 2h, 37 ° C) then hydrolysis with pancreatin (1/50 weight enzyme / weight protein, pH 7, 2h, 37 ° C). The inhibitory activity of dipeptidyl peptidase-4 or DPP-IV of the digestates thus obtained is then evaluated. DPP-IV is an enzyme present in cell metabolism, its inhibition leads to a significant increase in the concentration of glucagon-like peptide-1 or GLP-1 (which is an incretin, that is to say an intestinal hormone , secreted by ileum L cells in response to a meal) and glucose-dependent insulinotropic peptide or GIP (which is an enterogastrone secreted by K cells of the duodenum in the postprandial period, potentiating stimulated insulin secretion by glucose in the pancreas). These two hormones cause an increase in secretion insulin and a decrease in the secretion of glucagon, a property that improves the sugar balance in diabetics.

[100] To carry out this evaluation, the following protocol is used which is an adaptation of the protocol described in “Dipeptidyl peptidase-IV inhibitory activity of dairy protein hydrolysates” (Lacroix & Li-Chan, August 2012, International Dairy Journal 25 (2) : 97-102). Briefly, 25 μL of the digestates are placed in test tubes, at concentrations ranging from 1.21 mg.mL ¹ to 13.89 mg.mL ¹ , in order to be pre-incubated with 75 μL of Tris / HCl buffer (100 mM, pH 8.0 ) and 25 µL of DPP-IV (0.018 U.mL ¹ ) at 37 ° C for 5 min in a 96-well microplate. The reaction is initiated by adding 50 μL of Gly-Pro-p-nitroanilide (1 mM). All samples and reagents are diluted in Tris / HCl buffer. The microplate is incubated at 37 ° C. for 1 h, and the absorbance of the released p-nitroanilide is measured at 405 nm every 2 minutes using a microplate reader (ELx808, Biotek, USA). The percentage of DPP-IV inhibition is defined as the percentage of DPP-IV activity, inhibited by a given concentration of a sample (1 mg.mL ¹ ) compared with the response of a control. The percentage inhibition of DPPJV is then plotted against the final concentration of the sample. The IC50 is determined in mg / ml as the final sample concentration causing a 50% inhibition of the activity of DPP-IV, it is expressed in mg / ml. The lower the value of NC50, the better the desired inhibitory activity of the sample.

[101] The results obtained are as follows:

[Table 2]

[102] The inhibitory action of the faba bean protein composition according to the invention is excellent: in fact, its IC50 is halved compared to the proteins of the commercial prior art.

[103] Example 5: So-called “ready-to-drink” or RTD drink with 7% protein:

[104] A so-called "ready-to-drink" or "RTD" drink is produced to compare the bean isolates according to the invention (2a) and a commercial pea isolate NUTRALYS® S85F from the company ROQUETTE.

[105] The receipts are presented in table 3 below:

[106] The process for preparing the drinks is as follows:

- Mixture of different powders

- Heating the water to 50 ° C and introducing the mixture of powders

- Dispersion using a Silverson high shear mixer (30 min, 50 ° C, 3500 rpm)

- Heat the oil to 50 ° C in a separate container, add to the aqueous dispersion and disperse using a Silverson high shear mixer (5 min, 10,000 rpm)

- Heat treatment at 142 ° C for 5 seconds

- High pressure homogenization 200 bars, 2 passes - Cooling to 30 ° C

[107] The different drinks are then compared by analyzing the particle size profile of the emulsion obtained in the drink using a Mastersizer 3000 particle size analyzer (Malvern), measuring the size of the particles by laser diffraction. The sample is measured directly in a liquid way with an optical pattern at 1.50 + 0.01 i. The coefficients well known to those skilled in the art D10, D50, D90 and Dmode are measured to characterize the emulsion of the oil.

[108] By comparing the results, it can be seen that the emulsion obtained with the feverola isolate according to the invention is much smaller, indicating a better emulsion.

