Temperature-gelling pea proteins

Abstract

The invention relates to a functionalized pea protein having a gelling power, according to a TEST A, of at least 300 Pa, preferably in the range of 500 to 2000 Pa, most preferably in the range of 600 to 1800 Pa, and a determined viscosity of less than 0.65 Pa.s, the viscosity being measured at 15% by weight of dry matter, at a shear rate of 40s-1 and a temperature of 20°C, a method for obtaining such a protein, and the use thereof in food products or beverages.

Description

Description Title: GELLING PEA PROTEINS AT TEMPERATURE Field of invention

[0001] The present invention relates to a temperature-gelling pea protein which also has a reduced viscosity. A second subject of the invention relates to a method for manufacturing the pea protein. A third subject relates to the use of said pea protein in food applications. Prior art

[0002] Daily protein requirements are generally between 12 and 20% of the diet. 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 show that excessive consumption of animal proteins at the expense of plant proteins is one of the causes of increased 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.

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

[0006] Since the 1970s, the pea has been the most widely grown grain legume in Europe, and mainly in France, particularly as a protein resource for animal and human food. Peas contain approximately 27% protein by weight. The term "pea" is here considered 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 used industrially for many years. An example of a pea protein extraction process is patent EP1400537.In this process, the seed is ground in the absence of water (known as "dry grinding") to obtain a flour. This flour is then suspended in. water at room temperature in order to then proceed with the various stages of protein extraction.

[0007] Despite its undeniable qualities, pea protein suffers, compared to other plant proteins, from a lower gelling power at temperature than other plant proteins. For example, soy protein isolates have excellent gelling power at temperature. However, for many finished products (textured proteins by dry or wet extrusion, meat substitutes such as emulsified sausages, cold meat substitutes, egg substitute compositions or even dessert creams), the use of temperature-gelling proteins makes it possible to provide food products with an improved texture.

[0008] Since it is generally accepted that preserving the native structure of the protein allows its functionality to be preserved, processes for producing proteins with improved temperature gelling power generally involve steps following which the native character of the protein is maintained. Indeed, this is favorable to the development of gelling power when used in the final food product. Thus, document WO2022 / 174339 describes a method for treating legume proteins, in particular undenatured pea protein concentrates and isolates, to induce gelation under heat and in saline solution. The method includes maintaining at an alkaline or acidic pH before neutralization and / or cold atmospheric plasma treatment. Document WO2021 / 260169 also describes a protein with improved temperature gelling power.This protein is obtained by a process comprising an acid wash as well as an alkaline wash.

[0009] However, other methods for providing proteins with improved temperature gelling power, during which the proteins are denatured, have already been described. For example, WO2023 / 076541 describes the formation of gels using transglutaminase in an aqueous protein composition. However, the document does not describe the provision of protein with improved gelling power and reduced viscosity: the method modifies the structure of the protein by crosslinking it (and crosslinking increases the viscosity of proteins), does not allow the provision of protein with improved gelling power in dry form, let alone in the form of ready-to-use protein powder. Furthermore, the use of transglutaminase leads to a significant modification of the primary structure of the protein.Document WO 2020 / 221978 describes the manufacture of pea protein powder with improved gelling power in which a micronization step is implemented. However, the gelling power may still be insufficient and this document remains silent regarding the viscosity of the product obtained. However, it has been observed by the inventors that a micronized protein powder has a higher viscosity than that of non-micronized powder. Document WO2011 / 124862 describes a process for manufacturing soluble and functional pea proteins comprising a heat treatment step. from 0.01 to 1s at a temperature of 100 to 160°C followed by a cooling step. As demonstrated below in the Examples section of this Application, although heat treatment makes it possible to provide proteins with gelling power at temperature, this remains fairly insufficient.

[0010] With regard to isoelectrically precipitated plant protein concentrates, a process leading to a protein with improved temperature gelling power has been described in EP522800 A2. This process comprises the steps of suspending the protein at an alkaline pH at a temperature of 75-95°C for 1-120 minutes before neutralization and then spray drying. A problem with this process is that it simultaneously causes a significant increase in the viscosity of the product (a 17% dry matter suspension of the product thus treated has a viscosity five times higher than a suspension of the product before treatment). Thus, while there are methods for improving the gelling power of certain proteins, this improvement is accompanied by a significant modification of other properties and in particular a significant increase in its viscosity.In the Examples section, commercial pea proteins with improved gelling power at temperature are also reported: all of these pea proteins also have a much higher viscosity than pea proteins with reduced gelling power.

[0011] It follows from the above that there is a need for such proteins which, in addition to their gelling power at temperature, have a limited viscosity.

[0012] The Applicant has thus arrived, after much research, at a new manufacturing process making it possible to provide pea proteins with a high gelling power, but also a reduced viscosity. Thus, unlike the pea proteins already described, it is possible to provide fluid liquid compositions using these proteins, even when the concentration of these proteins is high. These proteins are also capable of forming a gel when heated and are therefore very interesting for the manufacture of the aforementioned finished products. Summary of the invention

[0013] The subject of the invention is thus a functionalized pea protein having a gelling power according to a TEST A of at least 300 Pa, preferably ranging from 500 to 2000 Pa, most preferably ranging from 600 to 1800 Pa and a determined viscosity of less than 0.65 Pa.s, the viscosity being measured at 15% by weight of dry matter, at a shear rate of 40s-1 and a temperature of 20°C.

[0014] The invention also relates to a method for manufacturing the functionalized pea protein of the invention which comprises: - a supply of an aqueous pea protein composition comprising pea protein precipitated at isoelectric pH and pea polypeptides, said polypeptides being insoluble at a pH below 8.5, the pea proteins of the aqueous composition having a percentage of denaturation of less than 65%, - adjustment of the aqueous composition to a pH ranging from 6.0 to 6.8, - heat treatment of the mixture to obtain the functionalized pea protein.

[0015] The protein manufacturing process developed by the inventors thus made it possible to obtain a functionalized pea protein combining both gelling power at temperature and, unlike previously known gelling pea proteins, reduced viscosity. To achieve this functionalized pea protein, the inventors used a process involving heat treatment of pea proteins previously placed in a restricted pH range of 6.0 to 6.8. As shown in the examples section, in this restricted range, it is possible for the functionalized pea protein to develop gelling power at temperature, while maintaining the desired viscosity.

[0016] As for the provision of the aqueous pea protein composition subjected to heat treatment, it uses a method for extracting polypeptides insoluble at a pH lower than 8.5 (i.e. proteins which, for example, are not present when extracting a flour in aqueous solution at the native pH of the flour). This extraction can for example be carried out by placing a pea material (pea flour, pea fiber or insoluble fraction obtained from peas, etc.) in alkaline solution. As described in the different process variants of the invention detailed below, these polypeptides can be recovered at different stages of the process. Furthermore, the aqueous pea protein composition also comprises proteins precipitated at isoelectric pH.All of the steps involved in providing said aqueous pea protein composition are carried out in a controlled manner, in order to preserve at least a partially native character of the proteins which will be subjected to the heat treatment; as also appears in the Examples section, this is a key criterion in order to obtain the functionalized pea protein of the invention.

