FR3145669A1 - IMPROVED PEA OR FIELD BEAN PROTEINS - Google Patents

Abstract

The present invention relates to pea proteins having a milky aromatic profile, to a method for manufacturing these pea proteins, as well as to the use of said proteins for the manufacture of food or beverage products, in particular plant-based alternatives to milk.

Description

IMPROVED PEA OR FIELD BEAN PROTEINS Field of invention

The invention relates to a novel composition comprising pea or fava bean proteins combined with soluble fibers. Another subject of the invention relates to a method for manufacturing these novel pea protein compositions. The invention also relates to the use of said pea or fava bean protein compositions for the manufacture of food products.

Prior art

Daily protein requirements are generally between 12 and 20% of the food ration. These proteins are provided by both animal products (meat, fish, eggs, dairy products) and plant foods (cereals, legumes, algae).

In industrialized countries, protein intake is still mainly in the form of animal proteins. These proteins have good nutritional properties and interesting functional properties that allow them to be used in a wide variety of food products.

However, many studies show that excessive consumption of animal proteins to the detriment of plant proteins is one of the causes of an increase in cancers and cardiovascular diseases. Furthermore, animal proteins have many disadvantages, both in terms of their allergenicity (particularly proteins from milk or eggs), and on an environmental level linked to the harmful effects of intensive farming.

Thus, there is a growing demand from manufacturers for proteins of plant origin with interesting nutritional and functional properties, without having the disadvantages of proteins of animal origin.

Since the 1970s, the pea has been the most widely developed legume in Europe and mainly in France, particularly as a protein resource for animal and human food. The pea contains approximately 27% by weight of protein materials. The term "pea" is considered here in its broadest sense and includes in particular all wild varieties of "smooth pea", and all mutant varieties of "smooth pea" and "wrinkled pea", regardless of the uses for which said varieties are generally intended (human food, animal nutrition and/or other uses). Pea protein, mainly pea globulin, has been extracted and industrially valued for many years.

The fava bean, or fava bean (according to the old spelling), is also a well-known plant of the species Vicia faba . It is a legume of the Fabaceae family, Faboideae subfamily, Fabeae tribe. It is the same species as the broad bean, a plant used since antiquity for human consumption. The word broad bean then designates both the seed and the plant.

We can first mention the processes for extracting pea or fava bean protein called "dry". The principle of these processes is to grind the seed into flour, which will then be introduced into a turbo-separator, a device used to classify particles according to their size and density within an air flow. Turbo-separation makes it possible to obtain a protein-enriched fraction and a starch-enriched fraction. As will be developed later in the presentation, the protein-enriched fraction is approximately 40%-60% and still contains between 2% and 15% starch. This starch is a polysaccharide fraction that is not necessarily desired because it contributes to increasing blood sugar levels; its replacement by polysaccharides that are not digestible by the consumer but digestible by its digestive microflora would be of interest. These same properties are also observed in the case of fava bean.

Another example of a process for extracting pea or faba bean protein is patent EP1400537, which is characteristic of so-called “wet” extraction processes. In this process, the seed is ground in the absence of water (a process known as “dry grinding”) to obtain a flour. This flour is then suspended in water at room temperature in order to then carry out the various stages of protein extraction. This type of process separates by isoelectric precipitation the proteins belonging to the globulin subgroups (approximately 80% of pea proteins) and the proteins belonging to the albumin subgroup (approximately 20% of pea proteins). The latter remain in the liquid fraction after recovery of the floc, which is mainly composed of globulins. These albumins are in solution with the galactooligosaccharides (GOS) of the pea which are not easy for humans to digest, the salts which are mainly potassium which can be harmful in high doses as well as other antinutritional factors such as anti-trypsin factors. These same properties are also observed in the case of the field bean.

Several problems are easily guessed, such as proteins being separated into two fractions requiring the recovery of two different fractions, or GOS being combined with albumins which are difficult to digest and requiring post-treatment which complicates and makes the process more expensive. Finally, the massive presence of starch in the protein-enriched fraction of concentrates obtained by dry process, by turbo-separation, is also a nutritional disadvantage for certain formulations.

The Applicant has thus arrived, after much research, at a new manufacturing process making it possible to provide a composition which contains all of the so-called soluble proteins of peas or fava beans, i.e. comprising all of the globulins and albumins, as well as a quantity of soluble fibres derived from GOS making it possible to obtain a high-quality nutritional source.

Thus, the subject of the invention is a process for manufacturing pea or field bean protein comprising the following steps:
1. Preparation of an aqueous suspension of crushed pea or field bean seeds in an aqueous solution, said preparation being carried out, optionally, in the presence of a heat treatment;
2. Elimination of an insoluble fraction by solid/liquid separation of the aqueous suspension of crushed pea or field bean seeds obtained during step 1) allowing the production of a protein-enriched fraction;
3. Defructosylation of galactooligosaccharides of the protein-enriched fraction by enzymatic and/or fermentative means.

Preferably, the aqueous suspension of crushed pea or field bean seeds is obtained by adding a pea or field bean flour obtained by dry grinding prior to its dispersion in the aqueous solution.

When the pea or fava bean seeds are introduced in the form of whole pea or fava bean seeds into the aqueous solution, step 1 of the process comprises a step of wet grinding of the aqueous composition comprising the pea or fava bean seeds in order to obtain the aqueous suspension of ground pea or fava bean seeds.

In the absence of the optional heat treatment in step 1, the aqueous solution, as well as the resulting aqueous suspension, are at a temperature between 5°C and 30°C, preferably at room temperature. Room temperature means a temperature between 15°C and 27°C, preferably between 19°C and 23°C, the temperature values being able to be 15°C, 16°C, 17°C, 18°C, 19°C, 120°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C or 27°C.

Preferably, the optional heat treatment of step 1 comprises:
(a) introducing pea or field bean seeds or crushed pea or field bean seeds into an aqueous solution at a temperature of between 65°C and 90°C in order to obtain an aqueous composition comprising crushed pea or field bean seeds;
b) heat treatment of the aqueous composition obtained during step a) at a temperature between 40°C and 65°C for 1 to 10 min.

Preferably, the defructosylation of galactooligosaccharide (GOS) in step 3 is carried out with an enzyme selected from invertase, alpha-galactosidase, beta-fructosidase and their possible combination.

Alternatively, the defructosylation of galactooligosaccharide (GOS) is carried out with a microorganism of the genus Bacillus , preferably Bacillus subtilis , preferably a strain of Bacillus subtilis as deposited on May 28, 2020 at the CNCM under number I-5515.

Another subject of the invention relates to pea or field bean protein, characterized in that it comprises a protein content of between 50% and 70% expressed in grams of proteins per 100g of dry matter, said proteins being made up of a mixture of globulins and albumins, and between 3% and 15% of soluble fibers from pea galactooligosaccharides.

Preferably, the dry weight ratio of globulins to albumins is between 70/30 and 90/10, preferably between 75/25 and 85/15.

Preferably, the soluble fibers from pea or field bean galactooligosaccharides are selected from the list containing melibiose, manninotriose, verbascotetraose and their mixtures.

Another subject of the invention also relates to the use of said pea or field bean protein composition according to the invention for the manufacture of food or beverage products, in particular plant-based alternatives to milk.

Detailed description of the invention

The invention relates to a method for manufacturing pea or fava bean protein comprising the following steps:
1. Preparation of an aqueous suspension of crushed pea or field bean seeds in an aqueous solution, said preparation being carried out, optionally, in the presence of a heat treatment;
2. Removal of an insoluble fraction by solid-liquid separation of the aqueous suspension of crushed pea or field bean seeds from step 1) allowing a protein fraction to be obtained;
3. Defructosylation of galactooligosaccharides of the protein fraction by enzymatic and/or fermentative pathway.

