Small broad bean protein composition

Abstract

The present invention relates to the field of legume protein isolates, and particularly to faba bean protein isolates. The present invention also relates to the production method and industrial application of the protein isolates.

Description

Technical Field

[0001] The present invention relates to the field of faba bean protein isolates, and more particularly to faba bean protein isolates from small broad beans. Background Art

[0002] The small broad bean (féverole or féverolle (old spelling)) is an annual plant of the broad bean species (Vicia faba). The small broad bean belongs to the Fabaceae family, Faboideae subfamily, and Fabeae tribe of leguminous plants.

[0003] The small broad bean is of the same species as the broad bean and is a plant that has been consumed by humans since ancient times. Therefore, the term "broad bean" refers to both the seed and the plant.

[0004] In the prior art, various production methods are known for producing protein isolates from small broad bean seeds.

[0005] "Potential of Fava Bean as future protein supply to partially replace meat intake in the human diet" (Multari et al., Comprehensive Reviews in Food Science and Food Safety, Vol. 14, 2015) provides an excellent review of the existing knowledge on this subject.

[0006] The traditional method consists of obtaining a bean flour by grinding small broad beans. It is then diluted in water for alkaline extraction, aiming to dissolve the small broad bean proteins. The solution is then subjected to liquid / solid separation, thereby obtaining on the one hand a crude protein solution and on the other hand a solid fraction rich in starch and fiber. The protein is extracted by isoelectric pH precipitation of the protein, separated from the aqueous solution and dried.

[0007] The protein isolate thus obtained has a protein content of at least 80% (expressed as the ratio of the total nitrogen content multiplied by the factor 6.25 to the total dry matter, calculated as described in the document at the following address: http: / / www.favv-afsca.fgov.be / laboratories / methods / fasfc / _documents / METLFSAL003Proteinebrutev10.pdf). The protein isolate has known long-term industrial benefits, particularly in human and animal food. In fact, its nutritional and functional properties make it suitable for use in many recipes and formulations.

[0008] However, to date, those skilled in the art have had to cope with two major technical problems.

[0009] First, the resulting protein isolate is typically characterized by a dark grey, even black, appearance. This is mainly due to the presence of tannic acid and polyphenols in the external fibers produced by the protein during the manufacturing process of the protein isolate.

[0010] Despite great care, traditional methods of external fiber removal (referred to as "dehulling" in English) do not sufficiently remove tannic acid and polyphenols, and the obvious dark appearance limits the possible range of applications.

[0011] Optimized methods have been developed. For example, the method described in "Technological-scale dehulling process to improve the nutritional value of faba beans" (Meijer et al., Animal Feed Science and Technology, No. 46, 1994) proposes two grindings, two filtrations, and one turbo separation (classifying particles by density using an upward air flow). These technical improvements are complex and expensive.

[0012] Since tannic acid and polyphenols are soluble at alkaline pH values, there is also a strategy of not performing the above-mentioned alkaline extraction. However, if the dissolution of these compounds is limited, thereby limiting the dark appearance, the extraction rate will be severely limited. In fact, since faba bean protein is more soluble at alkaline pH values, extraction at neutral or acidic pH values will limit the extraction rate.

[0013] Secondly, the water retention rate of the faba bean protein isolate obtained according to the prior art is less than 3 grams per gram of protein. The water retention rate is a measure of the amount of water that can be absorbed by the protein isolate after contact with an aqueous solvent under the conditions specified in Test A, as described in detail hereinafter in this specification.

[0014] For example, as described in the article "Nutritional and functional properties of Vicia Fabaprotein isolates related fractions" (Vioque, Food Chemistry, Vol. 132, 2012), the water-holding capacity of the isolate is 2.55 grams of water per gram of protein (see Table 3 of the article). Similarly, as described in "Composition and functional properties of protein isolatesobtained fromcommercial legumes grown in northern Spain" (Fernandez-Quintela, PlantFoods for Human Nutrition, Vol. 51, 1997), the water-holding capacity of the isolate is 1.8 grams of water per gram of protein (see Table 4 of the article).

[0015] These values are applicable to certain industrial applications and may be limited for other applications.

[0016] Therefore, for this technology, it is important to explore a simple and effective method that can make the small broad bean protein as light-colored as possible and have a water retention rate higher than 3 grams of water per gram of isolate.

[0017] The applicant has the privilege of discovering such a method and such isolates. The present invention will be described hereinafter. Summary of the Invention

[0018] According to the present invention, a small broad bean protein composition is provided. The color of the small broad bean protein composition is characterized in that the component L according to the L*a*b measurement method is higher than 70, and the water retention rate is higher than 3 grams of water per gram of isolate.

[0019] According to another aspect, a method for producing a faba bean protein composition according to the present invention is provided, characterized in that the method comprises the following steps: 1) providing faba bean seeds; 2) milling the faba bean seeds using a stone mill, then separating the resulting milled product into two components called light and heavy using an upward air flow, and then subjecting the heavy component to secondary milling using a knife mill; 3) subjecting the heavy component to final milling using a roller mill to obtain a powder; 4) suspending the powder in an aqueous solvent; 5) removing the solid component from the suspension by centrifugation and obtaining a liquid component; 6) heating under isoelectric pH conditions and separating the faba bean protein contained in the liquid component by precipitation; 7) diluting the previously obtained faba bean protein to 15 wt% - 20 wt% of dry matter and neutralizing the pH value to between 6 and 8, preferably 7, to obtain the faba bean protein composition; 8) drying the faba bean protein composition.

[0020] According to the last aspect, an industrial use of a faba bean protein isolate according to the present invention is provided, specifically its use in human or animal food, cosmetics, and pharmaceuticals.

[0021] The present invention and its modifications can generally provide practical and effective solutions to meet the industrial need for obtaining a faba bean protein isolate, the color characteristics of which are such that the component L according to the L*a*b measurement method is higher than 70 and the water retention rate is higher than 3 grams of water per gram of isolate, and also provide a production method and suitable industrial uses thereof.

[0022] The present invention can be better understood through the descriptions in the following sections. Description of the Drawings

[0023] The following will elaborate on other features, details, and advantages of the present invention through detailed descriptions and in conjunction with the following drawings:

[0024] Figure 1

[0025] Figure 1 shows a method for separating the outer fiber and cotyledon of traditional faba bean seeds;

[0026] Figure 2

[0027] Figure 2 shows a method for separating the outer fiber and cotyledon of faba bean seeds according to the present invention. Detailed Description of the Invention

[0028] ​​As described above, according to the present invention, a small broad bean protein composition is first proposed. The characteristics of the color of the small broad bean composition are that the component L according to the L*a*b measurement method is higher than 70, preferably higher than 75, and even more preferably higher than 80, and the water retention rate according to Test A is higher than 3 grams of water per gram of isolate, preferably higher than 3.5 grams of water per gram of isolate.

[0029] "Small broad bean" refers to a leguminous plant belonging to the legume family, Papilionoideae subfamily, and Vicia tribe, and is an annual plant of the broad bean species. It is divided into "minor" and "major" varieties. In the present invention, wild varieties and varieties obtained by genetic engineering or variety selection are excellent sources of varieties.

[0030] "Protein composition" refers to any protein-rich composition obtained by plant extraction and, if necessary, by purification. A concentrate with a protein content higher than 50% of the dry matter is distinguished from an isolate with a protein content higher than 80% of the dry matter.

[0031] "L*a*b measurement" refers to the color evaluation carried out using an appropriate spectrophotometer according to the color space method proposed by the CIE (International Commission on Illumination) in the publication "Colorimetry" (2nd edition, 1986, page 36, article 15). This method converts the color evaluation into three parameters: the value of the brightness L ranges between 0 (black) and 100 (reference white); the parameter a represents the value on the green → red axis, and the parameter b represents the value on the blue → yellow axis. This color measurement is preferably carried out using a DATA COLOR–DATA FLASH 100 or KONIKA MINOLTA CM5 spectrophotometer and implemented with the help of its user manual.

[0032] "Water retention rate" refers to the number of grams of water that one gram of protein can absorb.

[0033] To measure this water retention capacity, Test A with the following protocol is adopted.