[0017] Another subject of the invention relates to the use of the functionalized pea protein according to the invention for the manufacture of food or beverage products, in particular textured proteins by dry or wet extrusion, meat substitutes such as emulsified sausages, charcuterie substitutes, egg substitute compositions or even dessert creams.

[0018] The invention will be described in detail below. Brief description of the drawings

[0019] Other features, details and advantages will become apparent upon reading the detailed description below, and upon analyzing the attached drawings, in which: Fig. 1

[0020] [Fig. 1] shows the residual protein content in the insoluble fraction extracted during aqueous extraction of pea material as a function of extraction pH. Fig. 2

[0021] [Fig. 2] shows the wet extrusion strip Inv. 1 obtained from a mixture comprising the pea protein of Example 1 combined with pea fibers and potato starch obtained with extrusion profile 1. Fig. 3

[0022] [Fig. 3] shows a wet extrusion strip CP.1 obtained from a mixture comprising the pea protein of counterexample 5 obtained with extrusion profile 1. Fig. 4

[0023] [Fig. 4] shows a wet extrusion strip Inv. 2 obtained from a mixture comprising the pea protein of Example 1 obtained with extrusion profile 2. Fig. 5

[0024] [Fig. 5] shows a wet extrusion strip CP.2 obtained from a mixture comprising the pea protein of counterexample 5 obtained with extrusion profile 2. Fig. 6

[0025] [Fig. 6] shows the spoonful texture of the comparative cream obtained from the mixture including the commercial pea protein NUTRALYS® F85 M. Fig. 7

[0026] [Fig. 7] shows the spoonful texture of the cream obtained from the mixture comprising the pea protein of the invention. Fig. 8

[0027] [Fig. 8] shows the spoonful texture of commercial cream. Detailed description of the invention

[0028] The invention relates to a functionalized pea protein which exhibits temperature gelling power and reduced viscosity.

[0029] For the purposes of the invention, a pea protein is an extract from peas comprising mainly proteins and other constituents in lesser proportions, as described later in the description.

[0030] By "functionalized pea protein" is meant a pea protein whose production process makes it possible to provide the gelling power and viscosity of the protein of the invention.

[0031] More precisely, the “gelling power at temperature” is characterized by a gelling power determined according to a TEST A of at least 300 Pa. Gelling power: Test A

[0032] By "gelling power" is meant the functional property consisting of the ability of a protein composition to form a gel or network, increasing viscosity and generating a state of matter intermediate between liquid and solid states. The term "gel strength" can also be used. To quantify this gelling power, it is therefore necessary to generate this network and evaluate its strength. To carry out this quantification, in the present invention, test A 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) Stirring 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.

[0033] Preferably, the imposed stress rheometers are chosen from the DHR 2 (TA, instruments) and MCR 301 (Anton Paar) models, with a concentric cylinder type spindle. They have a Peltier effect temperature control system. To avoid evaporation problems at high temperatures, paraffin oil is added to the samples.

[0034] 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, a gel, a suspension, a paste, etc., in response to an applied force.

[0035] 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 angular velocity œ) and temperature. In particular, this type of rheometer allows access to the complex modulus G*, itself allowing access to the moduli G' or elastic part and G" or viscous part.

[0036] The first three steps consist of resuspending the protein in water, under precise conditions to maximize subsequent measurement.

[0037] The water chosen is preferably osmosis water.

[0038] 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.

[0039] 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 such as beakers and magnetic bars is used. 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 (20°C) using a pH meter and acid-base reagents, as well known in the prior art.

[0040] The fourth step is to introduce the sample into the rheometer, covering it with a thin layer of oil to limit evaporation.

[0041] 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.

[0042] The measurement of the parameter G' is carried out continuously during this scale and is recorded.

[0043] The sixth and final step of test A consists of the exploitation of the recording. Two values ​​are extracted: G'1 = value of G' at the start 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.

[0044] The gelling power is equal to G'2 - G'1.

[0045] Preferably, the gelling power determined according to TEST A ranges from 500 to 2000 Pa, most preferably from 600 to 1800 Pa or from 600 to 1000 Pa or from 1000 to 2000 Pa or from 1100 to 1800 Pa. The gelling power may be greater than or equal to 400 Pa, greater than or equal to 500 Pa, greater than or equal to 600 Pa, greater than or equal to 700 Pa, greater than or equal to 800 Pa, greater than or equal to 900 Pa, greater than or equal to 1000 Pa or greater than or equal to 1100 Pa. The gelling power may be less than or equal to 2000 Pa, less than or equal to 1900 Pa, less than or equal to 1800 Pa, less than or equal to 1700 Pa, less than or equal to 1600 Pa, less than or equal to 1500 Pa, less than or equal to 1400 Pa, less than or equal to 1300 Pa, less than or equal to 1200 Pa, less than or equal to 1100 Pa or less than or equal to 1000 Pa.

[0046] According to the invention, the reduced viscosity of the functionalized pea protein can be expressed by a viscosity of less than 0.65 Pa.s (15% dry matter, 40s-1, 20°C).

[0047] The viscosity can be determined according to the invention according to TEST B. Viscosity: TEST B

[0048] To determine the viscosity profile in water, measurements are carried out on an aqueous solution of pea protein at 15% dry matter (osmosed and azidated water at 200 ppm to prevent any bacteriological risk), in an AR2000 rheometer from TA Instruments, with a concentric cylinder geometry, with a shear rate of 0.6x10' 3 at 600 s -1in 3 minutes (log) and at a temperature of 20°C (3 min of temperature equilibrium before testing). Before measurement, the solution is stirred for at least 10 hours, at 750 rpm and at 20°C. The pH is not adjusted. The viscosity at 40 s-1 is reported as the viscosity according to TEST B.

[0049] The viscosity according to TEST B of the functionalized pea protein may be less than or equal to 0.60 Pa.s, for example less than or equal to 0.55 Pa.s, in particular less than 0.5 Pa.s, advantageously less than 0.4 Pa.s, preferably from 0.05 to 0.4 Pa.s, for example from 0.1 to 0.4 Pa.s or from 0.05 to 0.65 Pa.s, for example from 0.1 to 0.60 Pa.s.

[0050] Preferably, the functionalized pea protein of the invention has a solubility according to a TEST C of less than or equal to 49%, preferably less than or equal to 45%, for example ranging from 20 to 40%.

[0051] Solubility: TEST C

[0052] As for solubility, it is determined according to the TEST C method described below: Measurement of water solubility This measurement is based on diluting the sample in distilled water, centrifuging it and analyzing the supernatant.

[0053] 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 HCl 0.1 N (pH 7). Add water 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 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.

[0054] The soluble solids content, expressed in % by weight, is given by the following formula:

[0055] [Math. 1] (m1 - m2) x (200 + P) x 100 % solubility P1 x P

[0056] 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

[0057] Functionalized pea protein can have a pH ranging from 6.0 to 6.8.

[0058] The functionalized pea protein may have a solubility at pH 7 according to a TEST C of less than or equal to 49%, preferably less than or equal to 45%, preferably 20 to 40%.

[0059] The functionalized pea protein of the invention may have a protein content ranging from 80 to 90%.

