ENHANCED PEA OR BROAD BEAN PROTEIN

Abstract

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

Description

Title of the invention: IMPROVED PEA OR BROAD BEAN PROTEINS Scope of the invention

[0001] The invention relates to a new composition comprising pea or broad bean proteins combined with soluble fibers. Another object of the invention relates to a method for manufacturing these new pea protein compositions. The invention also relates to the use of said pea or broad bean protein compositions for the manufacture of food products. Previous art

[0002] 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).

[0003] In industrialized countries, protein intake is still predominantly in the form of animal protein. These proteins have good nutritional and functional properties that allow them to be used in a wide variety of food products.

[0004] However, numerous studies demonstrate that excessive consumption of animal protein at the expense of plant protein 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 in terms of the environmental impact linked to the harmful effects of intensive farming.

[0005] Thus, there is a growing demand from manufacturers for plant-based proteins with interesting nutritional and functional properties, without having the disadvantages of animal-based proteins.

[0006] Since the 1970s, the pea has been the most widely cultivated legume in Europe, and particularly in France, notably as a protein source for both animal and human consumption. Peas contain approximately 27% protein by weight. The term "pea" is used 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 their usual uses (human consumption, animal feed, and / or other). uses). Pea protein, mainly pea globulin, has been extracted and used industrially for many years.

[0007] The broad 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 family Fabaceae, subfamily Faboideae, tribe Fabeae. It is the same species as the broad bean, a plant used since antiquity for human consumption. The word broad bean then refers to both the seed and the plant.

[0008] We can first mention the so-called "dry" processes for extracting protein from peas or broad beans. The principle of these processes is to grind the seeds into a flour, which is then introduced into a turbo-separator, a device used to classify particles according to their size and density within an airflow. Turbo-separation makes it possible to obtain a protein-enriched fraction and a starch-enriched fraction. As will be explained later in this presentation, the protein-enriched fraction has a content of approximately 40%-60% and still contains between 2% and 15% starch. This starch is a polysaccharide fraction that is not necessarily desirable because it contributes to an increase in blood glucose levels; its replacement with polysaccharides that are indigestible by the consumer but digestible by their gut microbiota would be beneficial. These same properties are also observed in the case of broad beans.

[0009] Another example of a process for extracting protein from peas or broad beans is patent EPI400537, which describes so-called "wet" extraction processes. In this process, the seed is ground in the absence of water (a process known as "dry milling") to obtain a flour. This flour is then suspended in water at room temperature to carry out the various protein extraction steps. This type of process separates, by isoelectric precipitation, the proteins belonging to the globulin subgroup (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 galactooligosaccharides (GOS) from peas, which are not easily digested by humans, salts, 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 broad beans.

[0010] Several problems are easily foreseen, such as the proteins being separated into two fractions, requiring the use of two different fractions, or the GOS being combined with poorly digested albumins, necessitating post-treatment that complicates and increases the cost of the process. Finally, the massive presence of starch in the protein-enriched fraction of the concentrates obtained by Drying, by turbo-separation, is also a nutritional disadvantage for some formulations.

[0011] The Applicant has thus, after extensive research, developed a new manufacturing process for providing a composition containing all the so-called soluble proteins of peas or broad beans, that is, including all the globulins and albumins, as well as a quantity of soluble fiber derived from GOS, thus providing a high-quality nutritional source. Summary of the invention

[0012] Thus, the invention relates to a process for manufacturing pea or broad bean protein comprising the following steps: 1. Preparation of an aqueous suspension of crushed pea or broad 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 broad bean seeds obtained in step 1) allowing the obtaining of a protein-enriched fraction; 3. Defructosylation of galactooligosaccharides from the protein-enriched fraction by enzymatic and / or fermentative means.

[0013] Preferably, the aqueous suspension of crushed pea or broad bean seeds is obtained by adding a pea or broad bean flour obtained by dry grinding prior to its dispersion in the aqueous solution.

[0014] When pea or broad bean seeds are introduced in the form of whole pea or broad bean seeds into the aqueous solution, step 1 of the process includes a wet grinding step of the aqueous composition comprising the pea or broad bean seeds in order to obtain the aqueous suspension of ground pea or broad bean seeds.

[0015] 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 is understood to mean a temperature between 15°C and 27°C, preferably between 19°C and 23°C, the temperature values ​​being 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.

[0016] Preferably, the optional heat treatment in step 1 comprises: a) introducing pea or broad bean seeds or ground pea or broad bean seeds into an aqueous solution with a temperature between 65°C and 90°C in order to obtain an aqueous composition comprising ground pea or broad bean seeds; b) heat treatment of the aqueous composition obtained in step a) at a temperature between 40°C and 65°C for 1 to 10 min.

[0017] 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.

[0018] Preferably 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 filed on May 28, 2020 with the CNCM under number 1-5515.

[0019] Another object of the invention relates to pea or broad bean protein characterized in that it comprises a protein content of between 50% and 70% expressed in grams of protein on 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 galactooligosaccharides of pea.

[0020] Preferably, the dry weight globulin / albumin ratio is between 70 / 30 and 90 / 10, preferably between 75 / 25 and 85 / 15.

[0021] Preferably, soluble fibers from galactooligosaccharides of pea or broad bean are selected from the list containing melibiose, manninotriose, verbascotetraose and mixtures thereof.

[0022] Another object of the invention also relates to the use of said pea or broad 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

[0023] The invention relates to a process for manufacturing pea or broad bean protein comprising the following steps: 1. Preparation of an aqueous suspension of crushed pea or broad 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 broad bean seeds from step 1) allowing the obtaining of a protein fraction; 3. Defructosylation of galactooligosaccharides from the protein fraction by enzymatic and / or fermentative means.

[0024] Step 1) involves introducing peas or broad beans into an aqueous solution. The pea or broad bean seeds used in step 1) may have previously undergone steps well known to those skilled in the art, such as cleaning (removal of unwanted particles such as stones, dead insects, soil residues, etc.) or the removal of the external fibers of the pea or broad bean (cellulosic outer layer) by a well-known step called dehulling.