[0034] Weigh 20 g of the sample to be analyzed with a beaker, add drinking water at room temperature (20°C ± 1°C) until the sample is completely immersed, let it stand in contact for 30 minutes, separate the residual water and the sample with a sieve, and weigh the final weight of the rehydrated sample (unit: gram).

[0035] Then calculate to obtain the water retention capacity (unit: g) = (P - 20) / 20

[0036] Preferably, the isolate according to the present invention is characterized in that the protein content expressed as a weight percentage of protein in the dry matter is higher than 70%, preferably higher than 80%, and even more preferably higher than 90%.

[0037] Preferably, the dry matter of the protein composition according to the present invention is higher than 80% by weight, preferably higher than 85% by weight, and even more preferably higher than 90% by weight. Any method for measuring water content can be used to quantify such dry matter, preferably gravimetric techniques for assessing water loss by drying.

[0038] It consists of determining the amount of water evaporated by heating a known quantity of a sample of known mass.

[0039] - First, weigh the sample and measure the mass m1 (unit: g).

[0040] - Place the sample in a heating chamber to evaporate the water until the sample mass stabilizes, where the water is completely evaporated. Preferably, the temperature at atmospheric pressure is 105 °C.

[0041] - Weigh the final sample and measure the mass m2 (unit: g).

[0042] - Dry matter = (m2 / m1) * 100.

[0043] A second aspect of the present invention lies in a method for producing a faba bean protein composition according to the present invention, characterized in that the method comprises the following steps: 1) providing faba bean seeds; 2) grinding the faba bean seeds using a stone mill, and then separating the resulting ground material into two components called light and heavy using an upward air flow, and then subjecting the heavy component to secondary grinding using a knife mill; 3) subjecting the heavy component to final grinding using a mill selected from a roller mill and a knife mill to obtain a powder; 4) suspending the powder in an aqueous solvent; 5) removing the solid components from the suspension by centrifugation and obtaining a liquid component; 6) heating under isoelectric pH conditions and separating the faba bean protein contained in the liquid component by precipitation; 7) diluting the previously obtained faba bean protein to 15% - 20% by weight of dry matter and neutralizing the pH value to between 6 and 8, preferably 7, to obtain the faba bean protein composition; 8) drying the faba bean protein composition.

[0044] "Stone mill" refers to a system consisting of two superimposed stone columns, where the space left between the stone columns is approximately equal to the seed particle size. One of the cylinders is stationary, while the other cylinder is rotating. Seeds are added between these two cylinders, and their relative movement will exert physical stress on these seeds.

[0045] "Knife mill" refers to a system consisting of a chamber equipped with an upper inlet for adding seeds, a plurality of knives provided on a rotating shaft disposed in the chamber, and a lower outlet that allows only seeds of the desired particle size to flow out.

[0046] The first step consists in providing small broad bean seeds. These small broad bean seeds also include their protective outer fibers, also known as "hulls" in English. The seeds can be subjected to the necessary pre-treatments, including steps such as cleaning, sieving (e.g., separating the seeds from pebbles), soaking, bleaching or baking. Preferably, if bleaching is carried out, the heat treatment conditions are 3 minutes at 80 °C. Non-limiting examples of varieties include, for example, the Tiffany, FFS or YYY varieties. Preferably, small broad bean varieties with a naturally low tannin and / or polyphenol content will be used, such as the Organdi variety. Such varieties are known and can be obtained by varietal crossing and / or genetic modification.

[0047] The purpose of the second step is to separate the outer fibers and the cotyledons as effectively as possible. First, the small broad bean seeds are milled once using a stone mill. A particularly suitable specific example of such a stone mill is the stone mill sold by the company. As mentioned above, the seeds will be introduced into the space formed by two stone disks, one of which is rotating. The applicant has noticed that this technique is particularly useful as it very effectively separates the outer fibers and the cotyledons of the seeds. Preferably, the space between the stone disks is adjusted to be between 0.4 mm and 0.6 mm.

[0048] Then the obtained milled product is subjected to the action of a countercurrent upward air flow. Different solid particles will be classified according to their density. Generally, two components are obtained after stabilization: a light component mainly containing the outer fibers or "hulls" and a "heavy" component mainly containing the cotyledons. A particularly suitable specific example of the appropriate equipment is the MZMZ1-40 sold by the company.

[0049] Then the heavy component rich in cotyledons will be milled using a knife mill. A particularly suitable specific example of such a stone mill is the SM300 sold by the company.

[0050] The purpose of continuously performing the above three operations in the second step is to very finely separate the outer fibers and the cotyledons while avoiding damage and mixing of these two parts. The methods of the prior art are either too simple to effectively separate the outer layer of fibers or very complex and thus difficult to operate from an industrial perspective. For example, the method described in "Technological-scale dehulling process to improve the nutritional value of faba beans" (Meijer et al., Animal Feed Science and Technology, 46, 1994) proposes two grindings, two filtrations, and one turbo separation (using an upward air flow). This method can obtain a cotyledon component that still contains 1.2% of the outer fibers. Our invention simplifies this method (using different types of mills with different technologies for the two grindings and performing a turbo separation between the two grindings) and can reduce the content of the outer fibers to a value of 1% or lower.

[0051] The purpose of the third step is to grind using a mill selected from a roller mill and a knife mill, especially using a roller mill, to reduce the particle size of the heavy part rich in cotyledons by grinding. Such a roller mill, called "dry" grinding, i.e., solvent-free grinding, a particularly suitable specific example of which is the MLU 202 sold by the company. It is used here to reduce the overall particle size of the powder, thereby obtaining a uniform and sufficiently fine powder to facilitate the subsequent step 4. The preferred particle size is between 200 microns and 400 microns, more preferably 300 microns. To measure this particle size, it is preferably measured using a laser particle size analyzer, but any method is feasible, such as sieving.

[0052] Optionally, the step of reducing the particle size of the heavy component rich in cotyledons, also called the final grinding of the heavy component, can be carried out in the presence of an aqueous solvent, preferably water. In this case, the following fourth step is combined with the third step and then implemented simultaneously. In this case, a suitable mill is, for example, the Comitrol 3000 knife mill.

[0053] The purpose of the fourth step is to suspend the powder obtained in the previous third step in an aqueous solvent, preferably water. Here, the purpose is to selectively extract them by dissolving certain compounds, mainly proteins as well as salts and sugars. The pH value of the solution is advantageously adjusted to an alkaline pH value so as to maximize the dissolution of proteins. This pH value adjustment can be carried out before and / or after the powder is suspended in the aqueous solvent.

[0054] The aqueous solvent is preferably water. However, compounds that can promote dissolution, for example, can be added to the aqueous solvent. The pH value of the aqueous solvent is adjusted to be between 8 and 10, preferably 9. Any alkaline reagent such as soda or lime can be considered, but potassium hydroxide is preferred. The temperature is adjusted to be between 2°C and 30°C, preferably between 10°C and 30°C, preferably between 15°C and 25°C, and even more preferably 20°C. This temperature needs to be adjusted throughout the extraction reaction process.

[0055] The alkaline pH value can effectively maximize the dissolution of proteins. Unfortunately, tannic acid and / or polyphenols also dissolve at alkaline pH values. Some broad bean extraction methods avoid this alkaline pH adjustment, which is beneficial for limiting the contamination effect caused by polyphenols. The specific method we implement in the second step can achieve this alkaline extraction without over-dissolving polyphenols.

[0056] The obtained powder is diluted to obtain a suspension with the powder weight percentage in the total weight of the water / powder suspension being between 5% and 25%, preferably between 5% and 15%, preferably between 7% and 13%, and even more preferably between 9% and 11%, and most preferably 10%. The suspension is stirred using any equipment well-known to those skilled in the art, such as a container equipped with a stirrer, equipped with blades, marine paddles, or any effective fermentation equipment. The extraction time, preferably the extraction time with stirring simultaneously, is between 5 minutes and 25 minutes, preferably between 10 minutes and 20 minutes, and even more preferably 15 minutes.