[0060] Functionalized pea protein is generally a protein composition comprising, in addition to protein, other minor constituents, such as starch, lipids, fibers, and / or sugars.

[0061] The protein content is the N6.25 content, calculated by the Dumas method. Generally, the total starch content in the pea protein produced according to the method of the invention ranges from 0 to 20%, for example from 0 to 10%, in particular from 0.5 to 5%. This total starch content can be measured using the AOAC 996.11 method. Generally, the total fiber content can range from 0 to 20%, for example from 1 to 18%, in particular from 2 to 10%. This content can be determined by the AOAC Method 2017.16 method. 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 AOAC method 996.06 in acid hydrolysis. The sugar content can range from 0 to 10%, usually 0.5 to 5%. The sugar content can be determined by high-performance liquid chromatography (HPLC).

[0062] All of the above contents are expressed as a function of dry matter.

[0063] According to one embodiment, the functionalized pea protein has a denaturation percentage greater than 75%, or even greater than 90%, most preferably approximately 100%.

[0064] The percentage of denaturation of pea protein is the quantification of denaturation relative to a pea protein isolated by precipitation at isoelectric pH without the addition of heat and subjected to no heat treatment during its manufacture so that it is as native as possible. Thus, to determine the percentage of denaturation, the inventors manufactured a pea protein with an extraction at pH 9 of pea flour in cold water then precipitated at isoelectric pH to pH 5 at room temperature; the precipitated proteins are washed once in cold water to ensure their purity and then dried by lyophilization. Under these preparation conditions, denaturation is considered minimal for a pea protein precipitated at isoelectric pH and this pea protein isolate is considered a standard.

[0065] The enthalpy of denaturation of this standard is used to determine the percentage denaturation of a sample according to the following formula:

[0066] [Math. 2] % denaturation = (AHd standard - AHd sample) / AHd standard x 100

[0067] Depending on the method for determining the enthalpy of denaturation (AHd), the sample AHd and standard AHd may vary. Under the conditions of TEST D described below, the enthalpy of denaturation of this standard measured on this pea protein suspension is 2.4 J / g of suspension, or an enthalpy of the standard protein AHd standard of 12.5 J / g of protein (N6.25) using the N6.25 protein content on its dry matter as well as the dry matter content of the suspension. However, it is quite possible to use another method for determining the enthalpy of denaturation.

[0068] Enthalpy of denaturation: TEST D

[0069] In TEST D, the pea protein is dissolved in water at 20% + / - 2% dry matter and the pH is adjusted to 6.2 if necessary by adding 0.1 N sodium hydroxide or 0.1 N hydrochloric acid. The solution is stirred for 2 hours at 350 rpm at room temperature. The enthalpy of the AH protein is determined by calorimetry (DSC) according to known methods and is expressed per g of protein N 6.25 in the sample. A sample of 10-15 mg of this solution is taken in a hermetically sealed crucible and the enthalpy is determined by calorimetry. The analysis is carried out by heating the suspension of the pea protein extract at 20% dry matter at 10°C / minute from 5 to 120°C. The enthalpy determined for the sample makes it possible to obtain the enthalpy of the protein in J / g of protein using the N6.25 protein content on dry matter as well as the dry matter content.

[0070] DSC calorimeters useful for the measurement can be, for example, the Q20 (TA, instruments), DSC 8000 (Perkin Elmer) and DSC (Mettler) models. The analysis is carried out from 5°C to 120°C at a heating rate of 10°C / minute.

[0071] According to another embodiment, the functionalized pea protein has a residual enthalpy of denaturation of less than 0.5 J / g of N6.25 protein, preferably determined according to a D TEST, or even less than 0.2 J / g of N6.25 protein, or even zero.

[0072] Advantageously, the functionalized pea protein of the invention has a dry matter content greater than 90%, most preferably greater than 94% by weight of dry matter relative to the weight of the functionalized pea protein. The functionalized pea protein may be in the form of a powder having a particle size d50, which may vary widely, for example from 10 to 500 pm, for example from 50 to 500 pm, generally from 50 to 150 pm. The particle size is easily measured by laser diffraction.

[0073] To extract the pea protein of the invention, the inventors developed a controlled extraction process comprising a heat treatment of an aqueous pea protein composition at a determined pH. More specifically, the process for manufacturing the functionalized pea protein of the invention comprises: - a supply of an aqueous pea protein composition comprising pea proteins precipitated at isoelectric pH and pea polypeptides, said polypeptides being insoluble at a pH below 8.5, the pea proteins of the aqueous composition having a percentage of denaturation of less than 65%, - adjustment of the aqueous composition to a pH ranging from 6.0 to 6.8, - a step of heat treatment of the mixture in order to obtain the functionalized pea protein.

[0074] The aqueous pea protein composition takes the form of a suspension comprising mainly water and pea protein. The dry matter of this aqueous composition is easily adjusted by the person skilled in the art depending on the equipment used and can range from 5 to 25%, for example from 10 to 18%.

[0075] According to the invention, the pea proteins from the aqueous composition provided are extracted in such a way as not to significantly denature them. As appears in the remainder of the description, the protein can denature at numerous process steps and, on the basis of the instructions given below, the person skilled in the art will be able to select the conditions of the process of the invention in order to protect the protein during its extraction and thus provide the aqueous pea protein composition useful for the heat treatment step. In general, it will be preferred chaining the different stages of the process in order to preserve, before the heat treatment stage, the undenatured character of the pea proteins included in the aqueous composition. This is all the more important during the stages where there is an input of heat. Preferably, to keep the percentage of denaturation high as desired, cold water, having a temperature ranging from 1 to 25°C, lukewarm water (60°C) or water ranging from 25 to 60°C will be used during the process.

[0076] The pea proteins in the aqueous composition may thus have a denaturation percentage of less than 65% previously described. Advantageously, the pea proteins included in the aqueous composition have a denaturation percentage: - less than 60%, or even less than 55%, or even less than 50% and / or - greater than 10%, or even greater than 20%, or even greater than 30%.

[0077] To determine the percentage of denaturation, the protein formed during the process can be recovered and freeze-dried so as not to denature it, then its enthalpy can be measured and proceeded as explained previously.

[0078] The pea proteins in the aqueous composition comprise pea proteins precipitated at isoelectric pH.

[0079] Isoelectric pH means a pH near which the net electrical charge of the protein in the protein fraction is zero. This pH can be adjusted to a pH between, for example, 4.0 and 5.7, or even 4.8 and 5.2. When a composition comprising pea proteins is placed at this pH, certain proteins, in particular at least some of the globulins, will tend to precipitate. Pea proteins precipitated at isoelectric pH are therefore pea proteins thus precipitated which have been separated by solid-liquid separation. With the exception of the methods described in the TESTS, it is specified that, throughout the document, the pH values ​​are determined at 10% dry matter and at 20°C.

[0080] The pea proteins of the aqueous composition also comprise pea polypeptides, said polypeptides being insoluble at a pH below 8.5. As will be apparent in the remainder of the description, and in particular when discussing the various methods of providing the pea proteins of the aqueous composition, the pea polypeptides may or may not be precipitated at isoelectric pH. In the variants where they are not precipitated at isoelectric pH, the native character of the pea proteins provided before the heat treatment step is increased because the isoelectric precipitation step of the proteins may lead to some denaturation.