[0025] Thus, in step 1, "peas" or "broad beans" means whole peas or broad beans, or peas in the form of cotyledons, the outer husk of which has preferably been removed. Alternatively, ground pea seeds (i.e., pea flour) may be used, such ground peas generally being obtained by dry grinding.

[0026] The aqueous solution may be water which may optionally include additives such as, in particular, antifoaming compounds, salts or bacteriostatic agents.

[0027] The weight ratio of peas to aqueous solution in step 1) may in particular be between 0.1 and 2. In the embodiment where the pea or broad bean seed is ground into flour before being introduced into the aqueous solution, the weight ratio of peas to 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%.

[0028] According to one embodiment, the pH of the suspension in step 1) is adjusted to between 8 and 10. This adjustment can be made by adding a base such as sodium hydroxide, lime, or potassium hydroxide, preferably sodium hydroxide. According to another embodiment, the pH is not adjusted at this step.

[0029] The aqueous suspension of crushed pea or broad bean seeds is obtained in a step 1) by introducing into the aqueous solution, possibly preheated to a temperature between 65 °C and 90 °C if heat treatment is implemented, the crushed pea or broad bean seeds or the crushed pea or broad bean seeds.

[0030] In the case of heat treatment, the temperature of the aqueous solution can be pre-set to a temperature between 65°C and 90°C. Heating can be carried out using any equipment 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 around 75°C.

[0031] Alternatively, the aqueous solution undergoes no prior heating and is directly at a temperature between 65°C and 90°C upon mixing with the pea or broad bean seeds, or the ground pea or broad bean seeds. Preferably, the heat treatment temperature is between 40°C and 60°C, or even between 45°C and 55°C. Preferably, the heat treatment is carried out for 2 to 4 minutes.

[0032] According to either of these embodiments, the process then comprises heat-treating the suspension formed between the pea or broad bean seeds, or the ground pea or broad bean seeds, and the aqueous solution at a temperature between 40°C and 65°C for 1 to 10 minutes. The aqueous composition comprising pea or broad bean seeds, or ground pea or broad bean seeds, can be heated or cooled to reach this temperature.

[0033] In the case where pea or broad bean seeds (and not pre-ground pea or broad bean flour) are used in preparing the suspension according to step 1), the process includes a wet grinding step of the aqueous composition comprising pea or broad bean seeds or ground pea or broad bean seeds to obtain an aqueous suspension of ground pea or broad bean seeds. If heat treatment is applied, the wet grinding takes place after said heat treatment. Preferably, the process is carried out using pea or broad bean seeds, and the wet grinding step is performed by continuous passage through one or more mills to obtain the aqueous suspension of ground pea or broad bean seeds.The mill(s) can be any type of mill suitable for wet milling, such as wet ball mills, conical wet mills, helical wet mills, or wet mills equipped with rotor / stator systems. In one variant, the mill can be the one used in the examples in document WO2019 / 053387 on behalf of the Applicant. In the variant where the mill is of the rotor-stator type, this type of mill can allow continuous milling by passing the resulting aqueous composition between the pea or broad bean seeds and the aqueous solution through said mill. In a preferred sub-variant, the process combines two cutting stages (pre-cut and then cut) using different rotor-stator mills for each of these cuts.The pre-cutting and cutting can be carried out consecutively, or alternatively, the cutting can take place after pre-cutting and storage of the resulting aqueous composition between the pea or broad bean seeds and the treated aqueous solution. Such mills are described in document WO2019 / 158589. Optionally, dilution with water can be carried out during or at the end of this step to form the aqueous suspension of ground pea or broad bean seeds. In one variant, water is added continuously or intermittently during milling to dilute the aqueous composition. Generally, the dry matter content of the aqueous suspension of ground pea or broad bean seeds ranges from 10 to 30%, for example, from 15 to 25%.

[0034] Step 2) of the process consists of extracting the components from the aqueous suspension of ground pea or broad bean seeds, and in particular extracting a protein fraction by solid-liquid separation of a so-called insoluble fraction from the aqueous suspension of ground pea or broad bean seeds. According to one embodiment, Before carrying out the solid-liquid separation stage, it is possible to adjust the pH of the aqueous suspension of ground pea or broad bean seeds. The solid-liquid separation can then take place after adjusting the aqueous suspension of ground pea or broad bean seeds to a pH ranging from 6 to 9, preferably from 8 to 9, and most preferably from 8.5 to 9. This pH adjustment stage can be performed in a stirred tank. This stage can vary in length, lasting, for example, from 1 to 240 minutes, but generally from 5 to 60 minutes. To adjust the pH, any type of acid and / or base, organic or inorganic, or mixtures thereof, can be added. Examples of acids include hydrochloric acid, sulfuric acid, citric acid, or mixtures thereof. As a basic example, one can cite soda, potash or lime and their mixtures.The addition of base or acid, as well as the pH measurement, can be performed online. The base and / or acid can be in the form of aqueous solutions. Advantageously, before this solid-liquid separation, the aqueous suspension of ground pea or broad bean seeds is cooled to a temperature below 15°C. This temperature can range from 4 to 14°C, for example, from 10 to 12°C. This cooling step can be carried out using known techniques, such as passing the aqueous suspension of ground pea or broad bean seeds through a heat exchanger.

[0035] Generally, the protein fraction is the soluble part of the aqueous suspension of ground pea or broad bean seeds, and the starch- and fiber-rich fraction is the insoluble part. It is also possible to separate more than two insoluble fractions, for example, to recover a first insoluble fraction richer in starch and a second insoluble fraction richer in fiber. Thus, according to one variant of the process, a starch-rich fraction and / or a fiber-rich fraction is recovered from the insoluble part obtained from the solid-liquid separation step 2).

[0036] The term "starch-rich fraction and fiber-rich fraction," or "insoluble fraction," generally refers to a fraction comprising at least 50% starch and / or fiber. Methods for quantifying starch and fiber are known to those skilled in the art, and specific methods are described later. These fractions are recovered conventionally using known separation methods. Solid-liquid separation can be achieved, in particular, by means of at least one separation step with a decanter, specifically a decanter centrifuge, a centrifuge, or hydrocyclones. The process can also recover one or more fractions enriched in fiber and / or starch, which are removed from the suspension, and recover the protein fraction required for subsequent processing of the invention.