[0057] The purpose of the fifth step is to centrifuge the soluble components or solid components obtained in the fourth step. The preferred industrial principle can be found in European Patent Application EP1400537, which is hereby incorporated by reference. The principle of this method is to first use a hydrocyclone to extract the starch-rich components, and then use a horizontal decanter to extract the inner fiber-rich components. However, an industrial centrifuge can also be used to extract the starch-rich and inner fiber-rich components. In any case, solid components and a liquid component concentrated with most of the proteins are obtained.

[0058] The purpose of the sixth step is to acidify the broad bean protein to an isoelectric pH value of about 4.5, and then heat the solution to coagulate the protein called globulin, which is separated by centrifugation.

[0059] Acidify to a pH value between 4 and 5, preferably 4.5. Preferably, this is carried out using hydrochloric acid at a mass percentage of about 7%, but any type of acid, inorganic or organic acid, such as citric acid, can also be used. Even more preferably, it is also feasible to use pure ascorbic acid or in combination with other inorganic or organic acids. Acidification with ascorbic acid can improve the final color development. Any subsequent heating method is feasible, such as through a stirred tank equipped with a jacket and / or coil or an in-line steam jet cooker (English "jet cooker"). The heating temperature is advantageously between 45 °C and 75 °C, preferably between 50 °C and 70 °C, even more preferably between 55 °C and 65 °C, and most preferably 60 °C. The heating time is advantageously between 5 minutes and 25 minutes, preferably between 10 and 20 minutes, and most preferably 10 minutes.

[0060] The protein composition mainly composed of globulin will coagulate and precipitate in the solution. It can be separated by any centrifugation technique, such as Sedicanteur. The resulting residual solution is concentrated with sugar, salt, and albumin and is called broad bean solubles. It is treated separately, preferably by evaporation and / or drying.

[0061] It should be noted that the prior art for extracting proteins from broad beans only uses isoelectric precipitation without heating. By combining the two steps according to the present invention, the isolate according to the present invention can be obtained, and broad bean solubles (the name of the supernatant obtained after precipitation and centrifugation) that are stable to temperature can also be obtained. In fact, when the broad bean solubles obtained by isoelectric precipitation are exposed to high temperatures, such as in an evaporator, precipitation will occur. This precipitate becomes a major drawback because it causes blockage of industrial facilities.

[0062] On the other hand, the combination of isoelectric precipitation and controlled heating proposed by the present invention can obtain:

[0063] - Coagulated protein flocs, which after the required treatment result in the product claimed in this application, and

[0064] - Residual solubles containing other soluble proteins (albumin), salts, sugars, etc.

[0065] The second component can generally be utilized in the fermentation and / or animal nutrition industries. For this purpose, it needs to be concentrated to be stable from a bacteriological perspective. For this purpose, the traditional operation is to concentrate by vacuum evaporation, which is completed using a secondary heating different from that which causes the flocs to coagulate. In this operation, as well as in the case of simple isoelectric precipitation during the floc / soluble separation process, the coagulated protein deposits will accumulate in the evaporator.

[0066] In the seventh step, the protein composition is subsequently diluted to about 15 wt% - 20 wt% dry matter and neutralized to a pH value between 6 and 8, preferably 7, using any type of alkali agent, preferably 20 wt% potassium hydroxide.

[0067] Subsequently, the protein composition can be heat-treated, preferably by direct injection of steam through a nozzle at a temperature of 135 °C and cooled by flash evaporation under vacuum at 65 °C.

[0068] The resulting protein composition can be used directly, for example, by hydrolysis with proteases or by texturization through an extruder.

[0069] In the eighth step, the protein composition according to the invention is dried. The preferred drying method is atomization, specifically using a multi-effect atomizer. Typical parameters are an inlet temperature of 200 °C and a vapor temperature of 85 - 90 °C.

[0070] According to a last aspect, an industrial use of the faba bean protein isolate according to the invention is proposed, specifically in human or animal food, cosmetics, and pharmaceuticals. The faba bean composition obtained according to the invention has a very high protein content and a very white color, and thus can be added to many recipes, including specifically beverages, including specifically plant milk analogues. Additionally, as will be illustrated below, the protein composition according to the invention has DPP-IV inhibitory activity, which makes it possible to produce a feeling of satiety after consumption.

[0071] More specifically, the present invention relates to the use of the faba bean isolate in nutritional formulations, such as:

[0072] - Beverages, especially those obtained by mixing reconstitutable powders, mainly for dietary nutrition (sports, weight loss), ready-to-drink beverages for dietary or clinical nutrition, clinical nutrient solutions (beverages or enteral bags), plant-based beverages,

[0073] - Yogurt-type fermented milks (stirred, Greek yogurt, drinking-type, etc.),

[0074] - Plant-based creams (such as coffee creamers or "coffee whiteners"), dessert creams, ice cream desserts or sorbets,

[0075] - Biscuits, muffins, pancakes, nutritional bars (specifically for weight loss or nutrition for athletes), bread, especially gluten-free bread rich in protein, high-protein cereals obtained by extrusion cooking (including "chips", breakfast cereals, "snacks"),

[0076] - Cheeses,

[0077] - Meat analogues, fish analogues, sauces, specifically mayonnaise.

[0078] The isolates according to the present invention can be used in yogurt. Yogurt (yogourt or yoghourt) is a dairy product that is inoculated with a lactic acid starter culture, which thickens it and allows it to be preserved for a longer time. So-called yogurt must contain and only contain two specific starter cultures, namely Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus, which give yogurt its special taste, texture, and also provide certain nutritional and health benefits. In recent years, other fermented milks (yogurt-like textures) have been introduced. They may or may not contain these two bacteria, and also contain strains such as Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium bifidum, B. longum, B. infantis, and B. breve. Therefore, yogurt is an excellent probiotic, i.e., a source of live microorganisms that, when ingested in sufficient amounts, has a positive effect on health, superior to its traditional nutritional role. Whether it is fermented, stirred, or liquid yogurt, it retains the name yogurt. In addition to the definition in the regulations, this is actually because of the manufacturing process that determines its final texture. Thus, to obtain fermented yogurt, milk is directly inoculated in a tank. For stirred yogurt (also known as "Bulgarian yogurt"), the milk is inoculated in a container and then stirred before being canned. Finally, liquid yogurt, also known as drinking yogurt, is stirred and then broken up until the appropriate texture is obtained and poured into bottles. However, there are also other types of plain yogurt, such as Greek yogurt, which has a thicker texture. The fat percentage also affects the texture of yogurt, and the yogurt can be made from whole, semi-skimmed, or skimmed dairy products (labels that only contain the word "yogurt" must mean that the yogurt is made from semi-skimmed milk). In any case, its shelf life (D.L.C) must not exceed 30 days, and it must always be stored in a refrigerator between 0°C and 6°C.

[0079] Therefore, yogurt is divided into three main categories:

[0080] - Stirred yogurt: Thinner, it is usually more acidic than plain yogurt. The difference lies only in its texture. Also known as Bulgarian yogurt - referring to the origin of yogurt and the presumption of Lactobacillus bulgaricus, Lactobacillus bulgaricus is one of the two starter cultures that convert milk into yogurt. It is made in a container and then canned. It is particularly suitable for making beverages, such as lassi, fruit cocktails, etc.

[0081] - Greek yogurt: Extra thick, this is an extremely thick (traditional technology) or cream-rich plain yogurt. A delicacy, very creamy, it is essential for making Greek yogurt with cucumber and all Eastern European dishes, and can be simply mixed with herbs to make a delicious aperitif. When cooled, it can replace thick whipped cream.

[0082] - Drinkable yogurt: If plain, it is usually sweetened and flavored, and is made from whipped stirred yogurt. It was once envisioned in 1974 that it could allow teenagers to rediscover the pleasure of drinking milk, and drink yogurt directly from a bottle without a spoon. Recently, "pourable yogurt" has also been launched, in 950g square boxes, suitable for people who like to pair yogurt with cereals for breakfast. Yogurt made from low-calorie - skim milk is 52kcal; yogurt made from whole milk is 88kcal - the fat and carbohydrate content in "plain" yogurt is naturally not high, but it contains a large amount of protein. It is also a source of micronutrients (especially calcium and phosphorus) and vitamins B2, B5, B12, A.