Step 1) comprises the introduction of peas or fava beans into an aqueous solution. The pea or fava bean seeds used in step 1) may have previously undergone steps well known to those skilled in the art, such as in particular cleaning (removal of unwanted particles such as stones, dead insects, soil residues, etc.) or even the removal of the external fibers of the pea or fava bean (cellulosic outer shell) by a well-known step called dehulling.

Thus, by “pea” or “fava bean” in step 1), we mean whole peas or fava beans or in the form of cotyledons, from which the outer shell has been preferentially removed. Alternatively, ground pea seeds (i.e. pea flour) can be used, these ground peas generally being obtained by dry grinding.

The aqueous solution may be water which may optionally include additives such as anti-foaming compounds, salts or bacteriostatic agents.

The weight ratio of quantity of peas/quantity of aqueous solution in step 1) may in particular be between 0.1 and 2. In the embodiment where the pea or field bean seed is ground into flour before being introduced into the aqueous solution, the weight ratio of quantity of peas/quantity of aqueous solution in step 1) will be between 0.1 and 0.3, preferably between 0.15 and 0.25. Expressed as a percentage of dry matter, this ratio will be between 10% and 30%, preferably between 15% and 25%.

According to one variant, the pH of the suspension from step 1) is adjusted to between 8 and 10. This adjustment can be made by adding a base such as soda, lime or potash, preferably soda. According to another variant, the pH is not adjusted at this step.

The aqueous suspension of crushed pea or field bean seeds is obtained in step 1) by introducing into the aqueous solution, possibly previously heated to a temperature between 65°C and 90°C if heat treatment is used, the pea or field bean seeds or the crushed pea or field bean seeds.

In the case of a heat treatment, the temperature of the aqueous solution may be previously adjusted to a temperature between 65°C and 90°C. The heating may be carried out using any installation well known to those skilled in the art, such as an immersed heat exchanger. Preferably, the temperature is between 70°C and 80°C, or even approximately 75°C.

Alternatively, the aqueous solution does not undergo any prior heating and is directly at a temperature between 65°C and 90°C when mixed with the pea or fava bean seeds or the crushed pea or fava bean seeds. Preferably, the temperature of the heat treatment is between 40 and 60°C, or even between 45 and 55°C. Preferably, the heat treatment is carried out for 2 to 4 min.

In either embodiment, the method further comprises heat treating the suspension formed between the pea or fava bean seeds or the ground pea or fava bean seeds and the aqueous solution at a temperature of between 40°C and 65°C for 1 to 10 minutes. The aqueous composition comprising pea or fava bean seeds or ground pea or fava bean seeds may be heated or cooled to reach this temperature.

In the case where pea or fava bean seeds (and not already ground pea or fava bean flour) are used during the preparation of the suspension according to step 1), the method comprises a step of wet grinding of the aqueous composition comprising pea or fava bean seeds or ground pea or fava bean seeds in order to obtain an aqueous suspension of ground pea or fava bean seeds. If a heat treatment is applied, the wet grinding takes place after said heat treatment. Preferably, the method is carried out from pea or fava bean seeds and the wet grinding step is carried out by continuous passage through one or more grinders to obtain the aqueous suspension of ground pea or fava bean seeds. The grinder(s) may be any type of grinder capable of carrying out wet grinding, such as ball wet grinders, conical wet grinders, helical wet grinders or wet grinders equipped with rotor/stator systems. According to one variant, the grinder may be the one used in the examples of document WO2019/053387 in the name of the Applicant. In the variant where the grinder is of the rotor-stator type, this type of grinder may allow continuous grinding by passing the aqueous composition obtained between the pea or fava bean seeds and the aqueous solution in said grinder. According to a preferred sub-variant, the method combines two cutting steps (pre-cutting then cutting) using different rotor-stator grinders for each of these cuts. The pre-cutting then the cutting may be carried out one after the other or, alternatively, the cutting may take place after having pre-cut then stored the aqueous composition obtained between the pea or fava bean seeds and the treated aqueous solution. Such grinders are described in document WO2019/158589. Optionally, it is possible to carry out during this step or at the end of this step a dilution with water in order to form the aqueous suspension of crushed pea or field bean seeds. According to a variant, during the grinding, water is added continuously or discontinuously to dilute the aqueous composition. Generally the dry matter of the aqueous suspension of crushed pea or field bean seeds ranges from 10 to 30%, for example from 15 to 25%.

Step 2) of the method consists of extracting the components of the aqueous suspension of crushed pea or field bean seeds, and in particular extracting a protein fraction by solid-liquid separation of a so-called insoluble fraction of the aqueous suspension of crushed pea or field bean seeds. According to a variant, before carrying out the solid-liquid separation stage, it is possible to carry out a stage of adjusting the pH of the aqueous suspension of crushed pea or field bean seeds. Thus, the solid-liquid separation can take place after adjusting the aqueous suspension of crushed pea or field bean seeds to a pH ranging from 6 to 9, preferably from 8 to 9, most preferably from 8.5 to 9. This pH adjustment stage can be carried out in a stirred tank. This stage can be more or less long, and last for example from 1 to 240 minutes, generally from 5 to 60 minutes. To adjust the pH, any type of acid and/or base, organic or inorganic, or their mixtures can be added. As an example of acid, it is possible to use hydrochloric acid, sulfuric acid, citric acid or their mixtures. As an example of base, it is possible to cite soda, potash or lime and their mixtures. This addition of base or acid as well as the pH measurement can be done online. The base and/or acid can be in the form of aqueous solutions. Advantageously, before this solid-liquid separation, the aqueous suspension of crushed pea or field bean seeds is cooled to a temperature below 15°C. This temperature can in particular range from 4 to 14°C, for example from 10 to 12°C. This cooling step can be carried out by known techniques, such as passing the aqueous suspension of crushed pea or field bean seeds through a heat exchanger.

Generally, the protein fraction is the soluble part of the aqueous suspension of crushed pea or field bean seeds and the fraction rich in starch and fiber is the insoluble part. It is also possible to separate more than two insoluble fractions, and for example to recover a first insoluble fraction richer in starch and a second insoluble fraction richer in fiber. Thus, according to a variant of the process a fraction rich in starch and/or a fraction rich in fiber is recovered from the insoluble part resulting from the solid-liquid separation step 2).

By “starch-rich fraction and fiber-rich fraction” or “insoluble fraction” is generally meant a fraction comprising at least 50% starch and/or fibers. The methods for quantifying starch and fibers are known to those skilled in the art and specific methods are indicated later in the description. These fractions are conventionally recovered by known separation methods. The solid-liquid separation can in particular be carried out by means of at least one separation step with a decanter, in particular a centrifugal decanter, a centrifuge or even with hydrocyclones. The method can also make it possible to recover one or more fractions enriched in fibers and/or starch, which are eliminated from the suspension, and to recover the protein fraction useful following the method of the invention.

This step is key in differentiating with concentrates obtained by dry process. The protein-enriched fraction obtained at the end of this step contains between 0% and 0.5% starch, unlike concentrates obtained by turbo-separation which contain between 2% and 15%. This step will have a direct impact on the final composition obtained as will be discussed later in this application.

