阅读说明：本技术 小蚕豆蛋白组合物 (Vicia faba protein composition ) 是由 J·L·文图莱拉 D·帕斯 C·拉罗切 于 2020-03-25 设计创作，主要内容包括：本发明涉及豆类蛋白分离物领域,具体涉及小蚕豆蛋白分离物,并且这种分离物在pH值高于或等于7时的溶解度低于25％。本发明还涉及其生产方法及其工业应用。(The present invention relates to the field of legume protein isolates, and in particular to fava bean protein isolates and which have a solubility of less than 25% at a pH of greater than or equal to 7. The invention also relates to a method for the production thereof and to the industrial use thereof.)

1. A fava bean protein composition, said fava bean protein composition being characterised by a colour wherein the component L, measured according to la b, is higher than 70, preferably higher than 75, even more preferably higher than 80, and the solubility according to test a at a pH value higher than or equal to 7 is lower than 25% of the total weight.

2. The protein composition according to claim 1, wherein the solubility according to test a is lower than 25% of the total weight at a pH value higher than or equal to 7 and lower than or equal to 8.

3. The protein composition according to any one of claims 1 to 2, characterized in that the solubility according to test A at a pH value equal to 3 is lower than 25% of the total weight.

4. Protein composition according to any one of claims 1 to 3, characterized in that it has a protein content higher than 70% by weight, preferably higher than 80% by weight, even more preferably higher than 90% by weight, expressed as percentage of protein in dry matter of protein.

5. The protein composition according to any one of claims 1 to 4, characterized in that the weight percentage of dry matter in the protein composition is higher than 80 weight%, preferably the weight percentage is higher than 85 weight%, even more preferably the weight percentage is higher than 90 weight%.

6. A method for producing a protein composition according to any one of claims 1 to 5, characterized in that said method comprises the following steps:

1) providing small broad bean seeds;

2) milling the seeds of fava beans using a stone mill, then separating the resulting mill into two components called light and heavy using an ascending gas stream, then secondary milling the heavy fraction with a knife mill;

3) final milling of the heavy fraction using a roller mill to obtain a meal;

4) suspending the powder in an aqueous solvent having a pH between 6 and 8, preferably 7;

5) removing solid components from the suspension by centrifugation and obtaining liquid components;

6) heating under isoelectric pH, and separating small broad bean protein contained in liquid component by precipitation;

7) diluting the fava bean protein obtained previously to a content of 15% to 20% by weight of dry matter content by weight and neutralizing to a pH value comprised between 5.5 and 6.5, preferably 6.5, to obtain said fava bean protein composition;

8) drying the fava bean protein composition.

7. Process according to claim 6, characterized in that the average particle size of the powder obtained in step 3 is between 200 and 400 microns, preferably 300 microns.

8. The method according to any one of claims 6 to 7, characterized in that the temperature of the aqueous solvent in step 4 is adjusted to be between 2 ℃ and 30 ℃, preferably between 10 ℃ and 30 ℃, preferably between 15 ℃ and 25 ℃, even more preferably 20 ℃.

9. The method according to any one of claims 6 to 8, wherein the acidification of the liquid component in step 6 is carried out at a pH between 4 and 5, preferably 4.5.

10. The method according to any one of claims 6 to 9, wherein the pH of the liquid component is adjusted in step 6 with ascorbic acid.

11. The method according to any one of claims 6 to 10, wherein the heating temperature in step 6 is between 45 ℃ and 75 ℃, preferably between 50 ℃ and 70 ℃, even more preferably between 55 ℃ and 65 ℃, most preferably 60 ℃, and the heating time in step 6 is between 5 minutes and 25 minutes, preferably between 10 and 20 minutes, most preferably 10 minutes.

12. The method according to any one of claims 6 to 11, wherein step 7 further comprises a heat treatment, preferably direct injection of steam through a nozzle at a temperature of 135 ℃ and cooling by flash effect under vacuum at 65 ℃.

13. Industrial use of the fava bean protein composition according to any one of claims 1 to 5 or obtained by the method according to any one of claims 6 to 12, in particular in human or animal food, cosmetics, pharmaceuticals.

Technical Field

The invention relates to the field of bean protein isolate, in particular to a small broad bean protein isolate.

Background

Fava beans (fava or faverole) are annual plants of the fava bean species (Vicia faba). Leguminous plants belonging to the family Leguminosae (Fabaceae), subfamily Papilionaceae (Faboideae), and family Vicia (Fabae).

It is the same species as broad bean, a plant for human consumption since ancient times. Thus, the term broad bean refers to both seeds and plants.

Various production methods are known in the art, which can utilize the seeds of fava beans to produce protein isolates.

