Small broad bean protein composition

Abstract

The present invention relates to the field of legume protein isolates, specifically to faba bean protein isolates, and the solubility of such isolates is less than 25% at a pH value higher than or equal to 7. The present invention also relates to its production method and its industrial applications.

Description

Technical Field

[0001] The present invention relates to the field of legume protein isolates, and more particularly to 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). It belongs to the leguminous plants of the Fabaceae family, Faboideae subfamily, and Fabeae tribe.

[0003] It is of the same species as the broad bean and has been a plant for human consumption 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 topic.

[0006] The traditional method consists of obtaining a bean powder by grinding small broad beans. Then it is diluted in water for alkaline extraction 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 content of the protein isolate thus obtained is at least 80% (expressed as the ratio of the total nitrogen multiplied by the factor 6.25 to the total dry matter, calculated as described in the available document at the following address: http: / / www.favv-afsca.fgov.be / laboratories / methods / fasfc / _documents / METLFSAL003Proteinbrutev10.pdf). This has long been well-known in the industry, especially in human and animal food. In fact, its nutritional and functional properties enable it to be used in many recipes and formulations.

[0008] However, so far, 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, the conventional method of removing the external fibers (referred to in English as "dehulling") does not sufficiently remove the tannins and polyphenols, and the distinct dark color 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 (grading the particles by density using an upward air current). These technical improvements are complex and expensive.

[0012] Secondly, the protein isolate from small broad beans according to the prior art has a high solubility, especially at pH values above 7. This property is essential for some applications, but is disadvantageous for others, such as in the case of bread / pastries.

[0013] Singhal, in his 2015 paper "THE EFFECT OF GENOTYPE AND THE ENVIRONMENT ON THE PHYSICOCHEMICAL AND FUNCTIONAL ATTRIBUTES OF FABA BEAN PROTEIN ISOLATES", very clearly exemplified this high solubility. The process is a classical process that combines alkali extraction and isoelectric precipitation to produce faba bean protein isolate. Subsequently, the author focused on the functional characterization of this isolate, including its solubility. This was measured at pH 7 according to the method used in "Emulsifying properties of chickpea, faba bean, lentil and pea proteins produced by isoelectric precipitation and salt extraction" (Food Research International, Volume 44, 2011, pages 2742 - 2750), and this value is significantly higher than 60%.

[0014] There are multiple solutions, including those proposed in European patent application EP2911524 mentioned above. The solution mainly lies in using lime as a pH regulator. The calcium ion with two positive charges acts as a cross - linker, which links different protein chains together to form a network, and the solubility of this network is lower than that of the same network without lime treatment.

[0015] However, this solution must use lime, and this compound is still difficult to use in an industrial environment. In fact, it is extremely insoluble and is thus processed in the form of an emulsion suspension. Industrial facilities are often blocked by precipitates, which requires shutdown and cleaning.

[0016] Therefore, for this technology, it is important to explore a simple and effective method that can make the faba bean isolate as light - colored as possible and have a low solubility at pH values higher than 7.

[0017] The applicant has the privilege of discovering such a method and such an isolate. 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 which is characterized in that the component L according to the L*a*b measurement method is higher than 70, and the solubility at a pH value higher than 7 is lower than 25%. Preferably, the solubility at a pH value higher than or equal to 7 is lower than 25%. Even more preferably, the solubility at a pH value of 3 is also lower than 25%.

[0019] According to another aspect, a method for producing a small broad bean protein composition according to the present invention is provided, which is characterized in that the method comprises the following steps: 1) providing small broad bean seeds; 2) grinding the small broad bean seeds using a stone mill, and then separating the obtained ground material into two components called light and heavy components by an upward air flow, and then subjecting the heavy component to secondary grinding using a knife mill; 3) finally grinding the heavy component using a roller mill to obtain a powder; 4) suspending the powder in an aqueous solvent with a pH value between 6 and 8, preferably 7; 5) removing the solid components in the suspension by centrifugation and obtaining a liquid component; 6) heating under isoelectric pH conditions and separating the small broad bean protein contained in the liquid component by precipitation; 7) diluting the previously obtained small broad bean protein to 15 wt% - 20 wt% of dry matter and neutralizing the pH value to between 5.5 - 6.5, preferably 6.5, to obtain the small broad bean protein composition; 8) drying the small broad bean protein composition.