[0083] Yogurt composed of 80% water plays an active role in human hydration.

[0084] Therefore, regular consumption of yogurt is considered to improve the digestion and absorption of lactose (EFSA opinion on October 19, 2010). Other studies have shown potential benefits for improving childhood diarrhea and the immune system in some populations (such as the elderly). However, the consumption of milk is increasingly being criticized and questioned, and we are seeing more and more people simply decide to remove milk from their diet due to reasons such as lactose intolerance or allergy problems. Therefore, yogurt solutions based on plant milk have been proposed because plant milk is more digestible than milk and is rich in vitamins, minerals, and unsaturated fatty acids. In the following elaboration, for simplicity, even if the source of protein is not milk, we will continue to use the term "yogurt" (formally, "yaourts" refers to those made using ingredients other than fermented milk, milk components, or traditional starters such as Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus, and this name cannot be used). The most commonly used vegetable source is soybeans. However, even though soy milk has the highest calcium and protein content, it is extremely difficult to digest; this is why it is not recommended for children. Additionally, it is not recommended to consume too many soy products because excessive consumption can be counterproductive to health. Moreover, it is generally believed that 70% of the global soybean production is genetically modified crops.

[0085] The isolates according to the invention can also be used in dairy products, milk beverages and vegetable beverages. Milk is a food containing an important source of high-quality biological protein. For a long time, animal proteins have been highly regarded for their excellent nutritional quality because they contain an appropriate proportion of all essential amino acids. However, some animal proteins may cause allergies and can cause extremely uncomfortable reactions, even be dangerous, when consumed daily. Dairy allergy is one of the most common allergic reactions. Studies have shown that 65% of food allergy sufferers are allergic to milk. Adult-onset milk allergy, referred to herein as "dairy allergy", is a reaction of the immune system to produce antibodies against unwanted foods. This allergy is different from milk protein allergy (cow protein), also known as APLV, which affects newborns and children. The clinical manifestations of this allergy are mainly gastrointestinal (50% to 80% of cases), skin (10% to 39% of cases) and respiratory (19% of cases). Given all the above disadvantages associated with the consumption of milk proteins, the use of protein substitutes, also known as alternative proteins, which include vegetable proteins, is of great significance. Plant milks obtained from plant ingredients can replace animal milk. They alleviate and avoid APLV. They do not contain casein, lactose, cholesterol, are rich in vitamins and minerals, and are also rich in essential fatty acids, but have a low saturated fatty acid content. Some also have a high fiber level. Apart from the fact that some plant milks have a low calcium content and others are not available on the market due to the rarity of the plants, it should also be noted that some plant milks can also cause allergies. For example, this is the case for plant milks prepared from oil crops, such as soy milk. Given all the disadvantages of milk proteins and the dangerous allergenic characteristics of some vegetable proteins, the actual demand of consumers for a plant milk with indisputable and recognized harmlessness and suitable for the whole family has not been met. Traditional manufacturers have also started to seek new protein sources to enrich their products.

[0086] The isolate according to the invention is also used for animal cream, used in coffee creamer, butter, cheese, Chantilly cream, sauces, toppings, cake decoration. Animal cream is a product with a fat (MG) content of more than 30%, obtained by concentrating milk, in the form of an emulsion in skimmed milk in the form of oil droplets. They can be used in various applications, either directly as a consumer product (for example as coffee creamer) or as an industrial raw material to make other products, such as butter, cheese, Chantilly cream, sauces, ice cream or even toppings and cake decorations. There are various types of cream: fresh cream, light cream, liquid cream, thick cream, pasteurized cream. Cream can be distinguished by its fat content, storage and texture. Raw cream is cream that is skimmed directly after separating milk and cream without going through a pasteurization step. It is liquid and contains 30% to 40% fat. Pasteurized cream always has a liquid texture and has undergone a pasteurization process. Therefore, it is heated at 72°C for 20 seconds to kill microorganisms that are harmful to humans. This cream is particularly suitable for whipping. The addition of air bubbles gives it a lighter, richer texture. It is ideal for making Chantilly cream, for example. Some liquid creams sold in stores claim to be long-lasting. They can be stored for weeks in a cool, dry place. To be able to keep this long, these creams are either sterilized or heated using the UHT process. In the case of sterilization, the cream is heated at 115°C for 15 to 20 minutes. With the UHT (ultra-high temperature) process, the cream is heated at 150°C for 2 seconds. The cream is then quickly cooled, which better preserves its taste qualities. Cream is naturally liquid after it has been separated from the milk after skimming. To give it a thick texture, it undergoes an inoculation step. Lactic acid starters are then added, which, when matured, give the cream a thicker texture and a more sour, richer taste. In addition to the traditional technology of obtaining cream from milk, which has been around for thousands of years, the technology of combining or reconstructing cream with milk-containing ingredients has also developed in the past decade. Compared to fresh cream, these new animal cream reconstitution technologies offer clear advantages in industrial processes: low raw material storage costs, greater formulation flexibility, and no dependence on the seasonality of milk ingredients. Moreover, reconstituted animal cream can benefit from the natural image usually associated with dairy products, since regulations require that only milk ingredients be used in its manufacture, with or without the addition of drinking water and that the finished product has the same characteristics as cream (Food Law 2007 edition). Developments in the field of reconstituted animal cream have opened up new possibilities for cream formulations, especially the birth of the concept of vegetable cream. Vegetable cream is a product similar to animal cream, in which milk fat is replaced by vegetable fat (Food Law, Standardization Law, 192, 1995 edition). They are formulated with defined amounts of water, vegetable fat, milk protein or vegetable protein, stabilizers, thickeners and low molecular weight emulsifiers. Physicochemical parameters, such as particle size, rheology, stability and swelling amplitude, are the properties that are of greatest interest to manufacturers and researchers in the field of replacing animal cream with vegetable cream.For example, as with any emulsion, the size (particle size) of the dispersed droplets is a key parameter in the characterization of cream, as on the one hand it has a significant impact on other physicochemical properties such as rheology and stability, and on the other hand, it also affects sensory properties such as the texture and color of the cream. The effects of emulsifier type include small molecular weight emulsifiers such as monoglycerides, diglycerides, and phospholipids, as well as high molecular weight emulsifiers such as proteins, and the interaction between proteins / small molecular weight emulsifiers. It is thus known that the concentration of lipid emulsifiers also affects the droplet size of the cream. In a system stabilized by proteins, an extremely high concentration of lipid emulsifiers can cause a significant increase in the average droplet size due to the high aggregation of droplets after protein desorption. The type of protein used in the formulation also affects the particle size of the cream. In fact, under the same emulsification conditions, the average diameter of the droplets in cream based on a protein source rich in casein, such as skim milk powder, is generally smaller than that in cream based on a protein source rich in protein, such as whey powder. The difference in particle size between creams prepared from two protein sources (casein or whey protein) is related to the difference in interfacial properties at the oil / water interface, where the ability of casein to reduce the interfacial tension is higher than that of whey protein. In addition, the protein concentration in the formulation affects the particle size of the cream. In fact, it has been shown that at a constant mass fraction of oil, the droplet size decreases with increasing protein concentration and becomes less variable after reaching a certain concentration. The simultaneous presence of small molecular weight (surfactants) and large molecular weight (proteins) amphiphiles in the cream formulation usually results in smaller droplets during the emulsification process. In addition, competitive adsorption at the oil / water interface between the surfactant and the protein usually leads to protein desorption from the droplet surface during the ripening process, which causes a change in particle size.

[0087] Initially, the emulsification conditions, the choice of ingredients (proteins and lipids) used in the formulation, and the temperature seem to affect the final properties of the cream. It seems that vegetable creams can bring new technological functional properties. Thus, the ability to confer high stability anti-freezing properties to ice cream is an example. They can also have cold or heat adhesion stability, which is a considerable advantage as these creams can be used indifferently for the preparation of hot or cold dishes. If vegetable creams can bring new functions and exhibit texture properties close to or even superior to those of animal creams, they may still have sensory defects, especially in terms of taste and odor, even sometimes after the addition of flavorings (such as soy protein or pea protein).