The current knowledge of this subject was reviewed in the book of broad of beans as a future protein supply in the human diet (Multar et al, Comprehensive RevieWS in Food Science and Food Safety, 2015, 14).

The traditional method consists in obtaining bean flour by milling fava beans. It is then diluted in water for alkaline extraction with the aim of dissolving the fava bean proteins. The solution is then subjected to liquid/solid separation, so as to obtain, on the one hand, a crude protein solution and, on the other hand, a solid fraction enriched in starch and fibres. The protein is extracted by isoelectric pH precipitation of the protein, separated from the aqueous solution and dried.

The protein isolate thus obtained has a protein content of at least 80% (expressed as the total nitrogen multiplied by the ratio of the factor 6.25 to the total dry matter, calculated as described in the available documents at http:// www.favv-afsca. fgov. be/laboratories/methods/fasffc/_ documents/METLFSAL 003Proteinebortev 10. pdf). This has long been known in industrial terms, particularly in the context of human and animal food. In fact, its nutritional and functional properties make it useful in a wide variety of recipes and formulas.

However, up to now, the skilled person has to cope with two major technical problems.

First, the resulting protein isolate is typically characterized as appearing dark gray, or even black. This is mainly due to the presence of tannins and polyphenols in the outer fibres produced by the proteins during the manufacture of the protein isolate.

Despite great care, the traditional external fiber removal method (known as "dewling") does not sufficiently remove tannins and polyphenols and presents a distinctly dark color limiting the range of possible applications.

Optimized methods have been developed. For example, the method described in technical-scale shelling process to improve the nutritional value of faba beans (Meijer et al, Animal Feed Science and Technology, 1994, 46) proposes two mills, two filters and one turbine separation (particle fractionation by density using an updraft). These techniques are complex and expensive to develop.

Secondly, the small broad bean protein isolates according to the prior art have a high solubility, especially at pH values above 7. This property is essential for some applications but disadvantageous for other applications such as bread/pastry.

Singhal, in its 2015 paper "influence OF GENOTYPE AND ENVIRONMENT ON PHYSICOCHEMICAL AND FUNCTIONAL properties OF fava BEAN PROTEIN isolate (THE EFFECT OF GENOTTE AND THE ENVIRONMENT ON THE PHOBICOCHEMICAL AND FUNCTION OF FABA BEAN PROTEIN ISOLATES)" very clearly illustrates this high solubility. The process is a classical process for producing the isolated protein of fava beans by combining alkali extraction and isoelectric precipitation. Subsequently, the authors focused on the functional characterization of this isolate, including its solubility. This is measured at a pH of 7, according to the method used in "emulsification Properties of chickpea, Vicia faba, lentil and pea proteins produced by isoelectric precipitation and salt extraction" (emulsion properties of chickpea, faba bean, lens and pea proteins produced by the inventive method by iso-electric precipitation and salt extraction), which is clearly higher than 60%.

There are various solutions, including those proposed in the above european patent application EP 2911524. The proposal mainly uses lime as a pH value regulator. Calcium ions with two positive charges act as cross-linking agents that link different protein chains together, thereby forming a lattice whose solubility is lower than that of the same lattice that has not been limed.

However, this solution necessitates the use of lime, and the use of such compounds in industrial environments is still difficult. In fact, it is very insoluble and therefore handled in the form of a milky suspension. Industrial facilities are often clogged with deposits, which require shut-down and cleaning.

Therefore, it is important to find a simple and effective method for making the small broad bean isolate as light-colored as possible and less soluble at pH values above 7.

Applicants have fortunately discovered such methods and such isolates. Hereinafter, the present invention will be described.

Disclosure of Invention

As mentioned above, according to the present invention, firstly a fava bean protein composition is proposed, the colour of which is characterized by a component L higher than 70, preferably higher than 75, even more preferably higher than 80, measured according to la ab, and a solubility at pH values higher than 7 lower than 25%, according to test a.

Preferably, the fava bean protein composition according to the invention has a solubility according to test a at a pH value higher than or equal to 7 lower than 25% of the total weight.

Even more preferably, the fava bean protein composition according to the invention has a solubility according to test a at a pH value higher than or equal to 7 and lower than or equal to 8 lower than 25% of the total weight.

According to another preferred embodiment, the small broad bean protein composition according to the invention has a solubility according to test a at a pH value equal to 3 lower than 25% of the total weight.

The "fava beans" are leguminous plants belonging to the family leguminosae, the subfamily Papilionaceae, and the family Vicia victoriae, and are annual plants of the species Vicia faba. People divide it into "minor" and "major" varieties. In the present invention, both a wild variety and a variety obtained by genetic engineering or variety selection are excellent sources of varieties.