[0020] According to the last aspect, an industrial use of the small broad 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 small broad bean protein isolate, the color of which is characterized in that the component L according to the L*a*b measurement method is higher than 70 and the solubility at a pH value higher than 7 is lower than 25%, and its production method and suitable industrial uses are provided.

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

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

[0024] Figure 1

[0025] Figure 1 shows a method for separating the outer fibers and cotyledons of traditional small broad bean seeds;

[0026] Figure 2

[0027] Figure 2 ​​shows a method for separating the outer fibers and cotyledons of small broad bean seeds according to the present invention. Detailed implementation mode

[0028] As described above, according to the present invention, first, a small broad bean protein composition is proposed, and its color is characterized in 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 its solubility at a pH value higher than 7 according to Test A is lower than 25%.

[0029] Preferably, the solubility of the small broad bean protein composition according to the present invention at a pH value higher than or equal to 7 according to Test A is lower than 25% of the total weight.

[0030] Even more preferably, the solubility of the small broad bean protein composition according to the present invention at a pH value higher than or equal to 7 and lower than or equal to 8 according to Test A is lower than 25% of the total weight.

[0031] According to another preferred implementation mode, the solubility of the small broad bean protein composition according to the present invention at a pH value equal to 3 according to Test A is lower than 25% of the total weight.

[0032] "Small broad bean" refers to a leguminous plant belonging to the legume family, Papilionoideae subfamily, 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 through genetic engineering or variety selection are excellent variety sources.

[0033] "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.

[0034] "L*a*b measurement" refers to the color evaluation carried out using an appropriate spectrophotometer according to the colorimetric space method proposed by the CIE (International Commission on Illumination) in the publication "Colorimetry" (2nd edition, 1986, page 36, article 15), which converts the color evaluation into three parameters: the value of brightness L ranges from 0 (black) to 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.

[0035] "Solubility" refers to the percentage of soluble matter in the powder in water, which is quantified by diluting the powder with distilled water, centrifuging the resulting suspension, and analyzing the amount of dissolved matter in the supernatant, and can be measured by the following Test A:

[0036] Add 150 g of distilled water at a temperature of 20 °C ± 2 °C to a 400 mL beaker, stir with a magnetic stir bar, and accurately add 5 g of the bean protein sample to be tested. If necessary, adjust the pH value to the desired value with 0.1 N NaOH. Make up the water to 200 g. Mix at a speed of 1000 rpm for 30 minutes, then centrifuge at 3000 g for 15 minutes. Collect 25 g of the supernatant and place it in a pre-dried and tared crystallizing dish. Place the crystallizing dish in an oven at 103 °C ± 2 °C for 1 hour. Then place it in a desiccator (with dehydrating agent) and cool to room temperature and weigh.

[0037] Solubility corresponds to the content of soluble dry matter, expressed as a percentage of its weight to the weight of the sample. The calculation formula for solubility is as follows:

[0038] [Math.1]

[0039]

[0040] Where:

[0041] P = weight of the sample (unit: g) = 5 g m1 = weight of the crystallizing dish after drying (unit: g)

[0042] m2 = weight of the empty crystallizing dish (unit: g)

[0043] P1 = weight of the collected sample (unit: g) = 25 g

[0044] Preferably, the isolate according to the present invention is characterized in that the protein content expressed as a weight percentage of protein in dry matter is higher than 70%, preferably higher than 80%, and even more preferably higher than 90%.

[0045] Preferably, the weight percentage of dry matter in the protein composition according to the present invention is higher than 80%, preferably the weight percentage is higher than 85%, and even more preferably the weight percentage is higher than 90%. Any method for measuring water content can be used to quantify such dry matter, preferably by a gravimetric technique for evaluating water loss by drying.

[0046] It consists in determining the amount of water evaporated by heating a known quantity of a sample of known mass.

[0047] - First, weigh the sample and measure its mass m1 (unit: g).