[0081] By "pea polypeptides insoluble at a pH below 8.5" is meant a fraction of pea proteins which are extracted into solution (soluble fraction), by placing a pea material in an aqueous composition at a pH above 8.5, for example in a concentration mass of 20% relative to the total weight of the aqueous composition, and which is not extracted at a lower pH.

[0082] When a pea material is put into an aqueous composition to extract proteins, the proteins are solubilized and end up in the aqueous fraction and the non-solubilized proteins remain in the insoluble fraction, along with other insoluble substances such as fiber and starch. However, as shown in Figure 1, during extraction of a pea material, the residual protein content included in the insoluble fraction will decrease when the pH increases: in fact, this is explained by the fact that proteins insoluble at a certain pH are soluble at a higher extraction pH.According to the invention, it is important that the aqueous composition subjected to the heat treatment after pH adjustment has pea proteins solubilized during an alkaline suspension of a pea material in water at a pH greater than 8.5, these proteins therefore being named in the context of the present invention "pea polypeptides insoluble at a pH lower than 8.5". Different embodiments of providing an aqueous composition of pea proteins useful for the method of the invention are described below. They have the common point of comprising an alkaline suspension of a pea material at a pH greater than 8.5. These embodiments should not be considered as limiting the invention.

[0083] The pea material from which the pea polypeptides can be extracted may be any material comprising pea polypeptides that are insoluble at a pH below 8.5. In particular, this pea material may be a pea flour, a pea protein concentrate, a pea fiber or an insoluble fraction obtained from peas.

[0084] According to one embodiment, the pea flour is obtained by dry grinding. Extraction can be carried out by suspending the pea flour thus obtained in water. Suspension of pea flour in water is also understood to mean a suspension obtained by wet grinding of an aqueous suspension of whole peas or cotyledons, advantageously previously dehulled from their outer shell. According to this embodiment, the suspension of pea flour in water can thus be obtained directly. As an example of wet grinding, mention may be made of that described in application WO2019 / 053387.

[0085] Other pea fractions, such as pea fibers, pea concentrates which are obtained by mechanical and dry fractionation of pea flour or other insoluble fractions obtained from peas can be used. In order to extract insoluble pea polypeptides at pH below 8.5, it is obviously preferable that these pea fractions have not undergone prior protein extraction by alkaline suspension at a pH above 8.5.

[0086] Different modes of providing aqueous pea protein composition will now be described.

[0087] According to a first embodiment, the provision of the aqueous pea protein composition comprises: - a suspension of pea flour in water, - separation of the suspension to provide a primary aqueous solution and an insoluble fraction, - suspending at least a portion of the insoluble fraction in water at a pH greater than 8.5 to form an alkaline suspension, - separation of the alkaline suspension to provide a secondary aqueous solution in which the pea polypeptides are solubilized and a residual solid fraction, - mixing the primary aqueous solution and the secondary aqueous solution to form a mixture of primary and secondary aqueous solutions, - setting the mixture of primary and secondary aqueous solutions to isoelectric pH to form the aqueous suspension of pea proteins, comprising the pea proteins precipitated at isoelectric pH and pea polypeptides, - separation of pea proteins from the aqueous pea protein suspension to form the aqueous pea protein composition.

[0088] According to a second embodiment, the provision of the aqueous pea protein composition comprises: - a suspension of pea flour in water, - separation of the suspension to provide a primary aqueous solution and an insoluble fraction, - setting the primary aqueous solution to isoelectric pH to form an aqueous suspension of precipitated pea proteins, - separating the precipitated pea proteins from the aqueous suspension of precipitated pea proteins to form an aqueous composition of precipitated pea proteins, - suspending at least a portion of the insoluble fraction in water at a pH greater than 8.5 to form an alkaline suspension? - separation of the alkaline suspension to provide a secondary aqueous solution which comprises pea polypeptides and a residual solid fraction, - mixing the precipitated pea protein aqueous composition and the secondary aqueous solution to form the pea protein aqueous composition.

[0089] The first and second modes have the advantage of maximizing the recovery of pea polypeptides while limiting the amounts of base required for extraction.

[0090] According to a third embodiment, the provision of the aqueous pea protein composition comprises: -suspending pea flour in alkaline water at a pH above 8.5 to form an alkaline pea flour suspension, - separation of the alkaline suspension of pea flour to provide an aqueous solution comprising pea proteins including pea polypeptides and an insoluble fraction, - setting the aqueous solution to isoelectric pH to form an aqueous suspension of pea proteins comprising the precipitated pea proteins and the pea polypeptides, - separation of the precipitated pea proteins from the suspension to form the aqueous pea protein composition.

[0091] The third mode has the advantage of having fewer unit steps than the other embodiments.

[0092] According to a fourth embodiment, the provision of the aqueous pea protein composition comprises: - suspension of pea flour in water, - separation of the suspension to provide a primary aqueous solution with residues comprising the suspended pea polypeptides and an insoluble fraction, - separating the primary aqueous solution to provide a purified primary aqueous solution and residues comprising the pea polypeptides, - suspending at least a portion of the residues comprising the pea polypeptides in water at a pH greater than 8.5 to form an alkaline suspension, - separation of the alkaline suspension to provide a secondary aqueous solution comprising the pea polypeptides and a residual fraction, - mixing the purified primary aqueous solution and the secondary aqueous solution to form a mixture of aqueous solutions, - setting the mixture of aqueous solutions to isoelectric pH to form an aqueous suspension of pea proteins, - separation of pea proteins from the suspension to form the aqueous pea protein composition.

[0093] According to a fifth embodiment, the provision of the aqueous pea protein composition comprises: - suspension of pea flour in water, - separation of the suspension to provide a primary aqueous solution with residues comprising the suspended pea polypeptides and an insoluble fraction, - separating the primary aqueous solution to provide a purified primary aqueous solution and residues comprising the pea polypeptides, - suspending at least a portion of the residues comprising the pea polypeptides in water at a pH greater than 8.5 to form an alkaline suspension, - separation of the alkaline suspension to provide a secondary aqueous solution comprising the pea polypeptides and a residual fraction, - setting the primary aqueous solution to isoelectric pH to form a protein suspension of rushed peas, - separation of precipitated pea proteins from the suspension of precipitated pea proteins, - mixing the precipitated pea proteins and the secondary aqueous solution to form the aqueous pea protein composition.

[0094] According to a sixth embodiment, the provision of the aqueous pea protein composition comprises: - a suspension of pea flour in water, - separation of the suspension to provide an aqueous solution with residues comprising the suspended pea polypeptides and an insoluble fraction, - suspending at a pH above 8.5 the aqueous solution with the residues comprising the suspended pea polypeptides to form an alkaline suspension, - separation of the alkaline suspension to provide an aqueous solution and a residual solid fraction, - setting the aqueous solution to isoelectric pH to form an aqueous suspension of pea proteins, - separation of pea proteins from the suspension to form the aqueous pea protein composition.

[0095] Generally, the amount of residues comprising the pea polypeptides, relative to the dry matter of the aqueous solution comprising them, ranges from 5 to 20% of the dry matter of said aqueous solution.