By "protein composition" is meant any protein-rich composition obtained by plant extraction, and if necessary by purification. Concentrates with a protein content higher than 50% of dry matter are distinguished from isolates with a protein content higher than 80% of dry matter.

"L a b measurement" means a colorimetric evaluation performed by an appropriate spectrophotometer according to the colorimetric space method proposed by CIE (international commission on illumination) in the publication "Colorimetry" (1986, 2 nd edition, page 36, bar 15), which converts the colorimetric evaluation into 3 parameters: the value of the luminance L is between 0 (black) and 100 (white reference); the parameter a represents the value on the green → red axis, and the parameter b represents the value on the blue → yellow axis. Such COLOR measurement is preferably carried out using a DATA COLOR-DATA FLASH 100 or KONIKA MINOLTA CM5 spectrophotometer with the aid of its user manual.

"solubility" means the percentage of solubles in water of a powder quantified by diluting the powder with distilled water, centrifuging the resulting suspension and analyzing the amount of dissolved material in the supernatant, as measured by test A below:

150g of distilled water at 20 ℃ +/-2 ℃ are added to a 400mL beaker while stirring with a magnetic bar and 5g of the pulse protein sample to be tested are added precisely. If necessary, the pH is adjusted to the desired value with 0.1N NaOH. Make up water to 200 g. Mix at 1000rpm for 30 minutes and then centrifuge at 3000g for 15 minutes. 25g of the supernatant was collected and placed in a crystallization dish which had been previously dried and de-skinned. The dish was placed in an incubator at 103 ℃ +/-2 ℃ for 1 hour. It was then placed in a desiccator (with dehydrating agent) to cool to room temperature and weighed.

Solubility corresponds to the content of soluble dry matter, expressed as a percentage of its weight on the weight of the sample. The solubility is calculated as follows:

[Math.1]

wherein:

weight of sample (unit: g) 5g

m1 weight of dried crystallizing dish (unit: g)

m2 weight of empty petri dish (unit: g)

Weight of collected sample (unit: g) 25g

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

Preferably, the weight percentage of dry matter in the protein composition according to the invention is higher than 80%, preferably the weight percentage is higher than 85%, even more preferably the weight percentage is higher than 90%. Any method for measuring the water content can be used for quantifying this dry matter, preferably a gravimetric technique for assessing water loss by drying.

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

First, the sample is weighed and the mass m1 (units: g) is measured.

-placing the sample in a heating chamber to evaporate water until the sample mass stabilizes, wherein the water is completely evaporated. Preferably, the temperature at atmospheric pressure is 105 ℃.

The final sample was weighed and the mass m2 (units: g) was measured.

Dry matter (m2/m1) × 100.

A second aspect of the present invention consists in a method for producing a fava bean protein composition according to the invention, characterized in that it comprises the following steps: 1) providing small broad bean seeds; 2) milling seeds of fava beans using a stone mill, separating the resulting mill into two components called light and heavy using an ascending gas stream, and then secondary milling the heavy component using a knife mill; 3) final milling of the heavy fraction using a roller mill to obtain a meal; 4) suspending the powder in an aqueous solvent having a pH between 6 and 8, preferably 7; 5) removing solid components from the suspension by centrifugation and obtaining liquid components; 6) heating under isoelectric pH, and separating small broad bean protein contained in liquid component by precipitation; 7) diluting the previously obtained fava bean proteins to 15-20 wt% of dry matter and neutralizing the pH to between 5.5 and 6.5, preferably 6.5, to obtain a fava bean protein composition; 8) drying the fava bean protein composition.

"Stone mill" means a system consisting of two stacked stone columns, where the space left by the stone columns is approximately equal to the seed size. One of the cylinders is stationary and the other cylinder is rotating. Seeds are introduced between the two cylinders and their relative movement will exert a physical stress on the seeds.

"knife grinder" means a system consisting of a chamber equipped with an upper inlet for the addition of seeds, a plurality of knives arranged on a rotating shaft in said chamber, and a lower outlet for the outflow of only the seeds of the desired size.

The first step consists in providing seeds of fava beans. These also include their protective outer fibers, also known as "hells" in english. The seeds may be subjected to the necessary pre-treatments including cleaning, sieving (e.g. to separate the seeds from the pebbles), soaking, bleaching or baking steps. Preferably, if bleaching is performed, the heat treatment conditions are 80 ℃ for 3 minutes. Non-limiting examples of varieties include, for example, Tiffany, FFS, or YYY varieties. Preferably, a small broad bean variety, such as the Organdi variety, will be employed which is naturally lower in tannin and/or polyphenol content. Such varieties are known and can be obtained by variety crossing and/or genetic modification.