[0037] This step is key in differentiating it from concentrates obtained by dry separation. 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, as will be discussed later in this application.

[0038] Following step 2), the process may include a step of adjusting the pH of the pea or broad 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 then raised by adding a basic aqueous solution.

[0039] Step 3)

[0040] The process then includes a step 3) of defructosylation of galactooligosaccharides by enzymatic and / or fermentative means.

[0041] For the purposes of this invention, "galactooligosaccharides" means oligomers formed from a number n of sugars (monosaccharides) by alpha or beta glycosidic linkage and naturally present in peas or broad beans such as raffinose or stachyose.

[0042] Preferably, the water-soluble fraction according to the invention comprises defructosylated galactooligosaccharides selected from the list containing melibiose, manninotriose and verbascotetraose.

[0043] By "melibiose", in the context of the present invention, we mean the diholoside consisting of a galactose unit linked to a glucose unit by an α(1->6) glycosidic bond.

[0044] By "manninotriose", in the context of the present invention, we mean the triholoside consisting of the linkage of a galactose unit linked by a glycosidic bond α(1->6) to another galactose unit, itself linked to a glucose unit by another bond α(1->6).

[0045] By "verbascotetraose", also called "manninotetraose", in the context of the present invention, we mean the tetraholoside consisting of the chain of three galactose units linked by glycosidic bonds α(1->6), the third galactose unit being itself linked to a glucose unit by another α(1->6) bond.

[0046] 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 are preferred. Preferably, those skilled in the art will use the HPAEC-PAD amperometric assay method, and in particular with the following equipment: - Dionex Carbopac PA1 4*50mm pre-column - Ref. 43096 - Dionex Carbopac PA1 4*250mm column - Ref. 35391 - The detector is a PAD type, specifically a gold cell - The eluents are: Solvent A / 0.1M NaOH: Stir 4 liters of water under helium (flow rate: 100 ml / min) for 15 minutes. Add 20 ml of 50% NaOH. Restir under helium at 40 ml / min. Solvent B / 0.1M NaOH + 0.5M sodium acetate. Weigh 82g of sodium acetate directly into the container. Add 21ml of water. Stir and aerate under helium (flow rate: 100ml / min) for 15 minutes, then add 10ml of 50% NaOH, stir again, and aerate under helium. The helium flow rate can be reduced to 40ml / min.

[0047] Standards are used to calibrate HPLC and in particular: Reagents Reference Melibiose Fluka ref 63630 Raffinose Sigma ref R-0514 Stachyose Shna ref S-4001 Verbascose Fluka ref 56217

[0048] An internal standard is also used: Panose ref SIGMA P-2407 60mg in 100ml of water.

[0049] The injected volume is 5 µL at a temperature of 15°C. The analysis time is 90 min with a column temperature of 30°C and an injector sensitivity of 300 nC or 5 pA

[0050] 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

[0051] The oxidation program of the PAD detector is as follows: Time (min) Potential Integration 0 +0.05 0.20 +0.05 Start 0.40 +0.05 End 0.41 +0.75 0.60 +0.75 0.61 -0.15 1.0 -0.15

[0052] 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 Tl 10 5 5 5 T2 25 10 10 10 T3 50 15 15 15 T4 75 25 25 25 T5 100

[0053] Take 1 ml of control (from the 2 curves) + 1 ml Internal standard, qsp 20ml of water.

[0054] Weigh the quantity in mg of sample, add 1 ml of internal standard and adjust to 20 ml of water.

[0055] Filter on GxF / GHP 0.45 pm ref 4559T.

[0056] In a first alternative, the defructosylation of galactooligosaccharide (GOS) is carried out with an enzyme selected from invertase, alpha-galactosidase, beta-fructosidase and their possible combination.

[0057] Preferably, the enzyme used is an invertase such as Sumizyme INV. The pH and temperature are adjusted to optimize the defructosylation reaction, for example, pH 5 and 55°C for Sumizyme INV. The reaction is monitored by performing an analysis, for example, by HPLC, of ​​the sugars present in solution. When defructosylation is sufficient or even complete, it is neutralized using bases such as sodium hydroxide, followed by a heat treatment to inhibit the enzyme, for example, 130°C for 10 seconds.

[0058] In a second alternative, 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 filed on May 28, 2020 with the CNCM under number 1-5515.

[0059] According to the invention, "fermentation" refers to metabolic processes that generally convert carbohydrates into acids, gases, or alcohols to extract some of their chemical energy while reoxidizing the coenzymes reduced by these reactions. This is a redox metabolic pathway in which the ultimate electron acceptor is often the same as the final reaction product. It is characterized by partial degradation of the fermentable substance and allows only limited energy production. It occurs in yeasts and bacteria, as well as in muscle cells lacking oxygen, i.e., under anaerobic conditions.

[0060] In this process step, it is important to defructosylate the GOS without altering its carbon skeleton, without hydrolyzing it. Some prior art solutions address this technical problem. The present invention prefers to transform them into fibers beneficial for human and animal nutrition.

[0061] Steps 1), 2), and 3) of the process are carried out in that order. However, other optional steps may be implemented between steps 1), 2), and 3), such as pH adjustment, if deemed necessary by a person skilled in the art. Preferably after the defructosylation step, the process may include an additional heat treatment step 4) of the pea or fava bean protein. The temperature and time conditions may vary widely in this step, for example, from 70 to 140°C and 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 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 from 0.1 seconds to 5 minutes.According to another embodiment, 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 be intended to functionalize and / or sterilize the pea protein. To carry out this additional heat treatment step, the pea or fava bean protein may be in the form of an aqueous dispersion, preferably having a dry matter content ranging from 10 to 25%, for example from 15 to 20%. Advantageously, the process of the invention includes, following the additional heat treatment step, a cooling step of the pea protein. According to a preferred embodiment, this cooling step is achieved by 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 rapid cooling step ("flash-cooling") is achieved by applying a vacuum to the aqueous dispersion of pea protein, the applied vacuum being determined according to the chosen cooling temperature.