[0048] - Place the sample in a heating chamber to evaporate the moisture until the sample mass stabilizes, where the water is completely evaporated. Preferably, the temperature at atmospheric pressure is 105 °C.

[0049] - Weigh the final sample and measure the mass m2 (unit: g).

[0050] - Dry matter = (m2 / m1) * 100.

[0051] The second aspect of the present invention relates to 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) milling the faba bean seeds using a stone mill, then separating the resulting milled material into two components called light and heavy using an upward air flow, and then secondarily milling the heavy component using a knife mill; 3) finally milling the heavy component using a roller mill to obtain a powder; 4) suspending the powder in an aqueous solvent with a pH value between 6 and 8, preferably 7; 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 wt% - 20 wt% of dry matter and neutralizing the pH value to between 5.5 and 6.5, preferably 6.5, to obtain a faba bean protein composition; 8) drying the faba bean protein composition.

[0052] "Stone mill" refers to a system consisting of two superimposed stone columns, where the space left by 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 apply physical stress to these seeds.

[0053] "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.

[0054] The first step consists of providing faba bean seeds. These also include their protective outer fibers, also known as "hulls" in English. The seeds can be subjected to necessary pretreatment steps including cleaning, screening (such as separating the seeds from pebbles), soaking, bleaching, or roasting. Preferably, if bleaching is carried out, the heat treatment conditions are 3 minutes at 80 °C. Non-limiting examples of varieties include, for example, Tiffany, FFS, or YYY varieties. Preferably, faba bean varieties with naturally low tannin and / or polyphenol content will be used, such as the Organdi variety. Such varieties are known and can be obtained through variety hybridization and / or genetic modification.

[0055] The purpose of the second step is to separate the outer fibers and 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, for example, the stone mill sold by the company. As described above, the seeds will be added to the space formed by two stone disks, one of which is rotating. The applicant has noticed that this technique is particularly useful because it will very effectively separate the outer fibers and cotyledons of the seeds. Preferably, the space between the stone disks is adjusted to be between 0.4 and 0.6 mm.

[0056] 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 outer fibers or "hulls" and a "heavy" component mainly containing cotyledons. A particularly suitable specific example of appropriate equipment is, for example, the MZMZ 1-40 sold by the company.

[0057] 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, for example, the SM300 sold by the company.

[0058] The purpose of continuously performing the above three operations in the second step is to very finely separate the outer fibers and 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 fibers or very complex and thus difficult to operate from an industrialization 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 milling operations, two filtering operations, and one turbine separation (using an upward air flow). This method can obtain a cotyledon component that still contains 1.2% of outer fibers. Our invention simplifies this method (using different types of mills for the two milling operations and performing turbine separation between the two milling operations) and can reduce the content of outer fibers to a value of 1% or lower.

[0059] The purpose of the third step is to reduce the particle size of the heavy component rich in cotyledons by milling with a roll mill. A particularly suitable specific example of such a roll mill is, for example, The MLU 202 sold by the company. It is used here to reduce the overall particle size of the powder material, so as to obtain a uniform and sufficiently fine powder material to facilitate the subsequent step 4. The preferred particle size is between 200 and 400 microns, more preferably 300 microns. To measure this particle size, a laser particle size analyzer is preferably used, but any method is feasible, such as sieving.

[0060] Optionally, the step of reducing the particle size of the heavy component rich in cotyledons 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, so they are implemented simultaneously.

[0061] The purpose of the fourth step is to suspend the powder material 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 and salts and sugars. The pH value of the solution is advantageously adjusted to a neutral pH value to maximize the dissolution of tannins and polyphenols. This pH value adjustment can be carried out before and / or after the powder material is suspended in the aqueous solvent.

[0062] 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 6 and 8, preferably 7. Any acidic or alkaline reagent such as soda, lime, citric acid or hydrochloric acid can be considered, but caustic potassium and ascorbic acid are 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.

[0063] The obtained powder material is diluted to obtain a suspension in which the weight of the powder material is between 5% and 25% of the total weight of the water / powder material suspension, preferably between 5% and 15%, preferably between 7% and 13%, 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 and 25 minutes, preferably 10 to 20 minutes, and even more preferably 15 minutes.