The objective of the second step is to separate the outer fibres and the cotyledons as efficiently as possible. Firstly, milling seeds of the small broad beans by a stone mill for one time. A particularly suitable specific example of such a stone mill is, for exampleA stone mill sold by the company. As mentioned above, the seed will be added to the space formed by the two stones, one of which is rotating. The applicant has noted that this technique is particularly useful because it allows a very efficient separation of the outer fibres of the seed from the cotyledons. Preferably, the space between the stone discs is adjusted to be between 0.4 and 0.6 mm.

The resulting millbase is then subjected to a counter-current ascending gas flow. Different solid particles will be classified according to their density. Generally, two components are obtained after stabilization: a light component containing mainly external fibers or "hells" and a "heavy" component containing mainly cotyledons. Particularly suitable specific examples of suitable devices are for exampleMZMZ 1-40 sold by the company.

The heavy cotyledon-rich component will then be milled using a knife mill. A particularly suitable specific example of such a stone mill is, for exampleSM300 sold by companies.

The purpose of performing the above three operations in succession in the second step is to separate the outer fibres and the cotyledons very finely, while avoiding damage and mixing of these two parts. The prior art methods are either too simple to effectively separate the outer fibers or are complicated and therefore difficult to operate from an industrial point of view. For example, the method described in technical-scale shelling process to improve the nutritional value of faba beans (Meijer et al, Animal Feed Science and Technology, 46, 1994) proposes two mills, two filters and one turbine separation (using an updraft). The method can obtain cotyledon component, and the cotyledon still contains 1.2% of external fiber. Our invention simplifies the process (two mills with different technology mill types, turbo separation between the two mills) and can reduce the content of external fibres to values of 1% or less.

The purpose of the third step is to reduce the particle size of the cotyledon-rich heavy fraction by roller milling. A particularly suitable specific example of such a roller mill is, for exampleMLU 202 sold by the company. It is used here to reduce the overall particle size of the powder, so that a homogeneous and sufficiently fine powder is obtained, which facilitates the subsequent step 4. Preferably the particle size is between 200 and 400 microns, more preferably 300 microns. For measuring the particle size, a laser particle sizer is preferably used, but any method is feasible, such as sieving.

Alternatively, the step of reducing the particle size of the cotyledon-rich heavies may be carried out in the presence of an aqueous solvent, preferably water. In this case, the following fourth step is merged with the third step, and thus they are performed simultaneously.

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 aim is to perform a selective extraction of certain compounds, mainly proteins, as well as salts and sugars, by dissolving them. The pH of the solution is advantageously adjusted to a neutral pH to maximize tannin and polyphenol dissolution. This pH adjustment may be performed before and/or after the powder is suspended in the aqueous solvent.

The aqueous solvent is preferably water. However, a compound that can promote dissolution, for example, may be added to the aqueous solvent. The pH of the aqueous solvent is adjusted to between 6 and 8, preferably 7. Any acidic or alkaline agent such as soda, lime, citric acid or hydrochloric acid may be considered, but caustic potash and ascorbic acid are preferred. The temperature is adjusted to between 2 ℃ and 30 ℃, preferably between 10 ℃ and 30 ℃, preferably between 15 ℃ and 25 ℃, even more preferably 20 ℃. This temperature needs to be adjusted throughout the extraction reaction.

The powder obtained is diluted so as to obtain a suspension of between 5% and 25% by weight of powder, preferably between 5% and 15%, preferably between 7% and 13%, even more preferably between 9% and 11%, most preferably 10%, of the total weight of the water/powder suspension. The suspension is stirred by any means known to those skilled in the art, such as a vessel equipped with a stirrer, with blades, with a marine blade or with any effective fermentation device. The extraction time, preferably with simultaneous stirring, is between 5 and 25 minutes, preferably 10 to 20 minutes, even more preferably 15 minutes.

The purpose of the fifth step is to centrifuge the soluble or solid fraction obtained in the fourth step. Preferred industrial principles can be found in european patent application EP1400537, which is incorporated herein by reference. The principle of the process is to first extract the starch rich fraction using a cyclone separator and then extract the internal fiber rich fraction using a horizontal decanter. However, industrial centrifuges may also be used to extract components rich in starch and internal fibers. In any case a solid fraction and a liquid fraction concentrated in the major part of the protein are obtained.

The purpose of the sixth step is to acidify the fava bean proteins to an isoelectric pH of around 4.5 and then heat the solution to coagulate the proteins called globulins, which are centrifuged.