[0096] According to a seventh embodiment, the provision of the aqueous pea protein composition comprises: - suspending pea flour in water to form a pea flour suspension, - separation of the pea flour suspension to provide a primary aqueous solution and an insoluble fraction, - separation of the insoluble fraction to form a starch fraction and a fiber fraction, - suspending at least a portion of the fiber fraction in water at a pH greater than 8.5 to form an alkaline suspension, - separation of the alkaline suspension to provide a secondary aqueous solution in which the pea polypeptides are solubilized and a residual solid fraction, - mixing the primary aqueous solution and the secondary aqueous solution to form a mixture of aqueous solutions, - bringing the mixture of aqueous solutions to isoelectric pH to form an aqueous suspension of pea proteins, comprising the pea proteins precipitated at isoelectric pH and the pea polypeptides, - separation of pea proteins from the suspension to form the aqueous pea protein composition.

[0097] According to an eighth embodiment, the provision of the aqueous pea protein composition comprises: - suspending pea flour in water to form a pea flour suspension, - separation of the pea flour suspension to provide a primary aqueous solution and an insoluble fraction, - separation of the insoluble fraction to form a starch fraction and a fiber fraction, - setting the primary aqueous solution to isoelectric pH to form an aqueous suspension of precipitated pea proteins, - separating the precipitated pea proteins from the suspension to form an aqueous composition of precipitated pea proteins, - suspending at least a portion of the fiber fraction in water at a pH greater than 8.5 to form an alkaline suspension, - separation of the alkaline suspension to provide a secondary aqueous solution which comprises pea polypeptides and a residual solid fraction, - mixing the precipitated pea protein aqueous composition and the secondary aqueous solution to form the pea protein aqueous composition.

[0098] The fourth, fifth, sixth, seventh and eighth variants have the advantage of using smaller amounts of base, compared to the other modes, because the alkaline suspension is carried out on a smaller amount of pea material than in the case of flour (third mode) or the entire insoluble fraction resulting from a first separation (first and second modes). The aqueous solution with residues comprising pea polypeptides in suspension is obtained by selecting and adjusting the separation means, that is to say by carrying out the separation so as to keep residues in suspension, these residues comprising the pea polypeptides useful in the invention.

[0099] The separation steps 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 separation systems which use centrifugal force to separate the constituents by using the field of centrifugal forces caused by the movement of the mixture in a fixed device such as hydrocyclones. A person skilled in the art will be able to select the most suitable equipment and conditions of use in order to obtain the different fractions described above.

[0100] Preferably, the aqueous pea protein composition further comprises pea polypeptides insoluble at a pH below 8.7 or below 8.8. Preferably, the aqueous pea protein composition further comprises pea polypeptides insoluble at a pH below 9.0. Preferably, the aqueous pea protein composition further comprises pea polypeptides insoluble at a pH below 9.2. It is readily understood that these insoluble pea polypeptides, similarly to pea polypeptides insoluble at a pH below 8.5, are obtained respectively by an alkaline suspension carried out at a pH above 8.7, a pH above 8.8, a pH above 9.0 and a pH above 9.2.

[0101] To obtain precipitated pea proteins at isoelectric pH, an aqueous solution of solubilized pea protein is generally adjusted to isoelectric pH to form an aqueous suspension of precipitated pea proteins at isoelectric pH, followed by separation of the pea proteins from the suspension. As already mentioned above, the pH can be adjusted between 4.0 and 5.7, or even between 4.8 and 5.2. The pH correction can be carried out by adding acid, organic or inorganic, for example hydrochloric acid, sulfuric acid or citric acid or mixtures thereof. This step e) can be carried out in a tank, stirred or not. It can be more or less long, be almost immediate or last for example from 1 to 240 minutes, generally from 5 to 60 minutes. This addition of base or acid can be carried out online and the acid can be in the form of an aqueous solution.

[0102] During this isoelectric precipitation, it is important to limit the heat input so as to maintain the percentage of denaturation as in the invention. Thus, it is possible to precipitate the protein by thermocoagulation but then this must be done in such a way as to maintain the percentage of denaturation as defined above. It is advantageous to use a pasteurization stage carried out at a temperature below 80°C, advantageously for a short time, which may be less than 30 seconds. Preferably, the pasteurization stage is carried out at a temperature ranging from 45°C to 77°C, advantageously from 60 to 72°C. Preferably, the pasteurization stage has a duration ranging from 1 to 10 seconds. The heat input by direct injection of steam is also preferably limited or even eliminated because it is highly likely to denature the protein.Alternatively, preheating with a heat exchanger, for example a plate heat exchanger followed by heating by direct steam injection, may be used so as to limit the heat input by this direct steam injection. Pasteurization, particularly under the preferred conditions described above, has the advantage of rapidly coagulating the precipitated pea proteins and therefore accelerating the manufacturing process while maintaining the native character of the proteins. Advantageously, in the case of the pasteurization stage, the temperature is rapidly reduced, for example by vacuuming (well known to the person skilled in the art by the term "flash cooling"), the vacuum applied being. determined based on the chosen cooling temperature. This cooling also helps maintain the native character of the pea protein.

[0103] Once supplied, the aqueous pea protein composition is subjected to a pH adjustment ranging from 6.0 to 8.0, preferably from 6.0 to 6.8. This pH adjustment is generally carried out by adding a base, such as sodium hydroxide, potash, lime or a mixture thereof. It is optionally possible to dilute the aqueous composition so that the pH-adjusted aqueous pea protein composition has a dry matter allowing it to be heat-treated in the selected equipment. Preferably, the pH ranges from 6.1 to 6.5. It is preferable that this pH adjustment step between the supply of the aqueous pea protein composition and the heat treatment is carried out as quickly as possible, and has a duration of less than 60 minutes so that the pea protein retains its denaturation enthalpy before the heat treatment step.

[0104] The heat treatment step is then carried out so as to obtain the functionalized pea protein of the invention. A heat treatment is generally characterized by a time / temperature scale. Those skilled in the art will know how to adapt the conditions so as to obtain the functionalized pea protein of the invention.

[0105] The temperature during the step of heat treatment of the mixture can range from 105 to 128°C, preferably from 110 to 125°C, most preferably 120°C. Preferably, the step of heat treatment of the mixture has a duration of 0.1 to 20 seconds, advantageously from 0.1 to 5 seconds. The method of the invention generally comprises, following the heat treatment step, a step of cooling the functionalized 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 can vary according to the temperature of the step preceding it. Generally, the cooling temperature is selected so that the difference between the heat treatment temperature and the cooling temperature ranges from 10 to 80°C, for example from 30 to 70°C.The cooling temperature may range from 30 to 100°C, advantageously is less than 80°C, for example from 60 to 80°C, for example around 70°C. For example, in the advantageous mode where the heat treatment temperature ranges from 105 to 128°C, the cooling temperature may range from 60 to 80°C, for example may be around 70°C.

[0106] According to a variant of the method, it comprises a step of shearing the functionalized pea protein, for example by passing the aqueous dispersion of proteins through a high-pressure pump. As an example of a high-pressure pump, mention may be made of 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. According to another variant, the method comprises a step of homogenization of the functionalized pea protein.