[0064] 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.

[0065] The purpose of the sixth step is to acidify the faba bean protein to an isoelectric pH of around 4.5 and then heat the solution to cause the protein called globulin to coagulate, and the globulin is separated by centrifugation.

[0066] The acidification is carried out to a pH 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.

[0067] 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 sugars, salts, and albumin and is called faba bean solubles. It is treated separately, preferably by evaporation and / or drying.

[0068] It should be noted that the prior art for extracting proteins from faba beans only adopts 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 faba 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 faba 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.

[0069] On the other hand, the combination of isoelectric precipitation and controlled heating proposed by the present invention can obtain:

[0070] - Coagulated protein flocs, which after the required treatment result in the product claimed in the present application, and

[0071] - Residual solubles containing other soluble proteins (albumin), salts, sugars, etc.

[0072] The second component can generally be utilized in the fermentation and / or animal nutrition industries. To this end, it needs to be concentrated so as to be stable from a bacteriological perspective. To this end, the conventional operation is to concentrate by vacuum evaporation, which is accomplished using a secondary heating different from that which causes flocculation. During this operation, and in the case of simple isoelectric precipitation during the flocculate / soluble separation process, the coagulated protein deposits will accumulate in the evaporator.

[0073] In the seventh step, the protein composition is subsequently diluted to approximately 15% - 20% by weight of dry matter and neutralized to a pH value between 5.5 and 6.5, preferably 6.5, using any type of alkaline agent, preferably 20% by weight of potassium hydroxide.

[0074] Subsequently, the protein composition can be heat-treated, preferably by directly injecting steam through a nozzle at a temperature of 135 °C and cooling by the flash evaporation effect under a vacuum of 65 °C.

[0075] The resulting protein composition can be used directly, for example, by hydrolysis with proteases or by texturization using an extruder.

[0076] In the eighth step, the protein composition according to the present 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.

[0077] According to a last aspect, an industrial use of the faba bean protein isolate according to the present invention is proposed, specifically its use in human or animal food, cosmetics, and pharmaceuticals. The protein composition according to the present invention can specifically be conveniently used for protein enrichment in bread / pastry applications. For consumers, its high protein content and amino acid profile can achieve beneficial enrichment, and its low solubility can limit aqueous interactions, thereby limiting the interference in the dough or bread dough.

[0078] In human food applications, the protein composition according to the present invention is particularly suitable for dairy product applications.

[0079] More specifically and preferably, the present invention relates to the application of the faba bean isolate in nutritional formulations, such as:

[0080] - Beverages, especially beverages obtained by mixing reconstitutable powders, mainly for dietary nutrition (sports, weight loss), ready-to-drink beverages for dietary or clinical nutrition, clinical nutritional solutions (beverages or enteral bags), plant beverages,

[0081] - Yogurt-type fermented milks (stirred, Greek yogurt, drinking type, etc.),

[0082] - Vegetable cream (such as coffee creamer or "coffee whitener"), dessert cream, ice cream desserts or sorbets,

[0083] - Biscuits, muffins, pancakes, nutritional bars (specifically for weight loss or nutritional products for athletes), bread, especially gluten-free bread rich in protein, high-protein cereals obtained by extrusion cooking (including "potato chips", breakfast cereals, "snacks"),

[0084] - Cheese,

[0085] - Meat analogues, fish analogues, sauces, specifically mayonnaise.

[0086] The nutritional formulation according to the 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 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, for example, the bitterness 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.

[0087] The present invention will be better understood by the following examples.

[0088] Example

[0089] Example 1: Control of traditional conventional external fiber removal method :

[0090] Seeds of the same batch of small broad beans of the Tiffany variety were treated to separate the outer fibers and cotyledons. For this purpose, two methods were employed.

[0091] Methods in the prior art: The seeds are first processed with a knife mill (SM300, ) at a rotational speed of 700 rpm. Subsequently, the milled material is processed by turbo separation using a system called "serrated" (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 ground with a roller mill (MLU 202, ). Finally, a powder with a particle size less than 300 μm is obtained. The method is as Figure 1 shown.