Acidifying to pH 4-5, preferably 4.5. Preferably, this is carried out using about 7% by mass of hydrochloric acid, but any type of acid, mineral or organic, such as citric acid, can be used. Even more preferably, it is possible to use pure ascorbic acid or in combination with other mineral or organic acids. Acidification with ascorbic acid can improve the final color development. Any subsequent heating means is possible, for example by means of a stirred tank equipped with a jacket and/or coils or an in-line steam jet cooker (jet cooker). The heating temperature is advantageously between 45 ℃ and 75 ℃, preferably between 50 ℃ and 70 ℃, even more preferably between 55 ℃ and 65 ℃, most preferably 60 ℃. The heating time is advantageously between 5 minutes and 25 minutes, preferably between 10 and 20 minutes, most preferably 10 minutes.

Protein compositions whose major component is globulin will coagulate and precipitate in solution. It can be separated by any centrifugation technique, e.g.Sedicanteur. The resulting residual solution is concentrated in sugar, salt and albumin and is called fava bean solubles. It is treated separately, preferably by evaporation and/or drying.

It should be noted that the prior art for extracting proteins from fava beans only adopts isoelectric precipitation without heating. By combining the two steps according to the invention, it is possible to obtain the isolate according to the invention, but also to obtain the temperature-stable fava bean solubles (name of the supernatant obtained after precipitation and centrifugation). In fact, when the soluble fraction of fava beans obtained by isoelectric precipitation is exposed to high temperatures, for example in an evaporator, precipitation occurs. This precipitate is a major disadvantage as it can lead to blockage of industrial facilities.

On the other hand, the isoelectric precipitation provided by the invention is combined with controlled heating, so that the following can be obtained:

coagulated protein flocs, which after the desired treatment result in the products claimed in this application, and

residual solubles containing other soluble proteins (albumin), salts and sugars etc.

The second ingredient may typically be utilized in the fermentation and/or animal nutrition industries. For this purpose, it needs to be concentrated in order to be stable from a bacteriological point of view. For this reason, the conventional practice is to concentrate by vacuum evaporation, which is done by means of a second heating different from that which would coagulate the flocs. In this operation, and in the case of simple isoelectric precipitation during the floc/solubles separation, the coagulated protein deposit will accumulate in the evaporator.

In the seventh step, the protein composition is then diluted to about 15% to 20% by weight of dry matter and neutralized to a pH between 5.5 and 6.5, preferably 6.5, using any type of alkaline agent, preferably 20% by weight of caustic potash.

The protein composition may then be subjected to a heat treatment, preferably by direct injection of steam through a nozzle at a temperature of 135 ℃ and cooling by a flash effect under vacuum at 65 ℃.

The resulting protein composition can be used directly, for example by hydrolysis with a protease or texturization by means of an extruder.

In an eighth step, the protein composition according to the invention is dried. The preferred drying means is atomization, particularly with a multi-effect atomizer. Typical parameters are an inlet temperature of 200 ℃ and a vapor temperature of 85-90 ℃.

According to a final aspect, the industrial use of the fava bean protein isolate according to the invention is proposed, in particular in human or animal food, cosmetics, pharmaceuticals. The protein composition according to the invention is particularly convenient for use in protein enrichment in bread/pastry applications. For the consumer, a high protein content and its amino acid profile may enable a beneficial enrichment, and its low solubility may limit aqueous interactions and thus interference of the dough or bread dough.

In human food applications, the protein composition according to the invention is particularly suitable for dairy applications.

More particularly and preferably, the present invention relates to the use of fava bean isolates in nutritional formulations, such as:

beverages, in particular beverages obtained by mixing powders to be reconstituted, mainly for dietetic nutrition (sports, weight loss), ready-to-drink beverages for dietetic or clinical nutrition, clinical nutritional liquids (beverages or enteral bags), vegetable beverages,

yogurt type fermented milk (stirred type, greek yogurt, drinkable type, etc.),

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

biscuits, muffins, pancakes, nutritional bars (for slimming or nutrition for athletes specifically), bread, in particular gluten-free bread rich in proteins, high-protein cereals obtained by extrusion cooking (including "potato chips", breakfast cereals, "snacks"),

-a cheese, the cheese being,

-meat analogue, fish analogue, sauce, in particular mayonnaise.