[0107] The shearing or homogenization step can take place before or after the heat treatment and / or pH raising steps. However, if they are carried out, these optional steps are generally carried out at low intensity, without this negatively affecting the functionalities of the pea protein, in particular the gelling power.

[0108] The method according to the invention may also comprise a step of drying the functionalized pea protein. Generally, this drying step is carried out so as to achieve the dry matter content described above. 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 technique may be chosen so that the powder has the particle size d50 described above. The method of the invention may also comprise a step of dry grinding the protein.

[0109] The invention also relates to the use of the functionalized pea protein according to the invention for the manufacture of food or beverage products.

[0110] Generally, the pea 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 based on 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 based on the total dry weight of the food or beverage product. All intermediate amounts (i.e., 2%, 3%, 4%... 77%, 78%, 79% by weight based on the total weight of the food or beverage product) may be used, as may all intermediate ranges based on these amounts. These food and beverage products may be suitable for vegetarian or vegan populations.

[0111] One use of the protein of the invention relates to its use in beverages. 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% by 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 milk alternatives or milk substitutes, including barista-type milks or coffee creamers. The plant-based milk alternatives can be made from the pea protein according to the invention as well as fats, carbohydrates and / or other optional ingredients that are emulsified to form the substitute. Alternatively, the milk alternatives can be made from “plant milks” obtained from plants, such as oat milk, rice milk, soy milk, coconut milk or almond milk. These plant milks can thus be supplemented with the protein of the invention. They can also be other ready-to-drink drinks, 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 drinks such as beers or spirits, smoothies, drink 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 aromatic note or the bitter aftertaste of the protein or to flavor the beverage. One of the advantages of the pea protein of the invention is that its use in place of conventional pea proteins makes it possible to reduce this quantity of flavoring or masking agent, or even to completely remove these constituents from the beverage, while maintaining a very satisfactory taste for the beverage. Beverages may also include hydrocolloids.

[0112] Food products that may be affected include bakery products such as bread products (including, but not limited to, leavened and unleavened breads, sandwich breads, yeast breads and yeast-free breads such as soda breads), breads comprising all types of wheat flour, breads comprising 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, cereal, nut, and / or fruit bars).;

[0113] It can also be gelled desserts such as puddings and custards. A dessert cream means any type of pastry cream, mousseline, diplomat, chiboust, bavaroise or flan. These different creams may be used as fillings. 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 ice cream and all other types of ice cream - and frozen non-dairy desserts such as non-dairy ice cream, sorbet and others).

[0114] Other products conventionally prepared from animal milk may also comprise the pea 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 frais and petits suisses. They may also be cheese substitutes such as spreadable, processed, cooked and uncooked pressed cheeses, soft cheeses, pasta filata, blue-veined cheeses; These include Emmental, string cheese, ricotta, provolone, Parmesan, Munster, mozzarella, Monterey Jack, Manchego, blue cheese, Fontina, feta, Edam, Double Gloucester, Camembert, Cheddar, Brie, Asiago, and Havarti.It can also be other products such as vegetable butters or crème fraîche.

[0115] Other products that may include the pea protein of the invention are also sauces such as salad dressings or mayonnaise or ketchup based sauces or syrups.

[0116] Also, the pea proteins of the invention may be incorporated into confectionery products (including, but not limited to, gummies, 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 may also be sweetened spreads (including, but not limited to, jellies, jams, nut butters such as peanut butter, spreads, and other spreadable products).

[0117] The pea proteins of the invention can also be used as a carrier or in flavor encapsulation.

[0118] Other types of foods and beverages not mentioned herein but which typically comprise one or more proteins may also be contemplated within the scope of the present invention. In particular, animal foods (such as pet foods) are explicitly contemplated.

[0119] Pea protein can also be used, possibly after texturizing, in meat substitutes such as emulsified sausages or hamburgers, cold cut substitutes such as pork, breast, chicken or turkey ham substitutes (also grouped under the name "cold-cut") 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"). 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. Other more specific technologies include shear cell technology, microextrusion, or 3D printing. Pea protein can also be used in the manufacture of binder compositions, which are used in the manufacture of meat or fish substitutes. Binders are often used to assemble textured protein particles to shape these substitutes, which can be done by compression-cooking or by using, for example, PowerHeater™-type shaping equipment marketed by Source Technology.

[0120] Food or beverage products may be used in particular in specialized nutrition, for example for specific populations, e.g., babies or infants, children, adolescents, adults, the elderly, athletes, people suffering from a disease. These may be meal replacement nutritional formulas, complete nutritional drinks, for example for weight management or in clinical nutrition (e.g., tube feeding or enteral nutrition).

[0121] Pea protein can be used as a sole protein source, but can also be used in combination with other supplemental proteins, whether plant or animal. These supplemental proteins can be hydrolyzed or non-hydrolyzed. Typically, these supplemental proteins come in the form of concentrates or isolates.

[0122] The term "plant protein" refers to 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.

[0123] By "legume" is generally meant the family of dicotyledonous plants of the order Fabales. Several legumes are important cultivated plants including soybeans, beans including mung beans, chickpeas, fava beans, peanuts, cultivated lentils, cultivated alfalfa, various clovers, broad beans, carob, licorice and lupin. The additional legume protein may be selected from these legumes or may be a pea protein other than that of the invention. In the present application, the term "cereals" refers to cultivated plants of the grass family producing edible grains, for example wheat, oats, rye, barley, corn, sorghum or rice. Tubers may be carrots, cassava, konjac, potatoes, Jerusalem artichokes, sweet potatoes. Oilseed plants are generally plants that produce seeds from which oil is extracted. Plants Oilseeds 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 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.

[0124] Pea 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.

[0125] The pea protein of the invention is particularly useful for the manufacture of textured proteins by dry or wet extrusion, meat substitutes such as emulsified sausages, cold meat substitutes, egg substitute compositions or even dessert creams.

[0126] 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

[0127] Methods

[0128] Numerous embodiments are exemplified below. As to the methods used, they are described in detail in the description above.

[0129] Series 1 (Examples 1 and 2, counter-examples 1, 2, 3 and 4): Production of pea proteins according to the invention and comparative - Effect of extraction pH and during the production process

[0130] Approximately 900 kg of peas were used. The outer fibers of the peas were first separated from the seeds by crushing (mechanical separation of the outer shell and the pea seed) and dehulling (sorting the outer shells and dehulled pea seeds using compressed air). The crushed seeds are then ground in a high-speed attrition mill to obtain pea flour. The flour is then cooled by compressed air. This flour is then rehydrated with water by a high-shear rotor-stator system to allow rapid and efficient hydration of the flour. This pea suspension is then transferred to a stirred storage tank. Alkalization is optionally carried out using 4% sodium hydroxide at the pH indicated in the Table ("pH extraction"). This suspension of crushed peas fed a decanter centrifuge (Flottweg Z3).The protein fraction was recovered from the so-called overflow (“protein overflow”). The protein fraction was adjusted with hydrochloric acid to pH 5 in a stirred tank and then pasteurized at 70°C for less than 5 minutes. seconds. The protein fraction was then flash cooled. Immediately, the pasteurized protein fraction was passed through a Flottweg Z3 decanter centrifuge. The protein sediment (protein underflow) recovered was diluted in lukewarm water. This sediment was then immediately adjusted to a dry matter of approximately 18% and then rectified to the pH indicated in the Table (pH heat treatment) with 4% sodium hydroxide. The percentage of denaturation of the pea protein is reported in the Table (% denaturation); the percentage of denaturation of the pea proteins in the protein underflow is between approximately 43% and 57%. The pea protein was then heat treated at 120°C for 5 seconds and then flash cooled to approximately 75°C and finally dried in a TGE nozzle atomizer.The pea protein powder recovered and the functionalities analyzed (gelling power, solubility at pH 7, viscosity).