[0092] Improved method according to the present invention: The seeds are first processed with a stone mill . Subsequently, the milled material is processed by turbo separation using a system called "serrated" (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, ) at a rotational speed of 700 rpm and equipped with a 6 mm mesh screen at the outlet. 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 method is as Figure 2 shown.

[0093] For the heavy components obtained by the two methods according to the prior art and the present invention as described above, the residual external fibers (or "hulls") are manually separated. This involves 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

[0094] 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%.

[0095] Example 2: Production of the protein composition according to the present invention

[0096] Using the improved method according to the present invention described above, 75 kg of small broad bean powder was prepared. At 20 °C, the powder was suspended in drinking water at 10% by weight of dry matter. The pH value was adjusted to 7 by adding potassium hydroxide. Homogenization was still carried out at 20 °C for 15 minutes. Then the solution was transferred to a Sedicanter decanter from Flottweg (rotating drum speed: 60%, i.e., 4657 rpm (about 3500 g), when Vr = 18.8, the screw speed was 60%, the supernatant (overflow) at 140 mm was taken with a pipette, and the feeding speed was 1 m 3 / h), and the supernatant containing protein was collected.

[0097] The supernatant was acidified to a pH value of 4.5 by adding hydrochloric acid at about 7% by mass. It was heated to 60 °C by injecting steam into the container jacket and homogenized here for 15 minutes. The Sedicanter decanter from Flottweg was used again (rotating drum speed 60%, i.e., 4657 rpm (about 3500 g), screw speed 10% when Vr = 3.5, up to 40% (Vr = 12.6), the overflow at 140 mm was taken with a pipette until it dropped to 137, and the feeding speed was 700 l / h), but this time it was to recover the precipitate containing coagulated protein.

[0098] The precipitate was diluted to about 15% - 20% by weight of dry matter and neutralized to a pH value of 6.5 by adding 20% potassium hydroxide. Heat treatment was carried out at 135 °C using a nozzle, and vacuum flash evaporation cooling was carried out at 65 °C. The product was finally atomized (inlet temperature 200 °C, vapor temperature 85 - 90 °C).

[0099] The yield of protein extracted from the powder was 72.5%. The obtained protein was called "protein composition according to the present invention".

[0100] Example 3: Production of the protein composition according to the prior art

[0101] This example adopted the literature "Textural properties of legume protein isolate and polysaccharide gels" (Makri et al., Journal of the Science of Food and Agriculture, Vol. 86, 1855 - 1862), which cited the paper "THE EFFECT OF GENOTYPE AND THE ENVIRONMENT ON THE PHYSICOCHEMICAL AND FUNCTIONAL ATTRIBUTES OF FABA BEAN PROTEIN ISOLATES" (Shingha, 2015). Briefly, 350 g - 400 g of powder was dispersed in distilled water (1:10, w / v) and the pH was adjusted to 9.5 with 1M NaOH, then stirred (500 rpm) at 21 - 23 °C for 40 minutes, followed by centrifugation (1600×g, 20 minutes, 4 °C). The supernatant was taken, then diluted with distilled water (1:5, w / v), stirred and centrifuged (1600×g, 20 minutes, 4 °C). The pH of the supernatant was adjusted to 4.5 with 1M HCl, then centrifuged (1600×g, 20 minutes, 4 °C). The supernatant was re - diluted with deionized water, the pH was adjusted to 7.0 with 1M NaOH, and then freeze - dried.

[0102] The yield of protein extracted from the powder was 81.2%. The obtained protein was called "protein composition according to the prior art".

[0103] Example 4: Function and analysis control

[0104] From the perspectives of analysis (dry matter and protein content) and function (solubility according to Test A), various compositions obtained through Examples 2 and 3 were compared. A commercially available faba bean protein composition was also obtained, namely 85% faba bean protein isolate (batch number DFC021606181 / C1377) from YANTAI T, FULL BIOTECH CO LTD, which is a representative faba bean isolate available on the market. Table 1 below summarizes these analyses.

[0105] [Table 1]

[0106]

[0107] This table shows that the protein composition according to the invention has a particularly low solubility at pH values above 7: well below 25%, while the solubility of the protein composition according to the prior art exceeds 35%.