The nutritional formula according to the invention may also comprise other ingredients which may alter the chemical, physical, organoleptic or processing properties of the product or act as a pharmacotrophic or supplemental ingredient for certain target populations. Many of these optional ingredients are known or otherwise used in other food products and may also be used in the nutritional formulas 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 selected products. Non-limiting examples of such optional ingredients include preservatives, antioxidants, emulsifiers, buffers, pharmaceutically active agents, supplemental nutrients, pigments, flavors, thickeners, stabilizers, and the like. The nutritional formula 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 formula in powder or liquid form may also contain minerals such as phosphorus, magnesium, iron, zinc, manganese, copper, sodium, potassium, molybdenum, chromium, selenium, chloride and combinations thereof. Nutritional formulas in powder or liquid form may also contain one or more flavoring agents to reduce, for example, 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 in the powdered nutritional formula may vary depending on the particular flavoring selected, the other ingredients in the formula, and other variables of the formula or the target product.

The invention will be better understood by reference to the following examples.

Examples

Example 1: conventional external fiber removal process control：

Seeds of fava beans of the same Tiffany variety are treated to separate the outer fiber and cotyledons. For this purpose, two methods are employed.

The method in the prior art comprises the following steps: seed headFirst using a knife mill (SM300,) And (6) processing. The millbase is then treated with a system known as "saw tooth" (MZM 1-40,) The treatment is carried out by turbine separation. The air flow rate is 4.0m.s^-1(23m³.h^-1). Finally, a light component containing external fibers and a heavy component containing cotyledons are obtained. The heavy components were then milled with a roller mill (MLU 202,) And (6) grinding. Finally obtaining the powder with the grain diameter less than 300 mu m. This method is illustrated in fig. 1.

According to the improved method of the invention: the seeds are first ground with a stone grinderAnd (6) processing. The millbase is then treated with a system known as "saw tooth" (MZM 1-40,) The treatment is carried out by turbine separation. The air flow rate is 4.0m.s^-1(23m³.h^-1). Finally, a light component containing external fibers and a heavy component containing cotyledons are obtained. The heavy component was then ground with a knife mill (SM300,) And (6) processing. The heavy components were then milled with a roller mill (MLU 202,) And (6) grinding. Finally obtaining the powder with the grain diameter less than 300 mu m. This method is illustrated in fig. 2.

For both processes according to the prior art and the heavy fraction obtained according to the invention described above, the residual external fibres (or "hells") therein are separated manually. This involves taking a 200g sample and then manually separating the external fibres that are still present. It is subsequently weighed (weight ═ m). The percentage of residual outer fibers is calculated by: (m/200) 100

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

Example 2: production of protein compositions according to the invention

According to [0063 ] above]The improved method according to the present invention described in the paragraph, 75kg of young broad bean powder was prepared. The powder was suspended in drinking water at 10% by weight of dry matter at 20 ℃. The pH was adjusted to 7 by adding caustic potash. Still homogenised for 15 minutes at 20 ℃. The solution was then transferred to a Sedicarter decanter from Flottweg (bowl speed: 60%, i.e. 4657rpm (about 3500g), Vr: 18.8, screw speed 60%, supernatant (overflow) was removed at 140mm by pipette at a feed rate of 1m³H), collecting the supernatant containing the protein.

The supernatant was acidified to pH 4.5 by the addition of about 7% by mass hydrochloric acid. The vessel was jacketed by injecting steam, heated to 60 ℃ and homogenized for 15 minutes. The overflow at 140mm was pipetted using again the sediscater decanter from the company Flottweg (bowl speed 60%, i.e. 4657rpm (ca. 3500g), screw speed 10% at Vr ═ 3.5 and maximum 40% (Vr ═ 12.6) until 137 had been reduced, at a feed rate of 700l/h), but this time in order to recover a precipitate containing coagulated proteins.

The precipitate was diluted to about 15% to 20% by weight of dry matter and neutralized to pH 6.5 by the addition of 20% caustic potash. The heat treatment was carried out at 135 ℃ using a nozzle, and vacuum flash cooling was carried out at 65 ℃. The product is finally atomized (inlet temperature 200 ℃ C., steam temperature 85-90 ℃ C.).

The yield of protein extracted from the meal was 72.5%. The resulting protein is referred to as a "protein composition according to the invention".

Example 3: production of protein compositions according to the prior art

This example uses the literature "structural characterization of leguminous plant protein isolates and polysaccharide gels" (textile properties of leguminous protein isolates and polysaccharide gels). (Makri et al, Journal OF THE Science OF Food AND Agriculture, 86, 1855- & 1862.) cited in THE article "influence OF GENOTYPE AND environment ON PHYSICOCHEMICAL AND FUNCTIONAL properties OF broad BEAN PROTEIN ISOLATES" (THE EFFECT OF GENOTY AND THIENVINITION ON THE PHYSOCHEMICAL AND FUNCTIONAL ATTENES OF FABA BEAN PROTEIN ISOLATES) (Shingha, 2015.). Briefly, 350g to 400g of powder were dispersed in distilled water (1: 10, w/v) and adjusted to pH 9.5 with 1M NaOH, then stirred (500rpm) at 21-23 ℃ for 40 minutes and centrifuged (1600 Xg, 20 minutes, 4 ℃). The supernatant was taken and then diluted with distilled water (1: 5, w/v), stirred and centrifuged (1600 Xg, 20 min, 4 ℃). The pH of the supernatant was adjusted to 4.5 with 1M HCl and then centrifuged (1600 Xg, 20 min, 4 ℃). The supernatant was re-diluted with deionized water, adjusted to pH 7.0 with 1M NaOH, and then lyophilized.