At the end of step 2), the method may comprise a step of adjusting the pH of the pea or faba bean protein to a pH ranging from 6 to 7.5, generally from 6.5 to 7.5. This step may be carried out by adding an inorganic or organic base, for example by adding sodium hydroxide. The pH is generally raised by adding a basic aqueous solution.

Step 3)

The process then comprises a step 3) of defructosylation of the galactooligosaccharides by enzymatic and/or fermentation means.

For the purposes of the present invention, the term "galactooligosaccharides" means oligomers formed from a number n of oses (monosaccharides) by alpha or beta glycosidic bond and naturally present in peas or field beans, such as raffinose or stachyose.

Preferably, the water-soluble fraction according to the invention comprises defructosylated galactooligosaccharides selected from the list containing melibiose, manninotriose and verbascotetraose.

For the purposes of the present invention, the term “melibiose” means the diholoside consisting of a galactose unit linked to a glucose unit by an α(1→6) osidic bond.

For the purposes of the present invention, the term “manninotriose” means the triholoside consisting of the chain of a galactose unit linked by an α(1→6) osidic bond to another galactose unit, itself linked to a glucose unit by another α(1→6) bond.

For the purposes of the present invention, the term “verbascotetraose”, also called “manninotetraose”, means the tetraholoside consisting of the chain of three galactose units linked by α(1→6) osidic bonds, the third galactose unit itself being linked to a glucose unit by another α(1→6) bond.

Any method well known to those skilled in the art for quantifying these defructosylated galactooligosaccharides is suitable for the purposes of the present invention. Chromatographic methods will be preferred. Preferably, those skilled in the art will use the HPAEC-PAD amperometric assay method and in particular with the following materials:
- Dionex Carbopac PA1 4*50mm pre-column - Ref. 43096
- Dionex Carbopac PA1 4*250mm column – Ref. 35391
- The detector is of the PAD type, precisely gold cell
- The eluents are:
Solvent A / NaOH 0.1M: Stir 4 liters of water under helium (flow rate: 100 ml/min) for 15 min. Add 20 ml of 50% NaOH. Stir again under helium at 40 ml/min.
Solvent B / NaOH 0.1M+0.5M sodium acetate Weigh 82g of acetate of Na directly into the can. Add 2l of H2O. Stir and under helium (flow rate: 100ml/min) for 15 min then add 10 ml of 50% NaOH, stir again and under helium. The helium flow rate can be reduced to 40 ml/min.

Standards are used to calibrate the HPLC and in particular: Reagents Reference Melibiose Fluka ref 63630 Raffinose Sigma ref R-0514 Stachyose Sima ref S-4001 Verbascose Fluka ref 56217

We also use an internal standard: Panose ref SIGMA P-2407 60mg in 100ml of water.

The injected volume is 5µl at a temperature of 15°C. The analysis time is 90 min with a column temperature of 30°C and an injector sensitivity of 300nC or 5µA

The chromatographic elution conditions are as follows: Time (min) Flow rate (ml/min) Solvent A Solvent B 0 0.5 98 2 60 0.5 95 5 65 0.5 70 30 65.05 0.5 0 100 75 0.5 0 100 75.05 0.5 98 2 90 0.5 98 2

The oxidation program of the PAD detector is as follows: Time (min) Potential Integration 0 +0.05 0.20 +0.05 Beginning 0.40 +0.05 END 0.41 +0.75 0.60 +0.75 0.61 -0.15 1.0 -0.15

Calibration is performed by preparing curves according to the table below: Quantity in mg Melibiose
QSP 50ml Raffinose
QSP 25ml Stachyose
QSP 25ml Verbascose
QSP 25ml T1 10 5 5 5 T2 25 10 10 10 T3 50 15 15 15 T4 75 25 25 25 T5 100

Take 1 ml of control (from the 2 curves) + 1 ml of internal standard, qsp 20 ml of water.

Weigh the amount in mg of sample, add 1 ml of internal standard and adjust to 20 ml of water.

Filter on GxF/GHP 0.45 µm ref 4559T.

In a first alternative, the defructosylation of galactooligosaccharide (GOS) is carried out with an enzyme selected between invertase, alpha-galactosidae, beta-fructosidase and their possible combination.

Preferably, the enzyme used is an invertase such as Sumizyme INV. The pH and temperature are adjusted to optimize the defructosylation reaction, such as pH 5 and 55°C for Sumizyme INV. The reaction is controlled by performing an analysis, for example by HPLC, of the sugars present in solution. When the defructosylation is sufficient or even complete, it is neutralized using bases such as sodium hydroxide and then a heat treatment is performed to inhibit the enzyme, for example 130° for 10 seconds.

In a second alternative, the defructosylation of galactooligosaccharide (GOS) is carried out with a microorganism of the genus Bacillus , preferentially Bacillus subtilis , preferentially a strain of Bacillus subtilis as deposited on May 28, 2020 at the CNCM under number I-5515.

According to the invention, the term "fermentation" means metabolic processes that generally convert carbohydrates into acids, gases or alcohols to extract part of the chemical energy while reoxidizing the coenzymes reduced by these reactions. This is a redox metabolic pathway in which the ultimate electron acceptor is often confused with the end product of the reactions. It is characterized by a partial degradation of the fermentable substance and allows only limited energy production. It takes place in yeasts and bacteria, as well as in muscle cells lacking oxygen, i.e. under anaerobic conditions.

It is important in this process step to defructosylate the GOS without altering their carbon skeleton, not to hydrolyze it. Some prior art solutions to solve this technical problem are used. The present invention prefers to transform them into fibers beneficial for human and animal nutrition.

Steps 1), 2) and 3) of the method are carried out in this order. However, other optional steps may be implemented between steps 1), 2) and 3), such as a pH adjustment, if this is necessary in the eyes of the person skilled in the art. Preferably, after the defructosylation step, the method may comprise a step 4) of additional heat treatment of the pea or faba bean protein. The temperature and time conditions may vary widely in this step, for example ranging from 70 to 140°C and lasting from 0.1 seconds to several minutes. According to a first variant of this additional heat treatment step, the temperature ranges from 70 to 90°C and its duration ranges from 0.1 seconds to 30 minutes. According to a second variant of this additional heat treatment step, the temperature ranges from 90 to 110°C and its duration ranges from 0.1 seconds to 5 minutes. According to another variant, this additional heat treatment step is carried out at a temperature ranging from 110 to 140°C for a time ranging from 0.1 to 30 seconds, preferably from 0.2 to 15 seconds, for example from 0.3 to 10 seconds. This step may aim to functionalize and/or sanitize the pea protein. To carry out this additional heat treatment step, the pea or faba bean protein may be in the form of an aqueous dispersion, preferably having a dry matter ranging from 10 to 25%, for example from 15 to 20%. Advantageously, the method of the invention comprises, following the additional heat treatment step, a step of cooling the pea protein. According to a preferred variant, this cooling step is obtained by rapid cooling (“flash cooling”). At the end of this step, the temperature may range from 60 to 100°C, for example between 70 and 90°C. Similarly, this flash-cooling step is achieved by applying a vacuum to the aqueous pea protein dispersion, the vacuum applied being determined according to the chosen cooling temperature.

According to a variant of the method, it comprises an optional step 5) of shearing the pea or faba bean protein, for example by passing the aqueous dispersion of proteins through a high-pressure pump. As an example of a high-pressure pump, it is possible to cite the high-pressure pumps marketed by the company Silverson, also called high-shear mixers, for example those in the UHS range. Preferably, the shearing step is carried out by a high-pressure pump.

The shearing step can take place before or after the heat treatment and/or pH raising steps.