[0108] The gelling ability of different isolates was quantified according to the following protocol:

[0109] 1. An aqueous suspension was prepared by mixing water and the isolate to obtain a final suspension with a dry matter titration concentration of 15% at pH 7;

[0110] 2. The suspension was prepared in a stress-controlled rheometer equipped with a DHR 2 concentric cylinder (TA Instruments);

[0111] 3. The elastic modulus G’ and the viscous modulus G” were measured by applying the following temperature profile:

[0112] a. Stage 1: Heat from 20 °C to 80 °C in 10 minutes

[0113] b. Stage 2: Hold at 80 °C for 110 minutes

[0114] c. Stage 3: Cool from 80 °C to 20 °C in 30 minutes;

[0115] The results were as follows:

[0116]

[0117] It can be seen that the gelling ability is 5 to 6 times higher than that of the isolates of the prior art.

[0118] Example 5: Vegetarian sausage

[0119] The formulation of the broad bean protein isolate according to the invention and the pea isolate sold in a vegetarian formulation as a control was as follows:

[0120]

[0121]

[0122] The protocol for the manufacture of the sausage was as follows:

[0123] - Mix water and crushed ice

[0124] - Disperse methylcellulose into a 60% water / ice mixture using a Kenwood Electronic KM231 (UK). For 5 minutes at maximum speed.

[0125] - Add the protein to be tested and mix using a Kenwood Electronic KM231 (UK). For 10 minutes at maximum speed.

[0126] - Add the oil while stirring at maximum speed and homogenize for an additional 10 minutes.

[0127] - Add the remaining powder ingredients and the remaining 40% water / ice mixture. Stir at maximum speed

[0128] and finally stir for 5 minutes.

[0129] - Stuff into 2-meter-long Viscofan artificial peelable cellulose casings (DAT Schaub, Thiais, France).

[0130]

[0131] - Bake in an industrial oven at 100 °C (Four Bourgeois S2ON1 - serial number S2476057) for 1 hour with the humidity controlled at level 4.

[0132] - Remove the sausages from the oven and allow them to stabilize at room temperature.

[0133] - Manually remove the artificial peelable cellulose casings.

[0134] - Before analysis, boil the sausages in salt-free drinking water for 5 minutes and then leave them at room temperature

[0135] for 30 minutes.

[0136] The resulting sausages will be compared using a TAXT2i rheometer (Stable Micro Systems, Texture Analyzer Model XT2i, UK) and its software version 2.64.

[0137] A test called "slice" or "cut" will be performed to characterize the sausages, which consists of using a texture analyzer to measure the necessary force to perform the action of dividing the sausage into two parts. This test is carried out using a Warner - Bratzler shear, which penetrates the sausage completely by 25 mm with a minimum detection limit of 0.06 N. Characterization is carried out using the maximum force at the break point.

[0138] The resulting values are as follows:

[0139]

[0140] It can be seen that the force required to slice sausage 3 is much higher than the force required to slice the sausages obtained from commercially available pea isolates. From this result, the conclusion can be drawn that the sausages obtained using the broad bean isolate according to the present invention are firmer.

[0141] Example 6: Traditional and light mayonnaise

[0142] ​The good results of the isolates according to the invention in the production of traditional (referred to as "full-fat") and light (referred to as "low-fat") mayonnaise will be demonstrated below.

[0143] The raw materials required to achieve the mayonnaise formulation are as follows:

[0144]

[0145]

[0146] The isolates to be tested will be Roggli's F85F, the broad bean isolate according to the invention, and aquafaba ("Aquafaba Powder" purchased from the company).

[0147] The manufacturing protocol is as follows:

[0148] - In the HOTMIX Pro Gastro, mix the ingredients of Phase 1 at speed 3 for 1 minute (equipment manufacturer: MATFER–FLO, model: 212502).

[0149] - For the low-fat formulation, add the ingredients of Phases 2 and 3 at speeds 4 to 7 within 1 minute 30 seconds, or for the full-fat formulation, add the ingredients of Phase 2 at speed 3 within 2 minutes.