The yield of protein extracted from the meal was 81.2%. The resulting protein is referred to as "protein composition according to the prior art".

Example 4: functional and analytical controls

The various compositions obtained by examples 2 and 3 were compared from an analytical (dry matter and protein content) and functional (solubility according to test a) point of view. A commercially available fava bean protein composition, namely YANTAI T, 85% fava bean protein isolate from FULL BIOTECH CO LTD (lot No. DFC021606181/C1377), was also obtained, which is a representative fava bean isolate available on the market. The following table 1 summarizes these analyses.

[ Table 1]

The table shows that the protein composition according to the invention has a particularly low solubility at pH values above 7: much lower than 25% whereas the solubility of the protein composition according to the prior art exceeds 35%.

The gelling capacity of the different isolates was quantified according to the following protocol:

1. preparing an aqueous suspension by mixing water and the isolate to obtain a final suspension with a dry matter titer of 15% at a pH of 7;

2. suspension was prepared in a stress-acting rheometer equipped with a concentric column of DHR2 type (TA, instrument);

3. the elastic modulus G' and the viscous modulus G ″ were measured by applying the following temperature curves:

a. stage 1: heating from 20 deg.C to 80 deg.C within 10 min

b. And (2) stage: stabilizing at 80 deg.C for 110 min

c. And (3) stage: cooling from a temperature of 80 ℃ to 20 ℃ within 30 minutes;

the results are as follows:

it can be seen that the gelling capacity is 5 to 6 times higher than the isolates of the prior art.

Example 5: vegetarian sausage

The formulation of pea isolates sold in the formulation of the fava bean protein isolates and vegetarian food according to the control invention is as follows:

the manufacturing scheme of the sausage is as follows:

mixing water and crushed ice

Methylcellulose was dispersed into a 60% water/ice mixture using Kenwood Electronic KM231 (uk). At maximum speed for 5 minutes.

The test proteins were added and mixed using Kenwood Electronic KM231 (uk). At maximum speed for 10 minutes.

-adding the oil while stirring at maximum speed and homogenizing for another 10 minutes.

-adding the remaining powder ingredients and the remaining 40% of the water/ice mixture. Stirring was done for a final 5 minutes at maximum speed.

Pour into 2 m length of Viscofan artificial peelable cellulose casing (DATSchaub corporation, Thiais, france).

Baking in an industrial oven (Four Bourgeois S2ON 1-Ser. No. S2476057) at 100 ℃ for 1 hour with humidity control at 4.

-removing the sausage from the oven and stabilizing it at room temperature.

Manually peeling off the cellulose casing by hand.

Before analysis, the sausages were boiled in salt-free drinking water for 5 minutes and then left at room temperature for 30 minutes.

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

A test called "slicing" or "cutting" is carried out to characterize the sausage, which consists in using a texture measuring instrument to perform the action of dividing the sausage into two parts by measuring the necessary forces. This test was performed using a Warner-Bratzler cutter which penetrated the sausage completely by 25mm with a minimum detection limit of 0.06N. The maximum force at break point was used for characterization.

The values obtained were as follows:

it can be seen that the force required for slicing sausage 3 is much higher than for slicing sausage obtained from commercial pea isolate. From this result it can be concluded that the sausages obtained with the broad bean isolate according to the invention are more compact.

Example 6: tradition andlight mayonnaise

The good results of the isolates according to the invention in the preparation of traditional (called "full fat") and light (called "low fat") mayonnaises will be demonstrated below.

The raw materials required to achieve the mayonnaise formulation were as follows:

the isolate to be tested will be of the Rogai companyVicia faba isolates and aquafaba (from Vicia faba) according to the invention"Aquafaba Powder" available from the company).

The manufacturing scheme is as follows:

in HOTMIX Pro Gastro, the ingredients of stage 1 were mixed at 3 steps for 1 minute (Equipment manufacturer: MATFER-FLO, model: 212502).