[0088] The isolates according to the invention can also be used in vegetarian cheese. Cheese, a food product, is typically obtained from coagulated milk or cream, drained, and then fermented or not, and refined if necessary. Thus, cheese is mainly made from cow's milk, but can also be made from sheep's milk, goat's milk, buffalo milk, or other mammalian milk. Cow's milk is acidified using a bacterial culture. Then, an enzyme, rennet, or an alternative such as acetic acid or vinegar is added to cause coagulation and form curds and whey. It is known to produce vegetarian alternatives to cheese (especially of the mozzarella type) by replacing milk caseinate with natural and modified starches, more particularly acetate-stabilized starches. However, there is still a search for improving characteristics such as "shredability", melting, freeze / thaw stability, flavor (especially for pizza preparation in the United States), etc. Experiments have been carried out with oils, modified starches, and pea protein, but they are not entirely satisfactory.

[0089] The isolates according to the invention can be used in ice cream. Ice cream typically contains animal or vegetable fats, proteins (milk proteins, egg proteins), and / or lactose. In addition to flavoring the ice cream, the proteins also act as stabilizers. Their production mainly includes ingredient weighing, premixing, homogenization, pasteurization, refrigeration at 4 °C (to mature it), and then freezing, repackaging, and storage. However, many people are intolerant to dairy products or other animal-derived ingredients, which prevents them from consuming traditional dairy products or ice cream. For such consumers, so far, there has been no alternative except lactose-free ice cream with similar sensory properties. In the preparation of ice cream containing plant ingredients (mainly based on soybeans) known so far, attempts have been made to replace animal emulsifiers with plant proteins. Plant proteins obtained by conventional water or hydroalcoholic extraction methods and dried to a powder form after drying are often used. These proteins belong to a heterogeneous mixture of polypeptides, and some of their components, as emulsifiers or gel-forming agents, as water-binding agents, foaming agents, or texture improvers, have particularly good properties to varying degrees. So far, almost all plant protein products have been obtained from soybeans and have not been fractionated according to their specific functional properties. In addition, the taste of ice cream prepared using the said soy protein is unacceptable.

[0090] The isolates according to the invention are used in biscuit products, pastry products, baked products, and high-protein cereal products. To meet the "protein-rich" standard, according to current regulations, the calories provided by protein must be equal to or higher than 20% of the total calorie value of the finished product. This means that in products with a high fat content, such as biscuits or cakes (with a minimum calorie of 10% to a maximum calorie of 25%, and an average fat content of 18%), to meet this standard, a higher amount of protein is added and is higher than 20%.

[0091] In the field of food (total or partial) milk protein replacement, plant proteins with functional properties equivalent or even improved compared to milk proteins are being sought. The term "functional properties" in this application refers to any non-nutritional property that affects the role of a component in food. These different properties contribute to obtaining the final characteristics required for dairy products. Some of these functional properties are solubility, viscosity, foaming properties, emulsifying capacity. Proteins also play an important role in the sensory properties of the food matrix in which they are used, and there is a real synergy between functional properties and sensory properties. Thus, the functional properties or functions of proteins are physical or physico-chemical properties that affect the sensory quality of food systems generated during technological transformation, storage or home cooking preparation. It can be seen that regardless of the source of the protein, it affects the color, taste and / or texture of the product. These sensory characteristics play a decisive role in the consumer's choice, and in this case, they are generally within the manufacturer's considerations. The function of a protein is the result of the interaction of its molecules with its environment (other molecules, pH, temperature, etc.). Here are surface properties, which include the properties of the protein interacting with other polar or non-polar structures in the liquid or gas phase: including emulsification, foaming properties, etc.

[0092] In human food applications, the protein compositions according to the invention are particularly suitable for dairy applications. More specifically, the present invention relates to the use of the faba bean isolate according to the invention in fermented milks of the yogurt type (stirred, Greek yogurt, drinking type) and animal or vegetable creams, dessert creams, ice cream desserts or sorbets or cheeses.

[0093] The nutritional formulation according to the present invention may also contain other ingredients which can modify the chemical, physical, sensory or processing characteristics of the product, or act as nutraceutical or supplementary ingredients for certain target populations. Many of these optional ingredients are known or otherwise used in other foods and can also be used in the nutritional formulation according to the present invention, provided that these optional ingredients are safe and effective for oral administration and are compatible with other essential ingredients or the selected product. Non-limiting examples of such optional ingredients include preservatives, antioxidants, emulsifiers, buffers, pharmaceutically active agents, supplementary nutrients, pigments, flavors, thickeners and stabilizers, etc. The nutritional formulation in powder or liquid form may also contain 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, their salts and derivatives, and combinations thereof. The nutritional formulation in powder or liquid form may also contain minerals such as phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium, chlorides and combinations thereof. The nutritional formulation in powder or liquid form may also contain one or more flavoring agents to reduce bitterness, for example, in reconstituted powders. Suitable flavoring agents include natural and artificial sweeteners, sodium sources such as sodium chloride, hydrocolloids such as guar gum, xanthan gum, carrageenan and combinations thereof. The amount of flavoring agent in the powdered nutritional formulation may vary depending on the specific flavoring agent selected, other ingredients in the formulation, and other variables of the formulation or target product.

[0094] The present invention will be better understood by the following examples.

[0095] Example

[0096] Example 1: Control of Traditional Conventional External Fiber Removal Method :

[0097] Seeds of the same batch of small broad bean variety Tiffany were processed to separate the outer fibers and cotyledons. For this purpose, two methods were employed.

[0098] Method in the prior art: The seeds were first processed with a knife mill (SM300, ) rotating at 700 rpm. Subsequently, the milled material was processed by turbo separation using a system called "serrated" (MZM 1-40, ). The air flow rate was 4.0 m.s -1 (23 m 3 .h -1 ). Finally, a light component containing outer fibers and a heavy component containing cotyledons were obtained. Subsequently, the heavy component was ground with a roller mill (MLU 202, ). Finally, a powder with a particle size less than 300 μm was obtained (the average particle size obtained using a laser particle size analyzer was 275 μm). This method is asFigure 1 as shown

[0099] According to the improved method of the present invention: The seeds are first processed with a stone mill and then the milled product is processed by turbo separation using a system called "sawtooth" (MZM 1-40, ). The air flow rate is 4.0 m.s -1 (23 m 3 .h -1 ). Finally, a light component containing external fibers and a heavy component containing cotyledons are obtained. Subsequently, the heavy component is processed with a knife mill (SM300, ) with a rotational speed of 700 rpm and an outlet equipped with a 6 mm mesh sieve. Subsequently, the heavy component is ground with a roller mill (MLU 202, ). Finally, a powder with a particle size less than 300 μm is obtained (the average particle size obtained by laser particle size analyzer is 285 μm). This method is as Figure 2 shown

[0100] For two methods according to the prior art and the heavy components obtained according to the present invention above, the residual external fibers (or "hulls") are manually separated. This includes taking a 200 g sample and then manually separating the external fibers that still remain. Subsequently, it is weighed (weight = m). The percentage of residual external fibers is calculated as follows: (m / 200) * 100

[0101] In the method according to the prior art, this percentage is 1.7%. In the method according to the present invention, this percentage is reduced to 0.9%.

[0102] Example 2a: Production of Protein Composition According to the Invention

[0103] According to the improved method of the present invention described above, 75 kg of broad bean powder is prepared. At 20 °C, the powder is suspended in drinking water at 10% by weight of dry matter. The pH value is adjusted to 9 by adding 20% by weight (3.4 kg) of potassium hydroxide. It is still homogenized at 20 °C for 15 minutes. Then the solution is transported to a Sedicanter decanter from Flottweg (rotating drum speed: 60%, i.e., 4657 rpm (about 3500 g), when Vr = 18.8, the screw speed is 60%, pipette the supernatant (overflow) at 140 mm, and the feeding speed is 1 m 3 / h), and the supernatant containing protein is collected.

[0104] The supernatant was acidified to a pH of 4.5 by adding hydrochloric acid (8.2 kg) at a mass percentage of approximately 7%. It was heated to 60 °C by injecting steam into the container jacket and homogenized for 15 minutes here. Again, a Sedicanter decanter from Flottweg (rotating drum speed of 60%, i.e., 4657 rpm (about 3500 g), screw speed of 10% at Vr = 3.5, up to 40% (Vr = 12.6), pipette the overflow at 140 mm until it drops to 137, feed rate of 700 l / h) was used, but this time to recover the precipitate containing the coagulated protein.