According to another variant, the method alternatively comprises an optional step 5) of homogenization of the pea protein.

To carry out this homogenization step, it is possible to use any type of homogenizer. According to the invention, it is understood to mean equipment comprising a high-pressure pump and a homogenization head in which the equipment is designed so that the product to be homogenized passes under pressure through this homogenization head. A homogenization head consists of a reduced orifice generally comprising a seat, a valve and a shock ring. The passage of the aqueous dispersion of pea or faba bean proteins through the homogenizer can thus allow the homogenization of the pea protein. The homogenization can be low-pressure homogenization, high-pressure homogenization or even ultra-high-pressure homogenization. The homogenization pressure can vary widely and range, depending on the homogenization technique used, from 1 to 1000 bar, for example from 20 to 800 bar. According to one variant, the homogenization pressure ranges from 20 to 200 bar, for example from 50 to 150 bar. According to another variant, the homogenization pressure ranges from 200 to 800 bar, for example from 300 to 800 bar. According to one variant, the homogenization is a single-effect homogenization. According to another variant, the homogenization is a multiple-effect homogenization, for example a double-effect homogenization. The homogenizers that can be used are marketed, for example, by the company GEA or Tetra Pak.

The homogenization step can take place before or after the heat treatment and/or pH raising steps.

The method according to the invention may also comprise an optional step 6) of membrane nanofiltration preferably carried out with a cut-off threshold of between 150 Da and 300 Da. This step makes it possible to pre-concentrate the composition before the potential subsequent steps of evaporation and/or drying but also to desalinate. Tests have shown that with a cut-off threshold of 300 Da the ash content can be reduced from 8 to 10% initially to values of 5 to 7%.

In order to carry out the measurement of the ash content, the person skilled in the art will use any method well known in the field. Preferably, the person skilled in the art will proceed as follows:
- Weigh a sample P1 pea
- Place the sample in an oven at 550°C for 24 hours.
Weigh the new sample weight P2
Ash content = (P2/P1)*100.

The method according to the invention may also comprise an optional step 7) of drying the pea or fava bean protein. Generally, this drying step is carried out so as to achieve a dry matter content greater than 80%, preferably greater than 90%, most preferably greater than 94% by weight of dry matter relative to the weight of the pea protein. Any technique well known to those skilled in the art is used for this purpose, such as freeze-drying, flash drying or drying on a drying drum or even atomization. The method may also comprise a grinding or micronization step. Atomization is the preferred technology, in particular multiple-effect atomization. The pea or fava bean protein may be in the form of a powder having a particle size d50, which may vary widely, for example from 10 to 500 µm, generally from 50 to 150 µm.

By “d50” is meant in the present invention the particle size measured in micrometers separating into two populations in number containing respectively 50% and 50% of the total particles of the protein composition.

To perform this measurement of d50, a laser granulometer is preferably used, even more preferably the Mastersizer 2000 from Malvern. The parameters used are as follows: Use in liquid form, dispersion in ethyl alcohol; Refractive index: 1.52; Absorption index: 0.1; no use of ultrasound.

Preferably, drying step 7) may comprise a first sub-step of concentration by evaporation followed by the actual drying step. This evaporation is conventionally carried out using any appropriate technique well known to those skilled in the art, such as vacuum evaporation. The target dry matter content is between 15% and 30%, preferably between 20% and 23%. This evaporation will make it possible to both eliminate undesirable volatile compounds and optimize the drying step.

Steps 1) to 7) of the process can be carried out in this precise order but, depending on the needs of the person skilled in the art, other optional steps can be implemented, such as a pH adjustment.

Pea or fava bean protein

Another subject of the invention relates to a pea or field bean protein composition capable of being obtained by the process of the invention.

Another subject of the invention relates to the pea or field bean protein composition characterized in that it comprises a protein content of between 50% and 70% expressed in grams of proteins per 100g of dry matter, said proteins being made up of a mixture of globulins and albumins, and between 3% and 15% of soluble fibers from pea galactooligosaccharides.

By “pea protein composition” is meant a composition that comprises primarily, but not exclusively, pea protein. Such a composition may have residual impurities such as, for example, minerals, sugars, etc.

By “fava bean protein composition” is meant a composition that comprises primarily, but not exclusively, fava bean proteins. Such a composition may have residual impurities such as, for example, minerals, sugars, etc.

The term "pea" is to be understood in the present application as all wild varieties of "smooth pea", and all mutant varieties of "smooth pea" and "wrinkled pea". The term "pea" is here considered in its broadest sense and includes in particular all varieties of "smooth pea" and "wrinkled pea", and all mutant varieties of "smooth pea" and "wrinkled pea", regardless of the uses for which said varieties are generally intended (human food, animal nutrition and/or other uses). The term "pea" in the present application includes varieties of pea belonging to the genus Pisum and more particularly to the species sativum and aestivum . Said mutant varieties include in particular those called "r mutants", "rb mutants", "rug 3 mutants", "rug 4 mutants", "rug 5 mutants" and "lam mutants" as described in the article by CL HEYDLEY et al. entitled "Developing novel pea starches" Proceedings of the Symposium of the Industrial Biochemistry and Biotechnology Group of the Biochemical Society, 1996, pp. 77-87.

The term "protein" should be understood in the present application as macromolecules formed from one or more polypeptide chains consisting of the chain of amino acid residues linked together by peptide bonds. In the particular context of pea proteins, the present invention relates more particularly to globulins (approximately 50-60% by weight of pea proteins) and albumins (20-25% by weight of pea proteins).

For the purposes of the present invention, the term "globulins" means proteins soluble in neutral saline solutions. Pea or field bean globulins are mainly subdivided into three subfamilies: legumes, vicilins and convicilins.

For the purposes of the present invention, the term “albumin” means proteins soluble in pure water. Pea albumins, present in pea or field bean proteins at a level of approximately 20%, are mainly subdivided into two families called PA1 and PA2. Pea or field bean albumins are mainly subdivided into two families called PA1 and PA2.

Generally, the protein content by weight of the pea or field bean protein according to the invention is between 50% and 70% expressed in grams of protein per 100g of dry matter. The protein content is the N6.25 content, calculated by the Dumas method. Preferably, the protein content is between 50% and 70%, comprising 50%, 51%, 52%, 53% 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62% 63%, 64%, 65%, 66%, 67%, 68%, 69 or 70%.

The pea or field bean protein according to the invention obviously generally comprises other minority constituents other than proteins, such as starch, lipids, fibers, and/or sugars.

Generally, the total starch content in the pea or field bean protein produced according to the method of the invention is between 0% and 0.5%, for example 0.1% to 0.4%, in particular 0.2 to 0.3%. This total starch content can be measured using the AOAC 996.11 method. The residual starch contents may therefore be 0%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%. This particularly low starch content is characteristic of the pea or field bean protein according to the invention. Pea or fava bean protein concentrates obtained by turboseparation contain more starch, for example between 2 and 5% according to the Feedipedia web page (https://www.feedipedia.org/node/7439) but more often up to more than 10% as will be exemplified later in this application in commercial concentrates. The presence of this residual starch will impact the nutritional character of the product but also the functional character such as for example a greater viscosity when heated.

Typically, the total lipid content ranges from 0 to 15%, for example from 1 to 10%. The total lipid content can be determined by AOAC method 996.06 in acid hydrolysis.

Preferably, the dry weight ratio of globulins to albumins is between 70/30 and 90/10, preferably between 75/25 and 85/15.