Low fat formulations where the ingredients of stages 2 and 3 are added at a rate of 4 to 7 within 1 minute and 30 seconds, or full fat formulations where the ingredients of stage 2 are added at a rate of 3 within 2 minutes.

Full fat formulations, add stage 3 ingredients at 3 speeds over 1 minute.

Full fat formulations, add stage 4 ingredients at 3 speeds over 1 minute.

Complete emulsification in 1 minute, 8 speeds for low fat formulations and 3 speeds for full fat formulations.

We will compare the various mayonnaises obtained using a ta. hdplus texture tester (as shown in appendix 1) so that the firmness, consistency and cohesion parameters can be measured. Firmness (g) corresponds to the force required to be applied to force the geometric form (see "backward extrusion ring" set, described below) into the product, consistency (g.sec) is calculated from the area under the curve of firmness and cohesion (g) corresponds to the force required to be applied when the geometric form is removed from the mayonnaise.

The texture tester was equipped with a "backward extrusion ring" kit consisting of 1 disc screwed onto the equipment and 3 perspex containers for filling it with mayonnaise. The collection was done using the Exponent software, by a program designed specifically for mayonnaise analysis. The geometric mould falls to the bottom of the container at a speed of 3mm/s and rises at a speed of 5 mm/s. The software automatically plots the time-varying curves from which the parameters can be inferred.

The whole implementation is clearly explained in the user manual.

The results for the "low fat" mayonnaise were as follows:

the results for "full fat" mayonnaise were as follows:

the results obtained show that the mayonnaise obtained with the fava bean isolate according to the invention is characterized by a good texture value, which is higher in the "low fat" protein pastes than in the pea and aquafaba isolates.

Example 7: plant milk or 'milk replacer'

Vegetable milk was prepared to evaluate the performance of our isolates according to the invention in this application.

The formula is as follows:

the preparation scheme is as follows:

-heating the water to 70 ℃ and hydrating the protein isolate with Sylveson at 2000rpm for 15 minutes

Addition of other ingredients than oil and mixing for 10 minutes

-heating the oil to 65 ℃ and adding the oil with stirring at 6000rpm

UHT Sterilization at-142 ℃ for 5 seconds

Homogenizing at-75 deg.C in 2 stages (270bar and 30bar)

Cooling to 4 deg.C

The particle size distribution of the emulsified oil pellets was analyzed using a Mastersizer particle size analyzer. The particle size determination parameters were as follows: d10 ═ 0.21 micrometer, D50 ═ 0.45 micrometer, and D90 ═ 1.42 micrometer.

These results are good and well demonstrate good emulsification of the lipid globules, just like milk.

Brief description of the drawings

Other features, details and advantages of the present invention will be described in the following detailed description of the invention with reference to the drawings, in which:

FIG. 1 shows a schematic view of a

FIG. 1 shows a conventional method for separating outer fiber and cotyledon of seeds of Vicia faba;

FIG. 2

FIG. 2 shows a method for separating cotyledons from external fibers of seeds of Vicia faba according to the present invention.

Detailed Description

According to the invention, a fava bean protein composition is proposed, the colour of which is characterized by a component L higher than 70, measured according to the method of la b measurement, and a solubility lower than 25% at pH values higher than 7. Preferably, the solubility at a pH higher than or equal to 7 is lower than 25%. Even more preferably, the solubility at pH 3 is also below 25%.

According to another aspect, a method for producing a fava bean protein composition according to the invention is proposed, characterized in that it comprises the following steps: 1) providing small broad bean seeds; 2) milling seeds of fava beans using a stone mill, separating the resulting mill into two components called light and heavy using an ascending gas stream, and then secondary milling the heavy component using a knife mill; 3) final milling of the heavy fraction using a roller mill to obtain a meal; 4) suspending the powder in an aqueous solvent having a pH between 6 and 8, preferably 7; 5) removing solid components from the suspension by centrifugation and obtaining liquid components; 6) heating under isoelectric pH, and separating small broad bean protein contained in liquid component by precipitation; 7) diluting the previously obtained fava bean protein to 15-20 wt% of dry matter and neutralizing the pH to between 5.5-6.5, preferably 6.5, to obtain the fava bean protein composition; 8) drying the fava bean protein composition.

According to a final aspect, the industrial use of the fava bean protein isolate according to the invention is proposed, in particular in human or animal food, cosmetics, pharmaceuticals.

The present invention and its modifications may generally propose a practical and efficient solution to meet the need for industrially obtaining a small fava bean protein isolate, the colour of which is characterized by a component L higher than 70 and a solubility lower than 25% at pH values higher than 7, as measured according to la ab, and a method for its production and suitable industrial uses.

The invention will be better understood from the following description of the various parts.