[0105] The precipitate was diluted to approximately 15 wt% - 20 wt% dry matter and neutralized to a pH of 7 by adding 20% caustic potash. Heat treatment was carried out at 135 °C using a nozzle, and vacuum flash cooling was implemented at 65 °C. The product was finally atomized (inlet temperature 200 °C, vapor temperature 85 °C - 90 °C).

[0106] The yield of protein extracted from the powder was 86.6%. The resulting protein is called "protein composition according to the present invention".

[0107] Example 2b: Wet Milling Production of Protein Composition According to the Invention

[0108] Small broad bean seeds were first treated with a stone mill Subsequently, the ground material was treated by turbo separation using a system called "serrated" (MZM 1 - 40, ). The air flow rate was 4.0 m.s -1 (23 m 3 .h -1 ). Finally, a light component containing external fibers and a heavy component containing cotyledons were obtained. Subsequently, the heavy component was treated with a knife mill (SM300, ) with a rotational speed of 700 rpm and a 6 mm mesh screen at the outlet. At 20 °C, the heavy component after pre - grinding by the knife mill was suspended in drinking water at 20 wt% dry matter. Subsequently, the heavy component was ground with a Comitrol 19300 mill. The pH was adjusted to 9 by adding approximately 20 wt% caustic potash. It was still homogenized at 20 °C for 15 minutes. Then the solution was fed to a Sedicanter decanter from Flottweg (rotating drum speed: 60%, i.e., 4657 rpm (about 3500 g), at Vr = 18.8, screw rotational speed of 60%, pipette the supernatant (overflow) at 140 mm, feed rate of 1 m 3 / h), and the supernatant containing protein was collected.

[0109] The supernatant was acidified to a pH of 4.5 by adding hydrochloric acid at a mass percentage of approximately 7%. It was heated to 60 °C by injecting steam into the vessel jacket and homogenized for 15 minutes here. The Sedicanter decanter of Flottweg company was used again (the rotary drum speed was 60%, i.e., 4657 rpm (about 3500 g), the screw speed was 10% when Vr = 3.5, up to 40% (Vr = 12.6), and the overflow at 140 mm was pipetted until it dropped to 137, and the feeding speed was 700 l / h), but this time it was to recover the precipitate containing the coagulated protein.

[0110] The precipitate was diluted to about 15 wt% - 20 wt% of dry matter and neutralized to a pH of 7 by adding 20% caustic potash. Heat treatment was carried out at 135 °C using a nozzle, and vacuum flash evaporation cooling was implemented at 65 °C. The product was finally atomized (inlet temperature 200 °C, vapor temperature 85 °C - 90 °C).

[0111] The yield of protein extracted from the powder was 87.8%. The obtained protein was called "protein composition 2b according to the present invention".

[0112] Example 3: Production of Protein Composition According to the Prior Art

[0113] We practiced the teachings of Fernandez - Quintela (Vegetable Foods in Human Nutrition, 51, 1997). First, the small broad bean seeds were processed according to the prior - art method described in paragraph

[0064] . Then the cotyledons were soaked in water for 10 hours and then dried overnight in an incubator at 25 °C. After that, the cotyledons were ground into a powder with an average of 300 microns. It was suspended in drinking water at a water / flour mass ratio of 1 / 5, and the pH of the solution was adjusted to 9.0 with 1N sodium hydroxide. The solution was stirred for 20 min. The insoluble components were separated by centrifugation (4000 g / 20 minutes, 20 °C). The pH of the supernatant was adjusted to 4.0 with 1N hydrochloric acid and stirred for 20 minutes at 20 °C. The solution was centrifuged (4000 g / 20 minutes, 20 °C), and the precipitate was freeze - dried. This protein composition was called: "protein composition of Example 3 according to the prior art"

[0114] Example 4: Function and Analytical Control

[0115] Various compositions obtained by Examples 2 and 3 were compared in terms of analysis (dry matter and protein content) and function (water retention capacity according to Test A and color development L). A commercially available faba bean protein composition, namely 85% faba bean protein isolate (batch number DFC021606181 / C1377) from YANTAI T,FULLBIOTECH CO LTD, was also obtained, which is a representative faba bean isolate available on the market. Table 1 below summarizes these analyses.

[0116] [Table 1]

[0117]

[0118] The table shows the excellent water retention capacity of the protein compositions according to the invention: it is far higher than 3 grams per gram of protein, while the protein compositions according to the prior art hardly exceed 2 grams per gram of protein composition in the best case.

[0119] It can also be noted the excellent protein content, which is higher than 90% in Example 2a.

[0120] The protein content in Example 2b is slightly lower (but still very high compared to isolates of, for example, peas and soybeans), but its water retention capacity is extremely high, three times higher than the protein content in the prior art.

[0121] Example 4: Nutritional Benefits of Protein Composition According to the Invention:

[0122] In this example, the specific nutritional advantages of the protein compositions according to the invention are presented. For this purpose, as a reference, commercially available pea protein compositions, potato protein compositions and milk protein compositions were used as protein compositions of the prior art.

[0123] First, the gastrointestinal digestion of the composition was simulated in vitro, using "Simulated GI digestion of dietary protein: Release of new bioactive peptides involved in gut hormone secretion" (Caron et al., Food Research International, 2016, Vol. 89, Part 1, pp. 382 - 390). The protein was subjected to pepsin hydrolysis (at 37 °C, pH 3, enzyme / protein weight ratio of 1 / 40, for 2 h), followed by trypsin hydrolysis (at 37 °C, pH 7, enzyme / protein weight ratio of 1 / 50, for 2 h). Then, the inhibitory activity of dipeptidyl peptidase - 4 or DPP - IV of the resulting digest was evaluated. DPP - IV is an enzyme involved in cell metabolism, and its inhibition causes a significant increase in the concentrations of glucagon - like peptide - 1 or GLP - 1 (an incretin, i.e., an intestinal hormone, secreted by L cells of the ileum between meals) and glucose - dependent insulinotropic peptide or GIP (a gastro - intestinal hormone, secreted by K cells of the duodenum after meals, which enhances glucose - stimulated insulin secretion in the pancreas). These two hormones lead to an increase in insulin secretion and a decrease in glucagon secretion, a property that can improve the glucose balance in diabetic patients.

[0124] To perform this evaluation, the following protocol was used, which is a modification of the protocol described in "Dipeptidyl peptidase - IV inhibitory activity of dairy protein hydrolysates" (Lacroix and Li - Chan, August 2012, International Dairy Journal 25(2):97 - 102). Briefly, 25 μL of the digest was placed in a test tube, at a concentration range of 1.21 mg.mL -1 to 13.89 mg.mL -1 , in order to react with 75 μL of Tris / HCl buffer (100 mM, pH 8.0) and 25 μL of DPP - IV (0.018 U.mL -1)Pre-incubate for 5 minutes. The reaction was triggered by adding 50 μL of Gly-Pro-p-nitroaniline (1 mM). All samples and reagents were diluted with Tris / HCl buffer. The microplate was incubated at 37 °C for 1 hour, and then the absorbance of the released p-nitroaniline was measured at 405 nm every 2 minutes using a microplate reader (ELx808, Biotek, USA). The DPP-IV inhibition rate was defined as the percentage of DPP-IV activity inhibited by the sample at a given concentration (1 mg.mL -1 )compared to the control result. Then, a graph of the inhibition rate of DPP-IV was established based on the final concentration of the sample. The IC50 (unit: mg / mL) was determined as the final concentration of the sample that inhibited the DPP-IV activity by 50%, in mg / mL. The lower the IC50 value, the better the related inhibitory activity of the sample.

[0125] The results obtained were as follows:

[0126] [Table 2]

[0127]

[0128] The inhibition effect of the faba bean protein composition according to the present invention was good: in fact, its IC50 was half that of the commercially available proteins of the prior art.