[109] Example 6: Plant-based milk or "Milk alternative"

[110] It is proposed here to produce a vegetable milk with the bean isolate 2a according to the invention.

[111] The recipe is as follows:

[112] The preparation protocol is as follows:

- Heat water to 70 ° C and hydrate the protein isolate for 15 min using a Sylverson at 2000 rpm

- Add the other ingredients except the oil and mix for 10 min - Heat the oil to 65 ° C and add with stirring at 6000 rpm

- UHT sterilization 142 ° C 5sec

- Homogenization at 75 ° C 2 stages (270 bars and 30 bars)

- Cooling 4 ° C

[113] This gives a liquid that looks like milk. This vegetable alternative to milk does not undergo any settling during storage.

[114] An analysis of the particle size distribution of the emulsified oil globules was performed using a Mastersizer 3000 particle size analyzer (Malvern). The coefficients describing the particle size distribution are as follows: D10 = 0.19_ microns, D50 = 0.40 microns and D90 = 0.91 microns. These results are excellent and demonstrate an excellent emulsification of lipid globules, just like milk.

[115] Example 7: Classic and light mayonnaise:

[116] We will demonstrate below the excellent results of our isolate 2a according to the invention in the preparation of classic (called "full-fat") and light ("low-fat") mayonnaise.

[117] The ingredients needed to make mayonnaise recipes are as follows:

[118] The isolates to be tested will be Nutralys® F85F from the company ROQUETTE, the bean isolate 2a according to the invention and aquafaba (“Aquafaba Powder” obtained from the company Vôr).

[119] The manufacturing protocol is as follows:

- Mix the ingredients for 1 st phase for 1 min at speed 3 in a HOTMIX Pro Gastro (Manufacturer: MATFER - FLO, Model: 212502).

- Add the ingredients for the 2nd and 3rd phase for 1 min 30 min at speed between 4 and 7 for the Low Fat or add the ingredients for the 2nd phase for 2 min at speed 3 for the Full Fat.

- Add the ingredients for the 3rd phase for 1 min at speed 3 for Full Fat.

- Add the ingredients for the 4th phase for 1 min at speed 3 for Full Fat.

- Finish the emulsion at speed 8 for Low Fat and 3 for Full Fat for

1 min.

[120] We will compare the different mayonnaise obtained using a TA.HDplus texturometer (Company Stable Micro Systems Ltd), allowing us to measure the parameters of firmness, consistency and cohesion. The firmness (g) corresponds to the force to be applied necessary for the geometry (see the “backward ring extrusion” kit described below) to penetrate into the product, the consistency (g. Dry) is a given calculated according to the area under the firmness and cohesion curve (g) corresponds to the force to be applied for the geometry to withdraw from the mayonnaise.

[121] The texturometer is equipped with the “ring backward extrusion” kit which consists of a disc screwed onto the device and 3 plexiglass containers which are filled with mayonnaise. The acquisition is made using the Exponent software with the program designed for the analysis of mayonnaise. The descent of the geometry is carried out at 3mm / s until it reaches the bottom of the container and the rise is carried out at 5mm / s. The software automatically plots a curve as a function of time allowing the parameters to be deduced.

[122] All implementation is clearly explained in the user manual.

[123] The results of “low-fat” mayonnaise are as follows:

[124] The results of 'full-fat' mayonnaise are as follows:

[125] The results obtained show that the mayonnaises obtained with the feverola isolate according to the invention are characterized by excellent texture values, far above the pea isolate or aquafaba.

[126] Example 9: ice cream:

[127] Here, a NUTRALYS® pea protein isolate and the bean isolate according to the invention are compared in an ice cream recipe.