[0062] According to one variant of the process, it includes an optional step 5) of shearing the pea or broad bean protein, for example, by passing the aqueous protein dispersion through a high-pressure pump. As an example of a high-pressure pump, one could cite the high-pressure pumps marketed by Silverson, also known as high-shear mixers, for example, those in the UHS range. Preferably, the shearing step is carried out by a high-pressure pump.

[0063] The shearing step can take place before or after the heat treatment and / or pH raising steps.

[0064] According to another variant, the process alternatively includes an optional step 5) of homogenizing the pea protein.

[0065] To carry out this homogenization step, any type of homogenizer can be used. According to the invention, this refers to 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. Passing the aqueous dispersion of pea or broad bean protein through the homogenizer can thus enable the homogenization of the pea protein. The homogenization can be low-pressure, high-pressure, or ultra-high-pressure. The homogenization pressure can vary widely and, depending on the homogenization technique used, range from 1 to 1000 bar, for example, from 20 to 800 bar.In one version, the homogenization pressure ranges from 20 to 200 bar, for example, from 50 to 150 bar. In another version, the homogenization pressure ranges from 200 to 800 bar, for example, from 300 to 800 bar. In one version, the homogenization is single-effect. In another version, the homogenization is multi-effect, for example, double-effect. Suitable homogenizers are available from companies such as GEA or Tetra Pak.

[0066] The homogenization step can take place before or after the heat treatment and / or pH raising steps.

[0067] The process according to the invention may also include an optional step 6) of membrane nanofiltration, preferably carried out with a cutoff threshold between 150 Da and 300 Da. This step allows for pre-concentration of the composition before potential subsequent evaporation and / or drying steps, and also for desalting. Tests have shown that with a cutoff threshold of 300 Da, the ash content can be reduced from 8 to 10% initially to values ​​of 5 to 7%.

[0068] To measure the ash content, a person skilled in the art will use any method well known in the field. Preferably, a person skilled in the art will proceed as follows: - Weigh a sample weight PI - Place the sample for 24 hours in an oven at 550°C - Weigh the new sample weight P2 Ash content = (P2 / Pl)*100.

[0069] The process according to the invention may also include 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%, and 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, drum drying, or spray drying. The process may also include a grinding or micronization step. Spray drying is the preferred technology, in particular multi-effect spray drying. The pea or fava bean protein may be in powder form with a particle size d50, which may vary widely, for example from 10 to 500 µm, generally from 50 to 150 µm.

[0070] By "d50" in the present invention, we mean 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.

[0071] To perform this d50 measurement, a laser particle size analyzer is preferably used, and 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.

[0072] Preferably, drying step 7) may comprise a first substep, a concentration step by evaporation, followed by the actual drying step. This evaporation is conventionally carried out using any suitable 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 both eliminate undesirable volatile compounds and optimize the drying step.

[0073] 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.

[0074] Pea or broad bean protein

[0075] Another object of the invention relates to a pea or broad bean protein composition that can be obtained by the process of the invention.

[0076] Another object of the invention relates to the composition of pea or broad bean protein characterized in that it comprises a protein content of between 50% and 70% expressed in grams of protein on 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 galactooligosaccharides of pea.

[0077] By "pea protein composition" is meant a composition which comprises principally, but not exclusively, pea proteins. Such a composition may contain residual impurities such as, for example, minerals, sugars, etc.

[0078] By "fava bean protein composition" is meant a composition which comprises principally, but not exclusively, fava bean proteins. Such a composition may contain residual impurities such as, for example, minerals, sugars, etc.

[0079] The term “pea” in this application shall be understood as all wild varieties of “smooth pea” and all mutant varieties of “smooth pea” and “wrinkled pea.” The term “pea” is to be understood here 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 to which these varieties are generally put (human food, animal feed, and / or other uses). The term “pea” in this application includes pea varieties belonging to the genus Pisum and more particularly to the species sativum and aestivum.These mutant varieties include those designated "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.

[0080] The term "protein" in this application should be understood as macromolecules formed from one or more polypeptide chains consisting of a chain of amino acid residues linked together by peptide bonds. In the specific 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).

[0081] For the purposes of this invention, "globulins" refers to proteins soluble in neutral saline solutions. Pea or broad bean globulins are mainly subdivided into three subfamilies: legumins, vicilins, and convicilins.

[0082] For the purposes of this invention, "albumin" refers to proteins soluble in pure water. Pea albumins, present in pea or broad bean proteins at a level of approximately 20%, are mainly subdivided into two families designated PA1 and PA2.

[0083] Generally, the protein content by weight of the pea or broad 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%, including 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, or 70%.

[0084] The pea or broad bean protein according to the invention obviously generally includes other minor constituents other than proteins, such as starch, lipids, fibers, and / or sugars.

[0085] Generally, the total starch content in the pea or broad bean protein produced according to the method of the invention is between 0% and 0.5%, for example from 0.1% to 0.4%, in particular from 0.2% to 0.3%. This total starch content can be measured using method AO AC 996.11. The residual starch contents can therefore be 0%, 0.1%, 0.2%, 0.3%, 0.4%, or 0.5%. This particularly low starch content is characteristic of the pea or broad bean protein according to the invention. Pea or broad bean protein concentrates obtained by turboseparation contain more starch, for example between 2 and 5% according to the Feedipedia webpage (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 its functional properties, such as increased viscosity when heated.

[0086] Generally, the total lipid content ranges from 0 to 15%, for example from 1 to 10%. The total lipid content can be determined by the AO AC 996.06 method in acid hydrolysis.

[0087] Preferably, the dry weight globulin / albumin ratio is between 70 / 30 and 90 / 10, preferably between 75 / 25 and 85 / 15.

[0088] Preferably, the pea or broad 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%.