[0131] Example 3: Pea protein according to the invention

[0132] Approximately 900 kg of peas were used. The outer fibers of the peas were first separated from the seeds by crushing (mechanical separation of the outer hull and the pea seed) and dehulling (sorting the outer hulls and dehulled pea seeds using compressed air). The crushed seeds are then ground in a high-speed attrition mill to obtain pea flour. The flour is then cooled by compressed air. This flour is then rehydrated with water by a high-shear rotor-stator system to allow rapid and efficient hydration of the flour. This pea suspension is then transferred to an agitated storage tank. This pea suspension then fed a separation system that uses centrifugal force to separate the constituents using a centrifugal force field caused by the movement of the mixture in a fixed device.This technology allows the separation of a heavy fraction comprising mainly starch and fibers while the light fraction mainly comprises proteins. This light fraction was then passed through a standard decanter centrifuge (Flottweg Z3). A protein fraction from the decanter centrifuge was recovered in the overflow ("protein overflow") while the heavy phase devoid of starch (<2% on dry basis), composed partly of insoluble polypeptides at pH less than 8.5, is also recovered. The protein overflow was adjusted with hydrochloric acid to pH 5 in a stirred tank and then pasteurized at 77°C for less than 5 seconds. The protein fraction was then flash cooled. Immediately, the pasteurized protein fraction was passed through a Flottweg Z3 decanter centrifuge from which the heavy fraction was recovered ("precipitated pea protein underflow").The heavy phase, devoid of starch, is diluted in a 1 to 1 ratio with water before being alkalized to a pH of 9.5. This fraction is again passed through a typical centrifugal decanter system (Flottweg Z3) whose overflow ("polypeptides overflow") is recovered. The heavy fraction "precipitated pea protein underflow" (close to 30% dry matter) is diluted between 18%-. 20% with the overflow "polypeptides". This mixture is then rectified to pH 6.3 with 4% sodium hydroxide. The pea proteins in this mixture have a denaturation percentage of approximately 37%. This rectified mixture was heat treated at 120°C for 5 seconds and then flash cooled to approximately 75°C. This pea protein floc is then dried in a TGE nozzle atomizer. The recovered pea protein powder was then analyzed (gelling power, solubility at pH 7, viscosity).

[0133] Example 4: Pea protein according to the invention

[0134] Approximately 900 kg of peas were used. The outer fibers of the peas were first separated from the seeds by crushing (mechanical separation of the outer hull and the pea seed) and dehulling (sorting the outer hulls and dehulled pea seeds using compressed air). The crushed seeds are then ground in a high-speed attrition mill to obtain pea flour. The flour is then cooled by compressed air. This flour is then rehydrated with water by a high-shear rotor-stator system, to allow rapid and efficient hydration of the flour. This pea suspension is then transferred to an agitated storage tank. This pea suspension then fed a separation system that uses centrifugal force to separate the constituents by utilizing the centrifugal force field caused by the movement of the mixture in a fixed device.This technology allows the separation of a heavy fraction comprising mainly starch and fibers while the light fraction comprising mainly proteins. This fraction is again passed through a typical centrifugal decanter system (Flottweg Z3). A protein fraction was recovered in the overflow ("protein overflow") while a heavy phase devoid of starch (<2% on dry basis), also comprising insoluble polypeptides at pH below 8.5, is also recovered. The heavy phase devoid of starch is then diluted in a 1 to 1 ratio with water to reach a dry matter close to 10% before being alkalized to a pH 9.2. This fraction is again passed through a decanter whose overflow ("polypeptide overflow") is mixed with the protein fraction (protein overflow).This mixture is then acidified with hydrochloric acid to pH 5 before being pasteurized at 70°C for less than 5 seconds and then flash-cooled. Immediately, the pasteurized protein fraction is passed through a decanter centrifuge (Flottweg Z3) from which the heavy fraction called "pea isolate" is recovered and diluted to around 20% with lukewarm water. The pea isolate is then rectified to pH 6.3 with 4% sodium hydroxide. The pea isolate was heat-treated at 120°C for less than 5 seconds and then flash-cooled to around 80°C. This isolate is then dried in a nozzle atomizer. The recovered pea protein powder was then analyzed (gelling power, solubility at pH 7, viscosity).

[0135] Counterexample 5: Pea protein comparison

[0136] This counterexample is almost identical to Example 4 except that it differs in that: - the heavy phase devoid of starch is not used and is eliminated (no recovery of overflow polypeptides), - the protein overflow is acidified with hydrochloric acid to pH 5 before being pasteurized at 77°C for less than 5 seconds and then cooled instantly, - the pea isolate thus recovered is rectified to a pH of 7.7 instead of 6.3.

[0137] The recovered pea protein powder was then analyzed (gelling power, solubility at pH 7, viscosity).

[0138] Example 5: Pea protein according to the invention

[0139] Approximately 900 kg of peas were used. The outer fibers 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). A suspension of peas and water was continuously wet-ground to obtain a suspension of ground peas with approximately 20% dry matter. The suspension was then transferred to a stirred storage tank before being adjusted to pH 9.2 by adding 4% sodium hydroxide. This suspension of ground peas fed a decanter centrifuge (Flottweg Z3). The protein fraction was recovered in the overflow. The protein fraction was adjusted with hydrochloric acid to pH 5 in a stirred tank and then pasteurized at 70°C for less than 5 seconds and immediately the pasteurized protein fraction was passed through a Flottweg Z3 decanter centrifuge.The recovered protein sediment (underflow) was diluted in warm water. This sediment was then adjusted to a dry matter content of approximately 18% and then rectified to pH 6.3 with 4% sodium hydroxide and was heat-treated at 120°C for 5 seconds and then cooled by rapid cooling ("flash cooling") to approximately 75°C. The protein was atomized in a TGE nozzle atomizer. The recovered pea protein powder was then analyzed (gelling power, solubility at pH 7, viscosity).

[0140] Example 6: Pea protein according to the invention

[0141] Example 6 is reproduced identically to Example 3 except that the pH during heat treatment is set to 6.65.

[0142] In addition to the proteins of the examples and counterexamples, the gelling power and viscosity of the commercial pea protein isolates PISANE F9, C9 and M9, marketed by the company COSUCRA, were reported.