[128] The different ice cream compositions are as follows:

[129] The preparation protocol is as follows:

- Water heating to 60 ° C

- Add and mix for 5 minutes the water and ¾ of the sucrose

- Addition of the stabilizer and the rest of the sucrose, mixing for 5 min

- Addition of the isolate to be tested, mixing for 5 min

- Add coconut oil, mix for 5 min

- Final mix for 20 min at 60 ° C

- High pressure homogenization at 70 ° C 200 bars

- Pasteurization 80 ° C 3 min in a Powerpoint®

- Cooling to 4 ° C

- Maturation overnight in the refrigerator

- Freezing with Tetrapak freezer, targeting 100% overrun

[130] We will compare the efficiency of expansion on the mixture obtained just before pasteurization. The protocol used is as follows:

- Pour 1 liter of the mixture into the bowl of a Kitchenaid®

- Mix at high speed (10) for 6 minutes and pour into a 2 liter test tube

- Immediate measurement of the volume of mixture and foam at T0

- Remeasure after 15 min

[131] The results obtained are as follows:

[132] For the mixture obtained with the isolate according to the invention, the foam is invisible from the start up to 15 min. This is explained by better retention in the mixture containing the isolate according to the invention. This better retained foam will make it possible to obtain an ice cream with a more homogeneous expansion.

[133] Example 10: gelation at acidic pH

[134] Gelation at acidic pH is an important property, especially when making yoghurts or yoghurts, as well as tofu.

[135] A comparative gel strength analysis of pea and faba bean isolates was performed on the TAXT + texturometer after heat treatment and acidification with gluconodeltalactone (GDL).

[136] The powders are hydrated to 15% dry matter in azide water, placed in a water bath at 60 ° C, with stirring for 5 minutes. The solutions are then left under stirring at room temperature overnight. The next day, GDL is added at a rate of 2% w / w. Immediately after addition, each solution is divided into 3 separate jars (in order to triple the gel strength measurements). Acidification is carried out in order to reach a pH of 4.6. The samples are placed in a water bath at 80 ° C. for 2 hours, before being stored overnight in the fridge. The gel strength measurements are then taken the following day.

[137] The characterizations of the gel were carried out at 20 ° C, with a texturometer

TAXT + from Stable Micro Systems Ltd. The parameters are as follows compression mode, geometry: ball punch P0.5S, pre-test speed: 1 mm / sec, test speed: 0.5mm / sec, post-test speed: 10 mm / sec, distance: 15mm, hold time : 60 sec, trigger force: 5g. The force necessary to apply this displacement is recorded and the maximum force required is retained.

[138] The results are as follows:

We can clearly see that with the isolate according to the invention, a gel strength 3 times greater than with the pea protein isolate is obtained. This observation makes it possible to envisage excellent results during the manufacture of fermented products which are alternatives to yoghurts, spoonable or drinkable. For spoonables, the excellent gel strength makes it possible to consider formulations without hydrocolloids such as pectin.

[139] Example 11: Yogurts

[140] We compare a pea protein isolate Nutralys® S85F from the company Roquette and the bean isolate 2a according to the invention.

[141] The formulations of the yogurts are as follows:

[142] The preparation protocol is as follows:

- Hydrate the isolates with water at 55 ° C for 30 min with a Sylverson shaker at 2500 rpm

- Add the other ingredients and mix for 5 min at 6000 rpm

- Two-stage high pressure homogenization (150 bar and 45 bar) at 60 ° C

- Pasteurization 95 ° C 10 min

- Cooling to 42 ° c and addition of YOFLEX® YF-L02DA ferments

- Maintained at 42 ° C for acidification by fermentation until pH 4.6 is obtained

- Homogenization at 4000 rpm with IKA Magic Lab

- Storage 4 days at 4 ° C [143] We compare the firmness of the yogurts obtained using a TAXT + texturometer. The results are as follows:

[144] It can be seen that the bean isolate yogurts are firmer because the force required to perform the analysis is much greater.

Claims

Claims

[Claim 1] Bean protein composition whose color is characterized by an L component greater than 70, preferably greater than 75, even more preferably greater than 80 according to the L * a * b measurement and the water retention according to test A is greater than 3 grams, preferably greater than 3.5 grams of water per gram of isolate.