[0129] Example 5: So-called "Ready-to-Drink" or RTD Beverage Containing 7% Protein :

[0130] So-called "ready-to-drink" beverages or RTD beverages were prepared to compare the faba bean isolate according to the present invention (2a) and the S85F pea isolate sold by ROQUETTE.

[0131] The formula is shown in Table 3 below:

[0132]

[0133] The preparation method of the beverage is as follows:

[0134] - Mix various powders

[0135] - Heat the water to 50 °C and add the powder mixture

[0136] - Disperse using a Silverson high-shear mixer (30 minutes, 50 °C, 3500 rpm)

[0137] - Heat the oil to 50 °C in another container, add the aqueous dispersion and disperse using a Silverson high-shear mixer (5 minutes, 10000 rpm)

[0138] - Heat treatment at 142 °C for 5 seconds

[0139] High-pressure homogenization at 200 bar for 2 times

[0140] - Cool to 30 °C

[0141] Then, use a Mastersizer 3000 particle size analyzer (Malvern) to measure the particle size by laser diffraction to obtain the particle size distribution line graph of the emulsion in the beverage, and use this to compare various beverages. Use the optical model at 1.50 + 0.01i to directly measure the sample by the liquid method. Measure the coefficients D10, D50, D90, and Dmode well-known to those skilled in the art to characterize the oil emulsion.

[0142]

[0143] By comparing the results, it can be seen that the emulsion obtained using the broad bean isolate according to the present invention is much smaller, which is a sign of a good emulsion.

[0144] Example 6: Plant Milk or "Milk Substitute"

[0145] It is recommended here to use the broad bean isolate 2a according to the present invention to make plant milk.

[0146] The formula is as follows:

[0147]

[0148] The preparation scheme is as follows:

[0149] - Heat the water to 70 °C and hydrate the protein isolate with Sylverson at a speed of 2000 rpm for 15 minutes

[0150] - Add other ingredients except oil and mix for 10 minutes

[0151] - Heat the oil to 65 °C and add the oil with stirring at 6000 rpm

[0152] - UHT sterilization at 142 °C for 5 seconds

[0153] - Homogenize at 75 °C in two stages (270 bar and 30 bar)

[0154] - Cool to 4 °C

[0155] A liquid with the appearance of milk is obtained. This plant milk substitute does not undergo decantation during storage.

[0156] The particle size distribution of the emulsified oil globules was analyzed using a Mastersizer 3000 particle size analyzer (Malvern). The coefficients describing the particle size distribution were as follows: D10 = 0.19 μm, D50 = 0.40 μm, D90 = 0.91 μm. These results are good and fully demonstrate the good emulsification of the lipid globules, just like milk.

[0157] Example 7: Traditional and Light Mayonnaise :

[0158] The good results of isolate 2a according to the present invention in the production of traditional (referred to as "full-fat") and light (referred to as "low-fat") mayonnaise will be demonstrated below.

[0159] The raw materials required to achieve the mayonnaise formulation are as follows:

[0160]

[0161]

[0162] The isolate to be tested will be F85F from ROQUETTE, isolate 2a of broad bean according to the present invention, and aquafaba ("Aquafaba Powder" purchased from the company).

[0163]

[0163] The manufacturing protocol is as follows:

[0164] - In a HOTMIX Pro Gastro, mix the ingredients of the first stage at a speed of 3 for 1 minute (equipment manufacturer: MATFER–FLO, model: 212502).

[0165] - For the low-fat formulation, add the ingredients of the second and third stages at a speed of 4 to 7 within 1 minute and 30 seconds, or for the full-fat formulation, add the ingredient of the second stage at a speed of 3 within 2 minutes.

[0166] - For the full-fat formulation, add the ingredients of the third stage at a speed of 3 within 1 minute.

[0167] - For the full-fat formulation, add the ingredients of the fourth stage at a speed of 3 within 1 minute.

[0168] - Complete emulsification within 1 minute, at a speed of 8 for the low-fat formulation and at a speed of 3 for the full-fat formulation.

[0169] We will use a TA.HDplus texture analyzer (Stable Micro Systems Ltd) to compare the various mayonnaises obtained, so as to measure parameters such as firmness, consistency, and cohesiveness. The firmness (g) corresponds to the force that needs to be applied to press a geometric mold (see the "back extrusion ring" kit described below) into the product. The consistency (g.sec) is the data calculated based on the area under the curve of the firmness. The cohesiveness (g) corresponds to the force that needs to be applied when the geometric mold is removed from the mayonnaise.

[0170] The texture analyzer is equipped with a "back extrusion ring" kit, which consists of 1 disc screwed onto the device and 3 plexiglass containers for filling mayonnaise. The acquisition is completed using Exponent software through a program designed specifically for mayonnaise analysis. The geometric mold drops to the bottom of the container at a speed of 3 mm / s and rises at a speed of 5 mm / s. The software automatically plots the curve over time from which the parameters can be inferred.

[0171] The entire implementation process is clearly explained in the user manual.

[0172] The results of the "low-fat" mayonnaise are as follows:

[0173]

[0174] The results of the "full-fat" mayonnaise are as follows:

[0175]

[0176]

[0177] The results obtained show that the mayonnaise obtained using the broad bean isolate according to the present invention is characterized by good texture values, far higher than those of pea and aquafaba isolates.

[0178] Example 9: Ice Cream :

[0179] Here, pea protein isolate and broad bean isolate according to the present invention in the ice cream formula are compared. pea protein isolate and broad bean isolate according to the present invention.

[0180] The various ice cream compositions are as follows:

[0181]

[0182] The preparation scheme is as follows:

[0183] - Heat the water to 60 °C

[0184] - Add water and 3 / 4 of the sucrose and mix for 5 minutes

[0185] - Add the stabilizer and the remaining sucrose and mix for 5 minutes

[0186] - Add the isolate to be tested and mix for 5 minutes

[0187] - Add coconut oil and mix for 5 minutes

[0188] - Mix in total for 20 minutes at 60 °C

[0189] - High-pressure homogenize at 200 bar at 70 °C

[0190] - At 80 °C, in carry out pasteurization for 3 minutes

[0191] - Cool to 4 °C

[0192] - Place in the refrigerator for overnight ripening

[0193] - Freeze with a Tetrapak freezer, aiming for 100% overrun

[0194] Compare the expansion efficiency of the mixture just obtained before pasteurization. The protocol used is as follows:

[0195] - Pour 1 liter of the mixture into a container

[0196] - Mix at high speed (gear 10) for 6 minutes and then pour into a 2-liter test tube

[0197] - Immediately measure the volume of the mixture and the foam at T0

[0198] - Re-measure after 15 minutes

[0199] The results obtained are as follows:

[0200]

[0201] For the mixture obtained using the isolate according to the present invention, the foam is not visible during the period from the start to 15 minutes. This can be explained as being better retained in the mixture containing the isolate according to the present invention. Such better retained foam can result in ice cream with more uniform expansion.

[0202] Example 10: Gelation at Acidic pH

[0203] Gelling at acidic pH values is an important property, specifically in the production of yogurt or sour cream and tofu.

[0204] After heat treatment and acidification with glucono-delta-lactone (GDL), gel strength control analysis of pea and broad bean isolates was carried out on a TAXT+ texture analyzer.

[0205] The powder was placed in a 60 °C water bath, stirred for 5 minutes, and hydrated in azide water until the dry matter reached 15%. Then the solution was stirred overnight at room temperature. The next day, GDL was added at a weight ratio of 2%. Immediately after the addition, each solution was dispensed into 3 different jars (for three gel strength measurements). Acidification was carried out to bring the pH to 4.6. The samples were placed in an 80 °C water bath for 2 hours and then stored in the refrigerator overnight. The next day, gel strength measurements were performed.

[0206] The gels were characterized at 20 °C using a TAXT+ texture analyzer from Stable Micro Systems Ltd. The parameters were as follows: compression mode, geometric mold: spherical punch P0.5S, pre-test speed: 1 mm / s, test speed: 0.5 mm / s, post-test speed: 10 mm / s, distance: 15 mm, hold time: 60 s, trigger force: 5 g. The force required to apply this displacement was recorded, and the maximum required force was retained.