Preferably, the pea or field bean protein according to the invention has a degree of hydrolysis, or DH, of between 6% and 8%, preferably between 6.5% and 7.5%. The values of degree of hydrolysis will thus be 6.0%; 6.1%; 6.2%; 6.3%; 6.4%; 6.5%; 6.6%; 6.7%; 6.8%; 6.9%; 7.0%; 7.1%; 7.2%; 7.3%; 7.4%; 7.5%; 7.6%; 7.7%; 7.8%; 7.9% or 8.0%.

By “degree of hydrolysis” is meant in the present invention the percentage ratio between the quantity of amine (or carboxylic) functions of the free amino acids on the total quantity, including the free functions and those engaged in a peptide bond (chemical bond characteristic of proteins resulting from the association of a carboxylic function of a first amino acid and an amine function of a second). For a protein composition constituted by the chaining of all its amino acids and therefore having only one free amine function and one free carboxylic function, this degree of hydrolysis will be 0%. Conversely, for a protein composition of which the same amino acids are all said to be “free” i.e. of which their two amine and carboxylic functions are not involved in peptide bonds, this degree of hydrolysis will be 100%.

There are several methods for quantifying the degree of hydrolysis. They all consist mainly of the colorimetric assay of the free amine (or carboxylic) functions, then the performance of a hydrolysis aimed at destroying all of the peptide bonds and finally a colorimetric assay of the total amine (or carboxylic) functions. The percentage calculated between the free amines (or carboxylic) in relation to the total gives the degree of hydrolysis. Any well-known method may be used, such as the so-called TNBS method or the OPA method. In the present invention, the OPA method is preferred, a measurement procedure for which is described below:

First, the amino nitrogen content (free _NH2 ) is determined on the protein sample according to the invention using the MEGAZYME kit (reference K-PANOPA). The protein nitrogen content (total nitrogen) of the sample is also determined. It is then possible to calculate the degree of hydrolysis.

Determination of amino nitrogen content:

The “amino nitrogen” groups of the free amino acids in the sample react with N-acetyl-L-cysteine and OPhthaldialdehyde (OPA) to form isoindole derivatives.

The amount of isoindole derivative formed during this reaction is stoichiometric with the amount of free amino nitrogen. It is the isoindole derivative that is measured by the increase in absorbance at 340 nm.

In a 100 mL beaker, a test portion P* is introduced, weighed exactly, of the sample to be analyzed. This test portion will be 0.5 to 5.0 g depending on the amino nitrogen content of the sample. Approximately 50 mL of distilled water is added, homogenized and transferred to a 100 mL graduated flask. 5 mL of 20% sodium dodecyl sulfate (SDS) is added and the volume is completed with distilled water to reach a volume of 100 mL. Stir for 15 minutes with a magnetic stirrer at 1000 rpm. Solution No. 1 is prepared by dissolving a tablet from bottle 1 of the Megazyme kit in 3 mL of distilled water and stirring until completely dissolved. One tablet is required per test. Solution No. 1 is prepared extemporaneously.

A blank, a standard and a sample are prepared directly in the spectrophotometer cells under the following conditions:
- blank: introduce 3.00 ml of solution no. 1 and 50 μl of distilled water
- standard: introduce 3.00 ml of solution no. 1 and 50 μl of bottle 3 of the Megazyme kit
- sample: introduce 3.00 ml of solution no. 1 and 50 μl of the sample preparation.

The contents of each tank are mixed and the absorbance measurement (A1) of the solutions is read after approximately 2 minutes using a spectrophotometer at 340 nm (spectrophotometer equipped with tanks with a 1.0 cm optical path, capable of measuring at a wavelength of 340 nm, and checked according to the operating procedure described in the manufacturer's technical manual relating to it).

The reactions are then initiated immediately by adding 100 μl of solution no. 2 which corresponds to the OPA solution from bottle 2 of the Megazyme kit to each spectrophotometer cuvette.

The contents of each tank are mixed and placed in the dark for about 20 minutes.

The absorbance measurement A2 of the blank, the standard and the sample is then read on the spectrophotometer at 340 nm.

The free amino nitrogen content, expressed as a percentage by weight relative to the weight of the product, is given by the following formula:
Or :
ΔAech =Aech2 – Aech1
ΔAblc =Ablc2 – Ablc1
Aech2 = absorbance of the sample after addition of solution no. 2
Aech1 = absorbance of the sample after addition of solution no. 1
Ablc2 = absorbance of the blank after addition of solution no. 2
Ablc1 = absorbance of the blank after addition of solution no. 1
V = volume of the vial
m = mass of the test sample in g
6803 = extinction coefficient of the isoindole derivative at 340 nm (in L.mol ^-1 .cm ^-1 ).
14.01 = molar mass of nitrogen (in g.mol ^-1 )
3.15 = final volume in the tank (in mL)
0.05 = test portion in the tank (in mL)

Determination of protein nitrogen content:

The protein nitrogen content is determined according to the DUMAS method according to ISO 16634 - 2016. It is expressed as a percentage by weight relative to the weight of the product.

Calculation of the degree of hydrolysis

The degree of hydrolysis (DH) is calculated with the following formula:

The pea or field bean protein according to the invention contains between 3% and 15% of soluble fibers, preferably between 4% and 8%, from pea galactooligosaccharides. The content can therefore be between 3%, 4%, 5%, 6% 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% and 15%.

Preferably, the soluble fibers from pea or field bean galactooligosaccharides are selected from the list containing melibiose, manninotriose, verbascotetraose and their mixtures.

The combination of the protein, composed of a mixture of globulin and albumin, and the soluble defructosylated fibers allows the obtaining of a unique ingredient of certain interest. With this single ingredient, it is possible to provide both a quality protein and a quantity of soluble fibers. Furthermore, according to the thesis “PREVENTION AND TREATMENT OF IRON DEFICIENCY ANEMIA IN WOMEN AND CHILDREN: IRON HOMEOSTASIS AND OPTIMIZING ABSORPTION USING PREBIOTICS AND BREAST MILK COMPONENTS” (Giorgetti, 2022), iron absorption is increased when consumed with GOS.

The properties of pea or fava bean protein can vary widely, depending on the process parameters outlined above and as shown in the Examples section.

According to one embodiment, the dried pea or fava bean protein has a solubility at pH 7 ranging from 10 to 99%. The solubility can have all intermediate amounts (i.e. 11%, 12%, 13%... 97%, 98%, 99%) and the person skilled in the art will know, on the basis of the process indications indicated above and in the Examples section, how to modify the process parameters in the ranges indicated in order to achieve the desired solubility. Advantageously, the dried pea or fava bean protein has a solubility at pH 7.0 and at pH 4.0 ranging from 40% to 55%.

Solubility: Test A

As for solubility, it is determined according to the TEST A method described below:
Measuring water solubility
This measurement is based on diluting the sample in distilled water, centrifuging it and analyzing the supernatant.

Operating mode:
In a 400 ml beaker, introduce 150 g of distilled water at a temperature of 20°C +/- 2°C, mix with a magnetic bar and add precisely 5 g of the sample to be tested.
Adjust or not the pH to the desired value with NaOH or 0.1 N HCl (pH 7).
Add water to 200 g.
Mix for 30 minutes at 1000 rpm and centrifuge for 15 minutes at 3000 g.
Collect 25 g of the supernatant.
Place in a previously dried and tared crystallizer.
Place in an oven at 103°C +/- 2°C for 1 hour.
Then place in a desiccator (with desiccant) to cool to room temperature and weigh.