[0089] In the present invention, the term "degree of hydrolysis" refers to the percentage ratio between the quantity of amine (or carboxylic) groups of free amino acids and the total quantity, including both free groups and those involved in a peptide bond (a chemical bond characteristic of proteins resulting from the association of a carboxylic group of one amino acid and an amine group of another). For a protein composition consisting of all its amino acids linked together, and therefore having only one free amine and one free carboxylic group, this degree of hydrolysis will be 0%. Conversely, for a protein composition in which all the same amino acids are said to be "free," that is, in which both their amine and carboxylic groups are not involved in peptide bonds, this degree of hydrolysis will be 100%.

[0090] Several methods exist for quantifying the degree of hydrolysis. They all consist primarily of the colorimetric determination of free amine (or carboxylic) groups, followed by hydrolysis to break all peptide bonds, and finally, a colorimetric determination of the total amine (or carboxylic) groups. The calculated percentage of free amines (or carboxylics) relative to the total amines gives the degree of hydrolysis. Any well-known method can be used, such as the TNBS method or the OPA method. In the present invention, the OPA method is preferred, and a measurement procedure is described below:

[0091] The amino nitrogen (free NH2) content of the protein sample according to the invention is first determined using the MEGAZYME kit (reference K-PANOPA). The protein nitrogen (total nitrogen) content of the sample is also determined. The degree of hydrolysis can then be calculated.

[0092] Determination of amino nitrogen content:

[0093] 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.

[0094] The amount of isoindole derivative formed during this reaction is stoichiometric with respect to the amount of free amino nitrogen. It is the isoindole derivative that is measured by the increase in absorbance at 340 nm.

[0095] In a 100 mL beaker, an exactly weighed test portion P* of the sample to be analyzed is introduced. This test portion will be from 0.5 to 5.0 g depending on the amino nitrogen content of the sample. Approximately 50 mL of distilled water is added, the mixture is homogenized, and the solution is transferred to a 100 mL volumetric flask. 5 mL of 20% sodium dodecyl sulfate (SDS) is added, and the volume is brought to 100 mL with distilled water. The solution is stirred for 15 minutes with a magnetic stirrer at 1000 rpm. Solution No. 1 is prepared by dissolving one tablet of the Dissolve one Megazyme tablet (vial 1) in 3 mL of distilled water and shake until completely dissolved. One tablet is required per test. Solution 1 is prepared immediately before use.

[0096] A blank, a standard and a sample are prepared directly in the spectrophotometer cuvettes under the following conditions: - white: introduce 3.00 ml of solution no. 1 and 50 ml of distilled water - standard: introduce 3.00 ml of solution no. 1 and 50 pl from vial 3 of the Megazyme kit - sample: introduce 3.00 ml of solution no. 1 and 50 pl of the sample preparation.

[0097] The contents of each cuvette are mixed and the absorbance (Al) measurement of the solutions is read after approximately 2 minutes using a spectrophotometer at 340 nm (spectrophotometer equipped with cuvettes of 1.0 cm optical path, capable of measuring at a wavelength of 340 nm, and verified according to the operating procedure described in the manufacturer's relevant technical manual).

[0098] The reactions are then initiated immediately by adding 100 pl of solution no. 2, which corresponds to the OPA solution from bottle 2 of the Megazyme kit, into each spectrophotometer cuvette.

[0099] The contents of each tank are mixed and placed in the dark for approximately 20 minutes.

[0100] The absorbance measurement A2 of the blank, standard and sample is then read by spectrophotometer at 340 nm.

[0101] The content of free amino nitrogen, expressed as a percentage by weight relative to the weight of the product, is given by the following formula: % amino nitrogen = (àAech- AÆMc)x3.15x14.0 IXVxlOO % amino nitrogen = 6803 x 0.05 xm X1000 (AAech- AAbIc)xl2.974x V MX1000 Or : AAech = Aech2 - Aechl AAblc = Ablc2 - Ablcl Aech2 = absorbance of the sample after the addition of solution #2 Aechl = absorbance of the sample after addition of solution n°1 Ablc2 = absorbance of the blank after addition of solution #2 Ablcl = absorbance of the blank after addition of solution no. 1 V = volume of the flask m = mass of the test sample in g 6803 = extinction coefficient of the isoindole derivative at 340 nm (in L.mol'.cm*). 14.01 = molar mass of nitrogen (in g.mol1) 3.15 = final volume in the tank (in mL) 0.05 = test portion in the tank (in mL)

[0102] Determination of protein nitrogen content:

[0103] 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.

[0104] Calculation of the degree of hydrolysis

[0105] The degree of hydrolysis (DH) is calculated using the following formula: ν _ % amino nitrogen vmn 27 / 1 — % protein nitrogen X iUU

[0106] The pea or broad bean protein according to the invention contains between 3% and 15% soluble fiber, preferably between 4% and 8%, derived from pea galactooligosaccharides. The content can therefore be between 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% and 15%.

[0107] Preferably, soluble fibers from galactooligosaccharides of pea or broad bean are selected from the list containing melibiose, manninotriose, verbascotetraose and mixtures thereof.

[0108] The combination of the protein, composed of a mixture of globulin and albumin, and defructosylated soluble fibers allows for the production of a single ingredient of significant interest. With this single ingredient, it is possible to provide both a high-quality protein and a quantity of soluble fiber. 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.

[0109] The properties of pea or broad bean protein can vary widely, depending on the process parameters set out above and as shown in the Examples section.

[0110] 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 be in any intermediate range (i.e., 11%, 12%, 13%... 97%, 98%, 99%), and those skilled in the art will know, based on the process indications given above and in the Examples section, how to modify the process parameters within the specified ranges to achieve the desired solubility. Advantageously, the dried pea or fava bean protein has a solubility at pH 7.0 and pH 4.0 ranging from 40% to 55%.

[0111] Solubility: Test A

[0112] As regards solubility, it is determined according to the TEST A method described below: Measurement of solubility in water This measurement is based on diluting the sample in distilled water, centrifuging it, and analyzing the supernatant.