[0150] - For the full-fat formulation, add the ingredients of Phase 3 at speed 3 within 1 minute.

[0151] - For the full-fat formulation, add the ingredients of Phase 4 at speed 3 within 1 minute.

[0152] - Complete emulsification within 1 minute, at speed 8 for the low-fat formulation and at speed 3 for the full-fat formulation.

[0153] We will use a TA.HDplus texture analyzer (as shown in Appendix 1) to compare the various mayonnaises obtained, which allows measurement of parameters such as firmness, consistency, and cohesiveness. Firmness (g) corresponds to the force required to press a geometric mold (see the "back extrusion ring" kit described below) into the product, consistency (g.sec) is calculated from the area under the firmness curve, and cohesiveness (g) corresponds to the force required to remove the geometric mold from the mayonnaise.

[0154] The texture analyzer is equipped with a "back extrusion loop" kit, which consists of 1 disc screwed onto the device and 3 plexiglass containers for filling mayonnaise. The acquisition is done 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 parameters can be inferred.

[0155] The entire implementation process is clearly explained in the user manual.

[0156] The results for "low-fat" mayonnaise are as follows:

[0157]

[0158] The results for "full-fat" mayonnaise are as follows:

[0159]

[0160] The results obtained show that the mayonnaise obtained with the broad bean isolate according to the invention is characterized by good texture values, which are higher in the "low-fat" mayonnaise than those of the pea and aquafaba isolates.

[0161] Example 7: Plant milk or "milk substitute"

[0162] Plant milk is prepared to evaluate the performance of our isolate according to the invention in this application.

[0163] The formula is as follows:

[0164]

[0165]

[0166] The preparation protocol is as follows:

[0167] - Heat the water to 70 °C and hydrate the protein isolate with Sylverson at 2000 rpm for 15 minutes

[0168] - Add the other ingredients except oil and mix for 10 minutes

[0169] - Heat the oil to 65 °C and add the oil with stirring at 6000 rpm

[0170] - UHT sterilize at 142 °C for 5 seconds

[0171] - Homogenize at 75 °C in 2 stages (270 bar and 30 bar)

[0172] - Cool to 4 °C

[0173] The particle size distribution of the emulsified oil globules was analyzed using a Mastersizer particle size analyzer. The particle size measurement parameters were as follows: D10 = 0.21 μm, D50 = 0.45 μm, D90 = 1.42 μm.

[0174] These results are good and fully demonstrate the good emulsification of the lipid globules, just like in milk.

Claims

1. A small broad bean protein composition, characterized in terms of the color of the small broad bean protein composition, wherein the component L according to the L*a*b measurement method is higher than 70, and the solubility at a minimum pH of 7 and less than or equal to 8 according to Test A is less than 25% of the total weight. Test A is defined as follows: - Add 150 g of distilled water at a temperature of 20 °C ± 2 °C to a 400 mL beaker, stir with a magnetic stir bar, and accurately add 5 g of the bean protein sample to be tested; - If necessary, adjust the pH to the desired value with 0.1 N NaOH; - Make up the water to 200 g; - Mix at a speed of 1000 rpm for 30 minutes, and then centrifuge at 3000 g for 15 minutes; - Collect 25 g of the supernatant and place it in a pre-dried and tared crystallizing dish; - Place the crystallizing dish in an incubator at 103 °C ± 2 °C for 1 hour; - Then place it in a desiccator with a dehydrating agent and cool it to room temperature and weigh it; Wherein, The solubility corresponds to the content of soluble dry matter, expressed as a percentage of its weight to the weight of the sample. The calculation formula for solubility is as follows: Wherein, P is the weight of the sample, unit: g; m1 is the weight of the crystallizing dish after drying, unit: g; m2 is the weight of the empty crystallizing dish, unit: g; P1 is the weight of the sample collected, unit: g, The small broad bean composition 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, and then use an upward air flow to separate the obtained ground material into two components called light and heavy, and then perform secondary grinding on the heavy part using a knife mill; 3) Perform final grinding on the heavy component using a roller mill to obtain a powder, and the average particle size of the powder is between 200 and 400 microns; 4) Suspend the powder in an aqueous solvent with a pH between 6 and 8; 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 acidification of the liquid component is carried out at a pH between 4 and 5, - The heating temperature is between 45 °C and 75 °C, and the heating time in step 6 is between 5 minutes and 25 minutes; 7) Dilute the small broad bean protein obtained previously to a dry matter weight content of 15 wt% - 20 wt%, and neutralize it to a pH between 5.5 and 6.5, To obtain the small broad bean protein composition; 8) Dry the small broad bean protein composition.