The soluble solids content, expressed in % by weight, is given by the following formula:

[Math. 1]

Or :
P = weight, in g, of the sample = 5 g
m1 = weight, in g, of the crystallizer after drying
m2 = weight, in g, of the empty crystallizer
P1 = weight, in g, of the collected sample = 25 g

According to one embodiment, the dried pea or field bean protein may have a gelling power. This gelling power may range from 1 to 100 Pa, preferably between 10 and 90 Pa, even more preferably between 30 and 60 Pa.

Gelling power: Test B

By "gelling power" is meant the functional property consisting of the ability of a protein composition to form a gel or a network, increasing the viscosity and generating a state of matter intermediate between the liquid and solid states. It is also possible to use the term "gel strength". To quantify this gelling power, it is therefore necessary to generate this network and to evaluate its strength. To carry out this quantification, in the present invention, test B is used, the description of which is as follows:
1) Solubilization at 60°C +/- 2°C of the tested protein composition in water titrating 15% +/- 2% in dry matter and at pH 7;
2) Stir for 5 min at 60°C +/- 2°C;
3) Cooling to 20°C +/- 2°C and stirring for 24 hours at 350 rpm;
4) Implementation of the suspension in an imposed stress rheometer equipped with a concentric cylinder;
5) Measurement of elastic moduli G' and viscous moduli G'' by applying the following temperature profile:
a. Phase 1: Measurement of parameter G'1 after stabilization at 20°C +/- 2°C and heating from a temperature of 20°C +/- 2°C to a temperature of 80°C +/- 2°C in 10 minutes;
b. Phase 2: stabilization at a temperature of 80°C +/- 2°C for 110 minutes; c. Phase 3: cooling from a temperature of 80°C +/- 2°C to a temperature of 20°C +/- 2°C in 30 min and measurement of G'2 after stabilization at 20°C +/- 2°C;
6) Calculation of the gelling power equal to G'2 – G'1.

Preferably, the imposed stress rheometers are chosen from the DHR 2 (TA, instruments) and MCR 301 (Anton Paar) models, with a concentric cylinder type mobile. They have a Peltier effect temperature control system. In order to avoid evaporation problems at high temperatures, paraffin oil is added to the samples.

A “rheometer” within the meaning of the invention is a laboratory device capable of making measurements relating to the rheology of a fluid or gel. It applies a force to the sample. Generally of small characteristic dimension (very low mechanical inertia of the rotor), it allows the fundamental study of the mechanical properties of a liquid, gel, suspension, paste, etc., in response to an applied force.

The so-called "imposed constraint" models allow, by applying a sinusoidal stress (oscillation mode), to determine the intrinsic viscoelastic quantities of the material, which depend in particular on time (or the angular speed ω) and the temperature. In particular, this type of rheometer allows access to the complex module G*, itself allowing access to the modules G' or elastic part and G'' or viscous part;

The first three steps consist of resuspending the protein in water, under precise conditions to maximize the subsequent measurement.

The water chosen is preferably osmosis water, but it is also possible to use drinking water.

Its temperature is 60°C +/- 2°C during the initial resuspension (1st and 2nd steps) then 20°C +/- 2°C after solubilization for 24 hours and cooling before measurement (3rd step). Generally speaking and unless otherwise indicated, when a temperature is given in this description, it always includes a variation of +/- 2°C, for example 20°C +/- 2°C or 80°C +/- 2°C.

A defined quantity of protein is added to said water in order to obtain a suspension titrating 15% +/- 2% in dry matter. To do this, equipment well known to those skilled in the art is used, such as beakers and magnetic bars. A volume of 50 mL is stirred for at least 10 hours at 350 rpm at room temperature. Generally speaking and unless otherwise indicated, the dry matter contents given in the present description always include a variation of +/- 2%, for example 15% +/- 2%. The pH is adjusted to 7 +/- 0.5 using a pH meter and acid-base reagents, as well known in the prior art.

The fourth step is to introduce the sample into the rheometer, covering it with a thin layer of oil to limit evaporation.

The following temperature scale is then applied in the fifth step: a. Phase 1: heating from a temperature of 20°C +/- 2°C to a temperature of 80°C +/- 2°C in 10 minutes; b. Phase 2: stabilization at a temperature of 80°C +/- 2°C for 110 minutes; c. Phase 3: cooling from a temperature of 80°C +/- 2°C to a temperature of 20°C +/- 2°C in 30 min.

The measurement of the G’ parameter is carried out continuously during this scale and is recorded.

The sixth and final step of test B consists of the exploitation of the recording. Two values are extracted: G’1 = value of G’ at the beginning of phase 1 after stabilization at 20°C +/- 2°C and G’2 = value of G’ at the end of phase 3 after stabilization at 20°C +/- 2°C.

The gelling power is equal to G’2 – G’1.

According to an optional variant, the pea or faba bean protein is an enzymatically modified protein. By enzymatically modified protein, the person skilled in the art means a protein whose protein structure has been deliberately modified by adding to the protein at least one enzyme capable of modifying the protein structure. This enzyme can be chosen from proteases, peptidases, deamidation enzymes, for example those of type E.C. 3.5.1 such as glutaminase or deimination enzymes, for example those of type E.C. 3.5.3 such as peptidylarginine deiminase. These protein modification enzymes are known to modify the physicochemical and/or organoleptic properties of the protein. For example, it is known from document WO2019/233920 A1 that peptidylarginine deiminase makes it possible to reduce astringency, in particular the astringency of rapeseed protein. In the case where the method comprises proteolysis of the protein fraction enriched in proteins, this can make it possible to modify the degree of hydrolysis (DH) of the protein. Preferably, the degree of hydrolysis is less than 15%, advantageously less than 10%, preferably less than 6%, for example between 3 and 5%. A person skilled in the art will know how to adapt the conditions of enzymatic proteolysis, or will even not carry out such a step in order to obtain the desired DH. According to a preferred variant of the invention, the pea or fava bean protein is not enzymatically modified by deamination. According to another preferred variant of the invention, the protein is not enzymatically modified. An advantage of the invention is that it is possible to modify the organoleptic properties of the pea protein, and in particular to give it a milky aromatic universe, without even needing to enzymatically modify the protein. The invention therefore makes it possible to provide, according to one embodiment, proteins with an unmodified primary structure.

Uses of pea protein

The invention also relates to the use of the pea or field bean protein of the invention for the manufacture of food or beverage products, in particular plant-based alternatives to milk.

Generally, the pea or faba bean protein of the invention may be used in food and beverage products which may include it in an amount of up to 100% by weight relative to the total dry weight of the food or beverage product, for example in an amount of from about 1% by weight to about 80% by weight relative to the total dry weight of the food or beverage product. Any amount in between (i.e. 2%, 3%, 4%... 77%, 78%, 79% by weight relative to the total weight of the food or beverage product) may be used, as may any intermediate ranges based on these amounts. Such food and beverage products may be suitable for vegetarian or vegan populations.

A particularly interesting use of the protein of the invention concerns its use in beverages which have a more pleasant taste than those obtained from other commercial pea proteins. The pea or fava bean protein of the invention can advantageously be used for the manufacture of beverages, in particular alternatives to milk, or in other words milk substitutes. Furthermore, due to the milky aromatic note due to the ingredient, these beverages can also have a more milky aromatic note than a beverage not comprising said protein, which is an undeniable advantage for the manufacture of plant-based alternatives to milk. In addition to an improvement in the aroma, it is also possible according to the invention to obtain a more coating texture in the mouth (mouthfeel effect) than when other pea or fava bean proteins are used, which is an advantage for beverages, and in particular for plant-based alternatives to milk because animal milks generally also have a coating texture.