[0113] Operating procedure: In a 400 ml beaker, introduce 150 g of distilled water at a temperature of 20°C + / - 2°C, mix with a magnetic stir bar and add precisely 5 g of the sample to be tested. Adjust the pH to the desired value with NaOH or 0.1 N HCl (pH 7). Top up the water content to 200g. Mix for 30 minutes at 1000 rpm and centrifuge for 15 minutes at 3000 g. Collect 25 g of the supernatant. Place in a crystallizing dish that has been previously dried and weighed. 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.

[0114] The soluble solids content, expressed as a percentage by weight, is given by the following formula:

[0115] [Math. 1] (ml ~ m2) x (200 * W) x 100 .........................................= % saiiOty PI KP Or : P = weight, in g, of the sample = 5 g ml = weight, in g, of the crystallizing dish after drying m2 = weight, in g, of the empty crystallizing dish PI = weight, in g, of the collected sample = 25 g

[0116] According to one embodiment, dried pea or broad bean protein may have a gelling capacity. This gelling capacity may range from 1 to 100 Pa, preferably between 10 and 90 Pa, and even more preferably between 30 and 60 Pa.

[0117] Gelling power: Test B

[0118] By "gelling power" is meant the functional property consisting of the ability of a protein composition to form a gel or network, thereby increasing viscosity and generating an intermediate state of matter between liquid states and solids. The term "gelling strength" can also be used. To quantify this gelling power, it is therefore necessary to generate this network and evaluate its strength. To perform 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 containing 15% + / - 2% dry matter and at pH 7; 2) Agitation 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 a constraint rheometer equipped with a concentric cylinder; 5) Measurement of the 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 minutes and measurement of G'2 after stabilization at 20°C + / - 2°C #; 6) Calculation of the gelling power equal to G'2 - G'1.

[0119] Preferably, the constraint rheometers are selected from the DHR 2 (TA, instruments) and MCR 301 (Anton Paar) models, with a concentric cylinder-type spindle. They have a Peltier temperature control system. To avoid evaporation problems at high temperatures, paraffin oil is added to the samples.

[0120] A “#rheometer#” as defined in 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 dimensions (very low mechanical inertia of the rotor), it allows for the fundamental study of the mechanical properties of a liquid, gel, suspension, paste, etc., in response to an applied force.

[0121] So-called "imposed stress" models allow, by applying a sinusoidal load (oscillation mode), the determination of the intrinsic viscoelastic quantities of the material, which depend in particular on time (or angular velocity φ) and temperature. In particular, this type of rheometer allows access to the complex modulus G*, which itself allows access to the moduli G' or elastic part and G" or viscous part;

[0122] The first three steps consist of resuspending the protein in water, under precise conditions to maximize the subsequent measurement.

[0123] The water chosen is preferably reverse osmosis water, but it is also possible to use drinking water.

[0124] Its temperature is 60°C + / - 2°C during the initial resuspension (steps 1 and 2) and then 20°C + / - 2°C after solubilization for 24 hours and cooling before measurement (step 3). Generally, 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.

[0125] A defined quantity of protein is added to said water to obtain a suspension with a dry matter content of 15% + / - 2%. This is done using equipment well known to those skilled in the art, such as beakers and magnetic stir bars. A volume of 50 mL is stirred for a minimum of 10 minutes at 350 rpm at room temperature. Generally, and unless otherwise specified, the dry matter contents given in this 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.

[0126] The fourth step consists of introducing the sample into the rheometer by covering it with a thin layer of oil in order to limit evaporation.

[0127] During the fifth step, the following temperature schedule is then applied#: 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 minutes.

[0128] The measurement of the parameter G' is carried out continuously during this scale and is recorded.

[0129] The sixth and final step of test B consists of processing 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.

[0130] The gelling power is equal to G'2 - G'1.

[0131] According to an optional embodiment, the pea or broad bean protein is an enzymatically modified protein. By enzymatically modified protein, a person skilled in the art understands a protein whose protein structure has been intentionally modified by the addition to the protein of at least one enzyme capable of modifying the protein structure. This enzyme may be selected from proteases, peptidases, deamidating enzymes, for example those of type EC 3.5.1 such as glutaminase, or deiminating enzymes, for example those of type EC 3.5.3 such as peptidylarginine deiminase. These protein-modifying 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 reduces astringency, particularly the astringency of rapeseed protein. If the process includes proteolysis of the protein-enriched fraction, this can 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%. Those skilled in the art will be able to adapt the conditions of enzymatic proteolysis, or even omit this step altogether, to obtain the desired DH. According to a preferred embodiment of the invention, the pea or broad bean protein is not enzymatically modified by deamination. According to another preferred embodiment of the invention, the protein is not enzymatically modified.One advantage of the invention is that it is possible to modify the organoleptic properties of pea protein, and in particular to give it a milky aromatic profile, without even needing to enzymatically modify the protein. The invention therefore makes it possible to supply, according to one embodiment, proteins with an unmodified primary structure.

[0132] Utilization of pea protein

[0133] The invention also relates to the use of the pea or broad bean protein of the invention for the manufacture of food or beverage products, in particular plant-based alternatives to milk.

[0134] Generally, the pea or broad bean protein of the invention can be used in food and beverage products that 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 ranging from about 1% by weight to about 80% by weight relative to the total dry weight of the food or beverage product. All intermediate amounts (i.e., 2%, 3%, 4%... 77%, 78%, 79% by weight relative to the total weight of the food or beverage product) can be used, as well as all intermediate ranges based on these amounts. These food and beverage products can be adapted for vegetarian or vegan populations.

[0135] A particularly interesting use of the protein of the invention relates to its use in beverages that have a more pleasant taste than those obtained from other commercially available pea proteins. The pea or broad bean protein of the invention can advantageously be used for the manufacture of beverages, in particular milk alternatives, or in other words, milk substitutes. Moreover, due to the milky aromatic note imparted by the ingredient, these beverages can also have a more milky aromatic note than a beverage not containing said protein, which is an undeniable advantage for the manufacture of plant-based milk alternatives. In addition to an improvement in aroma, it is also possible according to the invention of obtaining a more coating texture in the mouth (mouthfeel effect) than when other pea or broad bean proteins are used, which is advantageous for drinks, and especially for plant-based alternatives to milk because animal milks generally also have a coating texture.