2. The protein composition according to claim 1, wherein The small broad bean protein composition is characterized in terms of the color, wherein the component L according to the L*a*b measurement method is higher than 75.

3. The protein composition according to claim 2, wherein The small broad bean protein composition is characterized in terms of the color, wherein the component L according to the L*a*b measurement method is higher than 80.

4. The protein composition according to claim 1, wherein The solubility at a pH equal to 3 according to Test A is less than 25% of the total weight.

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

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

7. The protein composition according to claim 6, wherein The protein composition has a protein weight content that is 90% by weight higher than the protein percentage of the protein in the dry matter.

8. The protein composition according to any one of claims 1 to 7, characterized in that, The dry matter weight percentage of the protein composition is higher than 80% by weight.

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

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

11. A method for producing the protein composition according to any one of claims 1 to 10, characterized in that, The method comprises the following steps: 1) Providing small broad bean seeds; 2) Grinding the small broad bean seeds using a stone mill, then separating the resulting grind into two components called light and heavy using an upward air current, and then secondarily grinding the heavy component using a knife mill; 3) Finally grinding the heavy component using a roller mill to obtain a powder, the average particle size of the powder being between 200 and 400 microns; 4) Suspending the powder in an aqueous solvent with a pH between 6 and 8; 5) Removing the solid components from the suspension by centrifugation and obtaining a liquid component; 6) Heating under isoelectric pH conditions and separating the broad bean protein contained in the liquid component by precipitation, wherein: - The acidification of the liquid component is carried out at a pH between 4 and 5, - The heating temperature is between 45°C and 75°C, and the heating time in step 6 is between 5 minutes and 25 minutes; 7) Diluting the broad bean protein obtained previously to a dry matter weight content of 15% - 20% and neutralizing to a pH between 5.5 and 6.5, to obtain the broad bean protein composition; 8) Drying the broad bean protein composition.

12. The production method of the protein composition according to claim 11, characterized in that, In step 4, the pH of the aqueous solvent is 7.

13. The method for producing the protein composition according to claim 11 or 12, characterized in that, In step 7, the pH is neutralized to 6.

5.

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

15. The method according to claim 11, wherein In step 4, the temperature of the aqueous solvent is adjusted to between 2°C and 30°C.

16. The method according to claim 15, characterized in that, In step 4, the temperature of the aqueous solvent is adjusted to between 10°C and 30°C.

17. The method according to claim 16, wherein In step 4, the temperature of the aqueous solvent is adjusted to between 15°C and 25°C.

18. The method according to claim 17, wherein In step 4, the temperature of the aqueous solvent is adjusted to 20°C.

19. The method according to claim 11, characterized in that, The acidification of the liquid component in step 6 is carried out at a pH of 4.

5.

20. The method according to claim 11, wherein, In step 6, the pH of the liquid component is adjusted using ascorbic acid.

21. The method according to claim 11, wherein The heating temperature in step 6 is between 50°C and 70°C.

22. The method according to claim 21, wherein The heating temperature in step 6 is between 55°C and 65°C.

23. The method according to claim 22, wherein The heating temperature in step 6 is 60°C.

24. The method according to claim 11, wherein The heating time in step 6 is between 10 and 20 minutes.

25. The method according to claim 24, wherein The heating time in step 6 is 10 minutes.

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

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

28. Industrial use of the broad bean protein composition obtained by the method according to any one of claims 1 to 10 or according to any one of claims 11 to 27.

29. The industrial use of the small broad bean protein composition according to claim 28, characterized in that, Specifically for use in human or animal food, cosmetics, and pharmaceuticals.

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