In beverages, the protein content in these products can vary widely and can also be a high protein drink. The amount of protein can range for example from 1 to 12% in dry mass relative to the total mass of the beverage, in particular from 3 to 10% relative to the total mass of the beverage. The beverages can be of any type and include plant-based alternatives to milk or milk substitutes, including "barista" type milks or "coffee creamers". It may also include other ready-to-drink beverages, whether acidic or not, such as carbonated beverages (including, but not limited to, carbonated soft drinks), non-carbonated beverages (including, but not limited to, non-carbonated soft drinks such as flavored waters, fruit juices, and sweetened or unsweetened tea or coffee-based beverages), alcoholic beverages such as beers or spirits, smoothies, beverage concentrates (including, but not limited to, liquid concentrates and syrups, and non-liquid “concentrates,” such as freeze-dried and/or powdered preparations or “powder mixes”). Note that in beverages, flavorings or masking agents are generally used to reduce the pea or fava bean flavor note or the bitter aftertaste of the protein or to flavor the beverage. One of the advantages of the pea or fava bean protein of the invention is that its use in place of conventional pea or fava bean proteins makes it possible to reduce this quantity of flavoring or masking agent, or even to completely eliminate these constituents from the beverage, while maintaining a very satisfactory taste for the beverage. The beverages may also include hydrocolloids; however, since the pea or fava bean protein provides a more coating texture, it is possible to reduce or even eliminate the content of hydrocolloid agents while maintaining a coating texture in the mouth.

Food products that may be affected include bakery products such as bread products (including, but not limited to, sourdough and unleavened breads, sandwich breads, yeast breads and yeast-free breads such as soda breads), breads made from all types of wheat flour, breads made from all types of flour other than wheat flour (such as potato, rice, barley, spelt and rye flours), gluten-free breads; mixes for the preparation of such bread products; sweet bakery products (including, but not limited to, rolls, cakes, pies, pastries, waffles, crepes, muffins, pancakes, and biscuits); mixes for the preparation of such sweet bakery products; Pie fillings and other sweet fillings (including, but not limited to, fruit pie fillings and nut pie fillings such as pecan pie fillings, as well as fillings for cookies, cakes, pastries, confectionery products and the like, such as cream fillings); snack bars (including, but not limited to, energy, granola, nut, and/or fruit bars).

It can also be gelled desserts such as puddings or custards and puddings. Another type of dessert can also be frozen desserts (including, but not limited to, frozen dairy desserts such as ice cream - including regular ice cream, soft serve ice cream and all other types of ice cream - and frozen non-dairy desserts such as non-dairy ice cream, sorbet and others).

Other products conventionally prepared from animal milk may also include the pea or faba bean protein of the invention to form substitutes. These may be acidified products and/or fermented with ferments, for example lactic, vegan or mesophilic ferments. These may be yogurts (including, but not limited to, full-fat, reduced-fat and fat-free yogurts, which yogurts may be free of milk proteins and lactose-free). The term “yogurts” also includes fromage blanc and petits suisses. They may also be cheese substitutes such as spreadable, processed, cooked and uncooked pressed cheeses, soft cheeses, stretched cheeses, blue cheeses; These include Emmental, String Cheese, Ricotta, Provolone, Parmesan, Munster, Mozzarella, Monterey Jack, Manchego, Blue, Fontina, Feta, Edam, Double Gloucester, Camembert, Cheddar, Brie, Asiago and Havarti. They can also include other products such as vegetable butters or crème fraîche.

Other products that may include the pea or fava bean protein of the invention are also sauces such as vinaigrettes or mayonnaise or ketchup-based sauces or syrups.

Also, the pea or faba bean proteins of the invention can be incorporated into confectionery products (including, but not limited to, gummy candies, soft candies, hard candies, chocolates, caramels and gums); sweetened and unsweetened breakfast cereals (including, but not limited to, extruded cereals, flaked cereals and puffed cereals); and cereal coating compositions for preparing breakfast cereals. They can also be sweetened spreads (including, but not limited to, jellies, jams, nut butters such as peanut butter, spreads and other spreadable products).

The pea or field bean proteins of the invention can also be used as a carrier or in flavor encapsulation.

Other types of foods and beverages not mentioned herein but which typically comprise one or more proteins and may also be contemplated within the scope of the present invention. In particular, animal foods (such as pet foods) are explicitly contemplated.

Pea or fava bean protein can also be used, possibly after texturing, in meat substitutes such as emulsified sausages or hamburgers, or fish or seafood substitutes. It can also be used in egg replacement formulations or for the manufacture of protein products such as tofu or tempeh. Textured proteins generally mean proteins textured by extrusion, i.e. in particular dry extrusion (“dry extrusion” or “Textured Vegetable Protein”), wet extrusion (“high moisture extrusion”). The extruders can be single-screw, twin-screw or multiple-screw extruders. In the case of twin-screw extrusion, the extrusion can be co-rotating or counter-rotating. Examples of multiple-screw extrusion include the planetary extruder or the ring extruder. It is also possible to cite other more specific technologies such as “shear cell” technology, microextrusion or even 3D printing.

Food or beverage products may be used in particular in specialized nutrition, for example for specific populations, for example for babies or infants, children, adolescents, adults, the elderly, athletes, people suffering from a disease. They may be meal replacement nutritional formulas, complete nutritional drinks, for example for weight management or in clinical nutrition (for example tube feeding or enteral nutrition).

Pea or faba bean protein can be used as the sole source of protein, but can also be used in combination with other additional proteins, plant or animal. These additional proteins can be hydrolyzed or non-hydrolyzed. Generally, these additional proteins are in the form of concentrates or isolates. Concentrates are distinguished from isolates according to their protein content: concentrates whose protein contents are generally between 50% and 70% and isolates whose protein contents are greater than 70%, preferably between 80% and 90%, respectively. The term "plant protein" designates all proteins derived from cereals, oilseed plants, legumes and tuberous plants, as well as all proteins derived from algae and microalgae or fungi, used alone or in a mixture, chosen from the same family or from different families. By "legume" is generally meant the family of dicotyledonous plants of the order Fabales. Several legumes are important cultivated plants among which soybean, beans including mung bean, chickpea, fava bean, peanut, cultivated lentil, cultivated alfalfa, various clovers, broad beans, carob, licorice and lupin. The additional legume protein can be selected from these legumes or be a pea or fava bean protein than that of the invention. In the present application, the term "cereals" designates cultivated plants of the family of grasses producing edible grains, for example wheat, oats, rye, barley, corn, sorghum or rice. The tubers can be carrot, cassava, konjac, potato, Jerusalem artichoke, sweet potato. Oilseed plants are generally plants producing seeds from which oil is extracted. The oil plants may be chosen from sunflower, rapeseed, peanut, sesame, squash or flax. Animal proteins may be, for example, egg or milk proteins, such as whey proteins, casein or caseinates. The pea or field bean protein composition of the invention may thus be used in combination with one or more of these proteins or amino acids in order to improve the nutritional properties of the final product, for example to improve the PDCAAS of the protein or to provide other functionalities.

Pea or fava bean protein can also be used for the manufacture of pharmaceutical products or in fermentation, for example for the production of fungal metabolites or metabolites by cell culture.

The invention and its advantages will now be illustrated in the embodiments detailed in the examples section below. It is specified that these examples are not limiting of the present invention.