[0136] In beverages, the protein content of these products can vary widely and can also be a high-protein drink. The protein content can range, for example, from 1 to 12% by dry weight relative to the total weight of the beverage, and in particular from 3 to 10% relative to the total weight of the beverage. Beverages can be of any type and include plant-based alternatives to milk or milk substitutes, including barista-style milks or coffee creamers.This can also include other ready-to-drink beverages, acidic or not, such as carbonated drinks (including, but not limited to, carbonated soft drinks), non-carbonated drinks (including, but not limited to, non-carbonated soft drinks such as flavored waters, fruit juices, and sweetened or unsweetened tea or coffee-based drinks), alcoholic beverages such as beers or spirits, smoothies, and beverage concentrates (including, but not limited to, liquid concentrates and syrups, as well as non-liquid "concentrates," such as freeze-dried and / or powdered preparations or "powder mixes"). It should be noted that in beverages, flavorings or masking agents are generally used to reduce the pea or broad bean flavor, 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 the amount of flavoring or masking agents, or even to completely eliminate these constituents from the beverage, while still maintaining a very satisfactory taste. The beverages may also contain hydrocolloids; however, since the pea or fava bean protein provides a more coating texture, it is possible to reduce or even eliminate the hydrocolloid content while maintaining a coating texture in the mouth.

[0137] Food products that may be concerned include bakery products such as bread products (including, but not limited to, leavened and unleavened breads, sandwich breads, yeast breads and unleavened breads such as baking soda breads), breads comprising all types of wheat flour, breads comprising all types of flour other than wheat (such as potato, rice, barley, spelt and rye flours), gluten-free breads; mixes for the preparation of said bread products; sweet bakery products (including, but not limited to, rolls, cakes, pies, pastries, waffles, pancakes, muffins, pancakes, and biscuits); mixes for the preparation of said 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 and other products, such as cream fillings); snack bars (including, but not limited to, energy, cereal, nut, and / or fruit bars).

[0138] This may also include gelled desserts such as dessert creams or flans and puddings. Another type of dessert may 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 the like).

[0139] Other products conventionally prepared from animal milk may also include the pea or broad bean protein of the invention to form substitutes. These may be acidified and / or fermented products, for example, lactic, vegan, or mesophilic cultures. They may include yogurts (including, but not limited to, full-fat, reduced-fat, and fat-free yogurts, which may be free of milk proteins and lactose). The term "yogurts" also includes soft cheeses and fromage frais. They may also include cheese substitutes such as spreadable, processed, cooked and uncooked pressed cheeses, soft cheeses, stretched-curd cheeses, and blue cheeses; It can be Emmental, string cheese, ricotta, provolone, parmesan, munster, mozzarella, monterey jack, manchego, bleu, fontina, feta, edam, double Gloucester, camembert, cheddar, brie, asiago and Havarti.It can also include other products such as vegetable butters or fresh cream.

[0140] Other products that may include the pea or broad bean protein of the invention are also sauces such as salad dressings or sauces based on mayonnaise or ketchup or syrups.

[0141] Also, the pea or broad 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 the preparation of breakfast cereals. They can also be used in sweet spreads (including, but not limited to, jellies, jams, nut butters such as peanut butter, spreads, and other spreadable products).

[0142] The pea or broad bean proteins of the invention can also be used as a carrier or encapsulation of aroma.

[0143] Other types of food and beverages not mentioned here but which conventionally contain one or more proteins may also be considered within the scope of the present invention. In particular, animal feed (such as pet food) is explicitly considered.

[0144] Pea or broad bean protein can also be used, possibly after texturizing, in meat substitutes such as emulsified sausages or hamburgers, or in 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 refer to proteins textured by extrusion, i.e., in particular, dry extrusion (also known as Textured Vegetable Protein) or high-moisture extrusion. Extruders can be single-screw, twin-screw, or multi-screw. In the case of twin-screw extrusion, the extrusion can be co-rotating or counter-rotating. Examples of multi-screw extrusion include the planetary extruder and the ring extruder.Other more specific technologies could also be mentioned, such as shear cell technology, microextrusion, or 3D printing.

[0145] Food products or beverages may be used in specialized nutrition, for example for specific populations, such as babies or infants, children, adolescents, adults, the elderly, athletes, or people suffering from an illness. These may include nutritional meal replacement formulas, complete nutritional drinks, for example for weight management, or in clinical nutrition (e.g., tube feeding or enteral nutrition).

[0146] Pea or broad bean protein can be used as the sole source of protein, but can also be used in combination with other additional proteins, whether plant or animal. These additional proteins may 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 generally have a protein content between 50% and 70%, while isolates have a protein content greater than 70%, preferably between 80% and 90%, respectively. The term "plant protein" refers to all proteins derived from cereals, oilseeds, legumes, and tuberous plants, as well as all proteins derived from algae and microalgae or from Mushrooms, used alone or in mixtures, selected from the same family or from different families. The term "legume" generally refers to the family of dicotyledonous plants in the order Fabales. Several legumes are important cultivated plants, including soybeans, beans (particularly mung beans), chickpeas, broad beans, peanuts, cultivated lentils, cultivated alfalfa, various clovers, broad beans, carob, licorice, and lupins. The additional legume protein may be selected from these legumes or may be a pea or broad bean protein other than that of the invention. In this application, the term "cereals" refers to cultivated plants of the grass family that produce edible grains, such as wheat, oats, rye, barley, maize, sorghum, or rice. The tubers can be carrot, cassava, konjac, potato, Jerusalem artichoke, sweet potato.Oilseed plants are generally plants that produce seeds from which oil is extracted. Examples of oilseed plants include sunflower, rapeseed, peanut, sesame, pumpkin, and flax. Animal proteins can be, for example, egg or milk proteins, such as whey protein, casein, or caseinates. The pea or broad bean protein composition of the invention can thus be used in combination with one or more of these proteins or amino acids to improve the nutritional properties of the final product, for example, to enhance the PDCAAS of the protein or to provide other functionalities.