[0207] The results are as follows:

[0208]

[0209] Obviously, the gel strength obtained using the isolate according to the present invention is 3 times higher than that obtained from pea protein isolate. With this finding, excellent results can be obtained in the manufacture of alternative fermented products for set or drinking yogurts. For set or drinking yogurts, excellent gel strength can enable a formulation without hydrocolloids such as pectin.

[0210] Example 11: Yogurt

[0211] In comparison with the S85F pea protein isolate from Roquette and the broad bean isolate 2a according to the present invention.

[0212] The yogurt formulation is as follows:

[0213]

[0214] The preparation protocol is as follows:

[0215] - At 55 °C, using a Sylverson stirrer, the isolate was hydrated with water at a speed of 2500 rpm for 30 minutes

[0216] - Add other ingredients and mix at a speed of 6000 rpm for 5 minutes

[0217] - High-pressure homogenization at 60 °C in 2 stages (150 bar and 45 bar)

[0218] - Pasteurize at 95 °C for 10 minutes

[0219] - Cool to 42 °C and add YF-L02DA starter culture

[0220] - Maintain at 42 °C for fermentation acidification until a pH of 4.6 is obtained

[0221] - Homogenize at 4000 rpm using an IKA Magic Lab

[0222] - Store at 4 °C for 4 days

[0223] Use a TAXT+ texture analyzer to check the firmness of the resulting yogurt. The results are as follows:

[0224] Yogurt Firmness (g) Pea Raw Material 259 Faba Bean Raw Material 445

[0225] Obviously, the yogurt made from faba bean isolates is firmer because much greater force is required to perform the analysis.

Claims

1. A small broad bean protein composition, characterized in that the color of the small broad bean protein composition has a component L higher than 70 according to the L*a*b measurement method, and the water retention rate according to Test A is higher than 3 grams of water per gram of isolate, wherein the water retention rate according to Test A is obtained as follows: - Weigh 20 g of the sample to be analyzed in a beaker, add drinking water at room temperature until the sample is completely immersed, let it stand in contact for 30 minutes, separate the residual water and the sample with a sieve, and weigh the final weight of the sample after rehydration. - Perform calculations to obtain the water retention capacity = (P - 20) / 20. The small broad bean protein is obtained by the following method, which includes the following steps: 1) Provide small broad bean seeds. 2) Grind the small broad bean seeds using a stone mill, then use an upward air flow to separate the resulting ground material into two components called light and heavy, and then perform secondary grinding on the heavy component using a knife mill. 3) Perform final grinding on the heavy component using a mill selected from a roller mill and a knife mill to obtain a powder, and the average particle size of the powder is between 200 microns and 400 microns. 4) Suspend the powder in an aqueous solvent. 5) Remove the solid components in the suspension by centrifugation and obtain a liquid component. 6) Heat under isoelectric pH conditions and separate the small broad bean protein contained in the liquid component by precipitation, wherein: - The heating temperature is between 45°C and 75°C, and the heating time is between 5 minutes and 25 minutes. - Acidify at a pH value between 4 and 5 to place the liquid component at the isoelectric point. 7) Dilute the previously obtained small broad bean protein to 15 wt% - 20 wt% of the dry matter and neutralize the pH value to between 6 and 8 to obtain the small broad bean protein composition. 8) Dry the small broad bean protein composition.

2. The composition according to claim 1, characterized in that, The component L according to the L*a*b measurement method is higher than 75.

3. The composition according to claim 2, wherein The component L according to the L*a*b measurement method is higher than 80.

4. The composition according to claim 1, wherein The water retention rate according to Test A is higher than 3.5 grams of water per gram of isolate.

5. The protein composition according to claim 1, wherein The protein content of the protein composition expressed as a percentage of protein in the dry matter is higher than 70 wt%.

6. The protein composition according to claim 5, characterized in that, The protein content of the protein composition expressed as a percentage of protein in the dry matter is higher than 80 wt%.

7. The protein composition according to claim 6, wherein The protein content of the protein composition expressed as a percentage of protein in the dry matter is higher than 90 wt%.

8. The protein composition according to claim 1, wherein The dry matter of the protein composition is higher than 80 wt%.

9. The protein composition according to claim 8, wherein The dry matter of the protein composition is higher than 85 wt%.

10. The protein composition according to claim 9, wherein The dry matter of the protein composition is higher than 90 wt%.

11. A method for producing the protein composition according to any one of claims 1 to 10, characterized in that, The method includes the following steps: 1) Provide small broad bean seeds. 2) Grind the small broad bean seeds using a stone mill, then use an upward air flow to separate the resulting ground material into two components called light and heavy, and then perform secondary grinding on the heavy component using a knife mill. 3) Perform final grinding on the heavy component using a mill selected from a roller mill and a knife mill to obtain a powder, and the average particle size of the powder is between 200 microns and 400 microns. 4) Suspend the powder in an aqueous solvent. 5) Remove the solid components in the suspension by centrifugation and obtain a liquid component. 6) Heat at the isoelectric pH and separate the faba bean protein contained in the liquid component by precipitation, where: - The heating temperature is between 45 °C and 75 °C, and the heating time is between 5 minutes and 25 minutes. - Acidify at a pH between 4 and 5 to bring the liquid component to the isoelectric point. 7) Dilute the previously obtained faba bean protein to 15 wt% - 20 wt% of dry matter and neutralize the pH to between 6 and 8 to obtain the faba bean protein composition. 8) Dry the faba bean protein composition.

12. The method according to claim 11, wherein Neutralize the pH to 7 in step 7.

13. The method according to claim 11, characterized in that, The average particle size of the powder obtained in step 3 is 300 microns.

14. The method according to claim 11, wherein Adjust the pH of the aqueous solvent in step 4 to between 8 and 10.

15. The method according to claim 14, wherein Adjust the pH of the aqueous solvent in step 4 to 9.

16. The method according to claim 11, characterized in that Adjust the temperature of the aqueous solvent in step 4 to between 2 °C and 30 °C.

17. The method according to claim 16, characterized in that, Adjust the temperature of the aqueous solvent in step 4 to between 10 °C and 30 °C.

18. The method according to claim 17, characterized in that, Adjust the temperature of the aqueous solvent in step 4 to between 15 °C and 25 °C.

19. The method according to claim 18, wherein Adjust the temperature of the aqueous solvent in step 4 to 20 °C.

20. The method according to claim 11, wherein In step 6, acidify at a pH of 4.

5.

21. The method according to claim 11, wherein The heating temperature is between 50 °C and 70 °C.

22. The method according to claim 21, wherein The heating temperature is between 55 °C and 65 °C.

23. The method according to claim 22, wherein The heating temperature is 60 °C.

24. The method according to claim 11, wherein, The heating time is between 10 and 20 minutes.

25. The method according to claim 24, wherein The heating time is 10 minutes.

26. The method according to claim 11, wherein Step 7 also includes heat treatment.

27. The method according to claim 26, wherein Carry out the heat treatment by directly injecting steam through a nozzle at a temperature of 135 °C and cool by flash evaporation effect under vacuum at 65 °C.

28. The method according to claim 11, wherein Step 8 is dried by multi-effect atomization.

29. The method according to claim 11, wherein Steps 3 and 4 of the method are carried out simultaneously so as to carry out the final grinding on the heavy component in the presence of an aqueous solvent.

30. The method according to claim 29, characterized in that, In the step of carrying out the final grinding on the heavy component in the presence of an aqueous solvent, adjust the pH of the aqueous solvent to between 8 and 10.

31. The method according to claim 30, wherein Adjust the pH of the aqueous solvent to 9.

32. Industrial use of the faba bean protein composition according to any one of claims 1 to 10 or the faba bean protein composition obtained by the method according to any one of claims 11 to 31, specifically its use in human or animal food, cosmetics, and pharmaceuticals.

33. Use according to claim 32, wherein the human or animal food includes: - Beverages, - Yogurt-type fermented milk, - Vegetable cream, - Animal cream, - Ice cream desserts or sorbets, - Biscuits, muffins, pancakes, - Nutrition bars for dietary nutrition, Bread, High-protein cereals, Cheese, Meat analogues, Fish analogues, Sauces.

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