Examples

Example 1: P concentrate pea protein

Approximately 1000 kg of peas were used. The outer fibres of the peas were first separated from the seeds by crushing (mechanical separation of the outer shell and the pea seed) and dehulling (sorting of the outer shells and dehulled pea seeds using compressed air). The seeds thus prepared were ground using an attrition mill to obtain a particle size such that 88% of the particles were less than 100 microns and 1.9% were greater than 315 microns. The flour thus obtained was introduced into water so as to obtain a suspension of ground pea seed flour of approximately 20% dry matter. This suspension of ground pea seeds was stirred for 5 minutes and fed into a centrifugal decanter (Flottweg Z3). The protein-enriched fraction was recovered in the overflow (approximately 7% dry matter).

The protein-enriched fraction was adjusted to pH 6.6 using sodium hydroxide and hydrochloric acid, then heat-treated at 130°C for 10 seconds and flash-cooled to approximately 60°C. The solution was then atomized on a Nubilosa atomizer (air inlet temperature = 195°C – air outlet temperature = 95°C). The pea protein powder named “Pea concentrate according to example 1” was then analyzed.

Pea concentrate
according to example 1 NUTRALYS® F85M
(Pea Protein Isolate) VESTKORN Pea Protein Concentrate Pea protein concentrate
marketed by LA-VITA under the name “pea protein concentrate” Proteins 62.0% 85.10% 54.0% 50-55% Carbohydrates 24.0% 4.2% 30.6% 12-15% including starch 0.1% 0.1% 5.4% 12-15% including glucose 1.6% < 0.1% < 0.1% < 0.1% including fructose 0.2% < 0.1% < 0.1% < 0.1% including sucrose 7.8% < 0.1% 2.7% < 0.1% whose melibiose < 0.1% < 0.1% < 0.1% < 0.1% including manniotriosis < 0.5% < 0.1% < 0.1% < 0.1% including raffinose 0.7% < 0.1% 1.7% < 0.1% including stachyose 6.1% < 0.1% 5.6% < 0.1% dont verbascose 6.1% < 0.1% 4.4% < 0.1% Lipids 5.0% 8.4% 4.9% ND Ashes 9.0% 4.3 5.5% ND Degree of hydrolysis DH 7.4% 4.5% 4.9% ND Solubility at pH 4 according to Test A 48.8% 60% 15% ND Solubility at pH 7 according to Test A 52.2% 15% 34% ND Gel strength according to Test B 47

ND means not detected. When this refers to a measurement, ND means not measured.

The pea concentrate obtained according to the invention is particularly interesting because it contains approximately 60% protein on a dry basis and has a solubility at pH 4 or 7 of more than 45%.

It also has 12.9% GOS which will simply need to be defructosylated according to the teaching of example 2 to enrich it with this natural fiber.

The starch content of the pea concentrate obtained according to the invention (wet process) is lower compared to the tests “Pea protein concentrate marketed by LA-VITA under the name “pea protein concentrate” and VESTKORN pea protein concentrate obtained by dry process.

The combination of protein content and quality (globulins + albumins) and the presence of soluble natural fibers makes it a food ingredient of choice.

Example 2 : Reaction of defructosylation pea proteins:

We proceed as in example 1 but we carry out a defructosylation step on the protein fraction recovered in the overflow of the centrifugal decanter (Flottweg Z3) before the pH neutralization step.

The pH of the protein-enriched fraction thus obtained is rectified to 5.0 using hydrochloric acid and sodium hydroxide. The rectified protein-enriched fraction is heated and then temperature-controlled at 60°C. 0.2% of enzyme on dry weight of Sumizyme INV (invertase) is added. The reaction is left to react for 40 min with stirring.

The rest of the process is identical to example 1. The pH is then neutralized to 6.6. The solution is evaporated to 20% dry matter, undergoes HTST treatment at 130°C for 10 seconds with a flash at the outlet at 60°C. The solution is then atomized on a Nubilosa atomizer (air inlet temperature = 195°C – Air outlet temperature = 95°C). The pea protein powder named “Defructosylated pea concentrate according to example 2” was then analyzed.

Before
defructosylation after defructosylation Raffinose 0.7% < 0.1 Stachyose 6.1% < 0.1 Verbascose 6.1% 0.9% Manninotriosis < 0.5% 2.5% Melibiose < 0.1% 0.9% Fructose 0.20% 6.0%

GOS (in italics, about 12%) was hydrolyzed to defructosylated GOS (about 4.5%) and fructose (6%). Fructose provides a sweet flavor to soften the bitter aspect of pea proteins.

It should be noted that the enzymatic treatment using invertase can be carried out further downstream, i.e. on the concentrate obtained during step 1.

Claims (10)

Pea or field bean protein composition characterized in that it comprises a protein content of between 50% and 70% expressed in grams of protein per 100g of dry matter, said proteins consisting of a mixture of globulins and albumins, and between 3% and 15% of soluble fibers from pea or field bean galactooligosaccharides. Composition according to claim 2, characterized in that the dry weight ratio of globulins/albumins is between 70/30 and 90/10, preferably between 75/25 and 85/15. Composition according to one of claims 1 to 2, characterized in that the soluble fibers from pea or field bean galactooligosaccharides are selected from the list containing melibiose, manninotriose, verbascotetraose and their mixtures. Process for manufacturing a pea or field bean protein composition according to one of claims 1 to 3 comprising the following steps:
1. Preparation of an aqueous suspension of crushed pea or field bean seeds in an aqueous solution, said preparation being carried out, optionally, in the presence of a heat treatment;
2. Elimination of an insoluble fraction by solid/liquid separation of the aqueous suspension of crushed pea or field bean seeds obtained during step 1) allowing the production of a protein-enriched fraction;
3. Defructosylation of galactooligosaccharides from the protein-enriched fraction obtained during step 3) by enzymatic and/or fermentative means. Process according to claim 4, characterized in that the aqueous suspension of pea or field bean seeds from step 1 is obtained by adding a pea or field bean flour obtained by dry grinding prior to its dispersion in an aqueous solution. Method according to claim 4, characterized in that when the pea or field bean seeds are introduced in the form of whole pea or field bean seeds into the aqueous solution, step 1) of the method comprises a step of wet grinding of the aqueous composition formed between the pea or field bean seeds and the aqueous solution in order to obtain the aqueous suspension of ground pea or field bean seeds. Method according to one of claims 4 to 6, characterized in that the optional heat treatment of step 1 comprises:
(a) introducing pea or field bean seeds or crushed pea or field bean seeds into an aqueous solution at a temperature of between 65°C and 90°C in order to obtain an aqueous composition comprising pea or field bean seeds or crushed pea or field bean seeds;
b) heat treatment of the aqueous composition obtained in step a) formed between the pea or field bean seeds or the crushed pea or field bean seeds and the aqueous solution at a temperature between 40°C and 65°C for 1 to 10 min. Method according to one of claims 4 to 6, characterized in that the defructosylation of the galactooligosaccharide (GOS) of step 3 is carried out with an enzyme selected from invertase, alpha-galactosidae, beta-fructosidase and their possible combination. Method according to one of claims 4 to 6, characterized in that the defructosylation of galactooligosaccharide (GOS) is carried out with a microorganism of the genus Bacillus, preferably Bacillus subtilis, preferably a strain of Bacillus subtilis as deposited on May 28, 2020 at the CNCM under number I-5515. Use of said pea or field bean protein composition according to one of claims 1 to 3 or obtained according to one of claims 4 to 9 for the manufacture of food or beverage products, in particular plant-based alternatives to milk.