[0147] Pea or broad 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.

[0148] 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 to the present invention. Examples

[0149] Example 1: Pea protein concentrate

[0150] Approximately 1000 kg of peas were used. The outer fibers of the peas were first separated from the seeds by crushing (mechanical separation of the outer husk and the pea seed) and hulling (sorting the outer husks and hulled pea seeds using compressed air). The prepared seeds 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 resulting flour was mixed with water to obtain a suspension of ground pea seed flour with 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).

[0151] 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 using a Nubilosa atomizer (air inlet temperature = 195°C - air outlet temperature = 95°C). The pea protein powder, designated "Pea Concentrate according to Example 1," was then analyzed.

[0152] [Tables 1] Pea concentrate according to example 1 NUTRALYS® F85M (Pea protein isolate) Pea protein concentrate VESTK ORN Pea protein concentrate marketed by LA-VITA under the name "pea protein concentrate" Protein 62.0% 85.10% 54.0% 50-55% Carbohydrate 24.0% 4.2% 30.6% 12-15% of which starch 0.1% 0.1% 5.4% 12-15% of which glucose 1.6% <0.1% <0.1% <0.1% of which fructose 0.2% <0.1% <0.1% <0.1% of which sucrose 7.8% <0.1% 2.7% <0.1% of which melibiose <0.1% <0.1% <0.1% <0.1% of which manniotriose <0.5% <0.1% <0.1% <0.1% of which raffinose 0.7% <0.1% 1.7% <0.1% of which stachyose 6.1% <0.1% 5.6% <0.1% of which verbascose 6.1% <0.1% 4.4% <0.1% Lipids 5.0% 8.4% 4.9% ND Ash 9.0% 4.3 5.5% ND Degree of hydrolysis DH 7.4% 4.5% 4.9% ND Solubility at pH 4 salt on Test A 48.8% 60% 15% ND Solubility at pH 7 salt on Test A 52.2% 15% 34% ND Gel strength according to Test B 47

[0153] ND means not detected. When this designation refers to a measurement, ND means not measured.

[0154] 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%.

[0155] It also contains 12.9% GOS which will simply need to be defructosylated according to the teaching of example 2 to enrich it with this natural fiber.

[0156] The starch content of the pea concentrate obtained according to the invention (wet method) is lower compared to the tests "Pea protein concentrate marketed by LA-VIT A under the name "pea protein concentrate" and VESTKORN pea protein concentrate obtained by dry method.

[0157] The combination of the protein content and quality (globulins + albumins) as well as the presence of soluble natural fibers makes it a choice food ingredient.

[0158] Example 2: Defructosylation reaction of pea proteins:

[0159] The procedure is as in example 1 but a defructosylation step is carried out on the protein fraction recovered in the overflow of the centrifugal decanter (Flottweg Z3) prior to the pH neutralization step.

[0160] 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 the enzyme Sumizyme INV (invertase) by dry weight is added. The reaction is allowed to proceed for 40 minutes under stirring.

[0161] The remainder 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 using 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.

[0162] [Tables3] before defructosylation after defructosylation Raffinose 0.7% < 0.1 Stachyose 6.1% < 0.1 Verbascose 6.1% 0.9% Manninotriose < 0.5% 2.5% Melibiose <0.1% 0.9% Fructose 0.20% 6.0%

[0163] The GOS (in italics, approximately 12%) was hydrolyzed into defructosylated GOS (approximately 4.5%) and fructose (6%). The fructose provides a sweet flavor that helps to soften the bitterness of the pea proteins.

[0164] 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

Demands

1. Pea or broad bean protein composition characterized in that it comprises a protein content of between 50% and 70% expressed in grams of protein on 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 galactooligosaccharides of pea or broad bean.

2. Composition according to claim 1 characterized in that the dry weight ratio of globulins / albumins is between 70 / 30 and 90 / 10, preferably between 75 / 25 and 85 / 15.

3. Composition according to any one of claims 1 to 2 characterized in that the soluble fibers from galactooligosaccharides of pea or broad bean are selected from the list containing melibiose, manninotriosc.lc verbascotetraose and mixtures thereof.

4. A method for manufacturing a pea or broad bean protein composition according to any one of claims 1 to 3 comprising the following steps:

1. Preparation of an aqueous suspension of ground pea or broad 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 ground pea or broad bean seeds obtained in step 1) allowing the obtaining of a protein-enriched fraction; 3. Defructosylation of the galactooligosaccharides of the protein-enriched fraction obtained in step 3) by enzymatic and / or fermentative means.

5. The process according to claim 4 characterized in that the aqueous suspension of pea or broad bean seeds from step 1 is obtained by adding a pea or broad bean flour obtained by dry grinding prior to its dispersion in an aqueous solution.

6. The process according to claim 4, characterized in that when pea or broad bean seeds are introduced as whole pea or broad bean seeds into the aqueous solution, step 1) of the process comprises a wet grinding step of the aqueous composition formed between the pea or broad bean seeds and the solution aqueous in order to obtain the aqueous suspension of crushed pea or broad bean seeds.

7. A process according to any one of claims 4 to 6 characterized in that the optional heat treatment of step 1 comprises: a) introducing pea seeds or broad beans or ground pea seeds or broad beans into an aqueous solution having a temperature between 65°C and 90°C in order to obtain an aqueous composition comprising pea seeds or broad beans or ground pea seeds or broad beans; b) heat treatment of the aqueous composition obtained in step a) formed between the pea seeds or broad beans or ground pea seeds or broad beans and the aqueous solution at a temperature between 40°C and 65°C for 1 to 10 min.

8. A method according to any one of claims 4 to 6 characterized in that 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.

9. A process according to any 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 filed on May 28, 2020 with the CNCM under number 1-5515.

10. Use of said pea or broad bean protein composition according to any one of claims 1 to 3 or obtained according to any one of claims 4 to 9 for the manufacture of food or beverage products, in particular plant-based alternatives to milk.