[Claim 2] Protein composition according to Claim 1, characterized in that its high protein content is greater than 70% by weight expressed as a percentage of proteins on dry matter, preferably greater than 80% by weight, even more preferably greater than 90% by weight. weight.

[Claim 3] Protein composition according to Claims 1 or 2, characterized in that it has a dry matter greater than 80% by weight, preferably greater than 85% by weight, even more preferably greater than 90% by weight.

[Claim 4] A method of producing a protein composition according to one of claims 1 to 3, characterized in that it comprises the following steps:

1) Implementation of bean seeds;

2) Grinding of the beans using a stone mill, followed by separation of the ground material obtained into two so-called light and heavy fractions using an ascending air flow, then a second grinding of the heavy fraction with a knife mill;

3) Final grinding of the heavy fraction using a mill selected from roller mills and knife mills to obtain a flour;

4) Suspending the flour in an aqueous solvent;

5) Removal of the solid fractions from the suspension by centrifugation and obtaining a liquid fraction;

6) Isolation by precipitation by heating at isoelectric pH of the bean proteins contained in the liquid fraction;

7) Dilution of the bean proteins previously obtained to 15-20% by weight of dry matter and neutralization of the pH between 6 and 8, preferably 7, to obtain the protein composition of beans;

8) Drying of the protein composition of faba beans

[Claim 5] A method according to claim 4 characterized in that the average grain size of the flour obtained during step 3 is between 200 and 400 microns, preferably 300 microns.

[Claim 6] A method according to claim 4 to 5 characterized in that the pH of the aqueous solvent during step 4 is adjusted between 8 and 10, preferably 9.

[Claim 7] Process according to one of Claims 4 to 6, characterized in that the temperature of the aqueous solvent in step 4 is adjusted between 2 ° c and 30 ° c, preferably between 10 ° C and 30 ° C, preferably between 15 ° C and 25 ° C, even more preferably at 20 ° C.

[Claim 8] Method according to one of claims 4 to 7, characterized in that the acidification of step 6 is carried out at a pH between 4 and 5, preferably 4.5.

[Claim 9] Method according to one of claims 4 to 8, characterized in that the heating temperature is between 45 ° C and 75 ° C, preferably between 50 ° C and 70 ° C, even more preferably between 55 ° C and 65 ° C, the most preferred being 60 ° C and the heating time is between 5 minutes and 25 minutes, preferably between 10 and 20 minutes, the most preferred being 10 minutes.

[Claim 10] A method according to one of claims 4 to 9 characterized in that step 7 also contains a heat treatment, preferably at a temperature of 135 ° C by direct injection of steam by nozzle and cooling by the flash effect under- vacuum at 65 ° C.

[Claim 11] Method according to one of claims 4 to 10, characterized in that step 8 also contains drying, preferably by multiple effect atomization.

[Claim 12] Method according to one of claims 4 to 11, characterized in that steps 3 and 4 of the method are carried out concomitantly in order to carry out the final grinding of the heavy fraction in the presence of aqueous solvent.

[Claim 13] A method according to claim 12 characterized in that the pH of the aqueous solvent during the final grinding step of the heavy fraction in the presence of aqueous solvent is adjusted between 8 and 10, preferably 9.

[Claim 14] Industrial use, in particular in human or animal food, in cosmetics, in pharmacy, of the faba bean protein composition according to one of claims 1 to 3 or obtained by the process according to one of claims 4 to 13 .

[Claim 15] Use according to claim 14 in:

- drinks, in particular drinks for dietetic or clinical nutrition, drinks or enteral bags, vegetable drinks,

- fermented milk of the yoghurt type,

- vegetable creams,

- dessert creams,

- frozen desserts or sorbets,

- cookies, muffins, pancakes,

- nutritional bars intended for dietetic nutrition

- breads,

- high protein cereals.

- cheeses,.

- meat analogues,

- fish analogues,

- sauces, in particular mayonnaise.