[0143] [Table 1]

[0144] As appears above from the properties of the different examples, counter-examples and commercial products, the invention has made it possible to obtain pea proteins having a unique combination of properties, a priori incompatible: in fact, using the process mastered during the different manufacturing stages, the inventors have succeeded in obtaining pea proteins combining gelling power at temperature as well as low viscosity. However, the prior art teaches that obtaining proteins with increased gelling power at temperature is accompanied by a strong increase in viscosity at room temperature. All pea proteins according to the invention have zero residual denaturation enthalpies.

[0145] These new proteins will now be evaluated below in different applications.

[0146] Evaluation of the pea protein according to the invention in wet extrusion

[0147] A mixture of powders is made according to the following recipes described in the Table below, expressed in mass of the ingredients:

[0148] [Table 2]

[0149] The proteins tested are the pea protein from example 1 as well as the pea protein from counterexample 5.

[0150] This mixture is introduced by gravity into a LEISTRITZ ZSE 27MAXX extruder from the LEISTRITZ company.

[0151] The mixture is introduced at a regulated flow rate of approximately 13.3 kg / h. A quantity of approximately 15.3 kg / h of water is also introduced. The humidity in the extruder is approximately 56%.

[0152] The wet extrusion tests are carried out on this extruder equipped with a thermoregulated die, model FDK750 from Coperion, comprising two modules of length 80 cm and passage section 50 mm x 15 mm. The extrusion screw is rotated at a speed equal to 350 rpm and sends the mixture into the die. 2 temperature profiles were used according to the test.

[0153] Extruder temperature profile 1, equipped with 15 heatable barrels, is detailed below:

[0154] [Table 3]

[0155] Extruder temperature profile 2 is detailed below:

[0156] [Table 4]

[0157] The textured protein thus produced is cut at the outlet of the die into strips approximately 10 cm long (width 5 cm and thickness 1.5 cm).

[0158] For the 4 tests, the extrusion parameters are reported below:

[0159] [Table 5]

[0160] Observation of band fibration

[0161] To observe the fiberization of the strip, the strip is cut in half lengthwise and pulled between the two pieces of the strip so as to tear it and see the presence or absence of fibers. The torn strips are observed in Figures 2 to 5.

[0162] The CP 1 test was not conclusive: the extrusion was not carried out in a stable manner: the product was torn at the extruder outlet and a significant separation between the center and the shell was observed, as shown in Figure 3. On the contrary, the test of Example 1 shows good fibration (Figure 2).

[0163] At a lower temperature, the CP 2 test shows that it is possible to obtain correct fibration of the comparative mixture at a lower temperature, but that this remains limited (Figure 5). This fibration is higher, with the same temperature profile, using the mixture using the protein of the invention (Figure 4). It is also observed that the pressure increases in the extruder for the 2 tests compared to the hotter temperature profile but that this is not too high to conduct the tests.

[0164] Thus, by using the pea protein of the invention and unlike the comparative protein, it is possible to use varied temperature profiles. In particular, it was possible to use a higher temperature profile (profile 1 includes areas with a maximum of set temperature at 150°C) and to provide stable extrusion bands whereas this was not possible with the comparative protein. Furthermore, the use of the pea protein of the invention made it possible to observe better fibration of the extrusion bands, in particular by using this higher temperature profile.

[0165] Evaluation of the pea protein according to the invention for the manufacture of dessert creams

[0166] The protein of the invention evaluated is the protein of example 4. The comparative protein used is a commercial pea protein (NUTRALYS® F85M marketed by the Applicant).

[0167] The proteins of the invention and comparative were evaluated in the following dessert cream recipe. In the rest of the text, the comparative cream is obtained using the comparative pea protein and the "invention" cream is obtained using the pea protein according to the invention.

[0168] [Table 6]

[0169] This recipe is carrageenan-free.

[0170] Experimental protocol: 1 / Hydrate the protein powders in water at 55°C for 30 minutes with a Silverson mixer at 2500 rpm 2 / Add the rest of the powders and mix at 4000 rpm for 5 minutes 3 / Add the coconut oil and mix at 6000 rpm for 5 minutes 4 / Homogenize at 100 bar at 57°C 5 / Sterilize at 130°C for 30 seconds then cool to 75°C 6 / Leave to cool to room temperature (20°C) 7 / Store at 4°C.

[0171] It was possible to observe a development of the consistency of the dessert cream following its sterilization and its much greater cooling with the protein of the invention. However, despite this phenomenon, no implementation problems during mixing and homogenization were observed.

[0172] 9 days after the creams were made, a sensory evaluation was carried out.

[0173] As a reference point, the properties were also compared with those of a commercial product comprising dairy products without pea protein and carrageenans (Danette® vanilla flavor from the Danone company).

[0174] The conclusions are reported below:

[0175] [Table 7]

[0176] Thus, it was possible with the protein of the invention to formulate a dessert cream almost as thick and firm as commercial dessert cream, while the latter includes milk proteins known to be generally more gelling than pea proteins, as well as carrageenans which are thickeners known to provide significant texture when added to the cream. This makes it possible to have creams with a texture in the mouth and on the spoon very close to the commercial product.

[0177] Moreover, both the inventive and comparative dessert creams have a satisfactory taste.

Claims

Claims

1. Functionalized pea protein having a gelling power according to TEST A of at least 400 Pa, preferably ranging from 500 to 2000 Pa, most preferably ranging from 600 to 1800 Pa and a determined viscosity of less than 0.65 Pa.s, the viscosity being measured at 15% by weight of dry matter, at a shear rate of 40s-1 and a temperature of 20°C.

2. Functionalized pea protein according to claim 1 characterized in that it has a gelling power according to TEST A ranging from 600 to 1000 or from 1000 to 2000 Pa or from 1100 to 1800 Pa.

3. Functionalized pea protein according to one of claims 1 to 2, characterized in that it has a pH ranging from 6.0 to 6.

8.

4. Functionalized pea protein according to one of claims 1 to 3 characterized in that it has a viscosity of less than 0.5 Pa.s (15% dry matter, 40s-1, 20°C), advantageously less than 0.4 Pa.s, preferably from 0.05 to 0.4 Pa.s, preferably from 0.1 to 0.4 Pa.s.

5. Functionalized pea protein according to one of claims 1 to 4 characterized in that its solubility at pH 7 according to a TEST C is less than or equal to 49%, preferably less than or equal to 45%, preferably 20 to 40%.

6. Functionalized pea protein according to one of claims 1 to 5 characterized in that its protein content ranges from 80 to 90%.

7. Functionalized pea protein according to one of claims 1 to 6, characterized in that it has a residual denaturation enthalpy of less than 0.5 J / g of N6.25 protein, or even less than 0.2 J / g of N6.25 protein, or even zero.

8. Use of the functionalized pea protein according to one of claims 1 to 7 for the manufacture of food or beverage products, in particular textured proteins by dry or wet extrusion, meat substitutes such as emulsified sausages, charcuterie substitutes, egg substitute compositions or even dessert creams.

9. Process for manufacturing a functionalized pea protein according to one of claims 1 to 7, characterized in that it comprises: - a supply of an aqueous pea protein composition comprising pea proteins precipitated at isoelectric pH and pea polypeptides, said polypeptides being insoluble at a pH below 8.5, the pea proteins of the aqueous composition having a percentage of denaturation of less than 65%, - adjustment of the aqueous composition to a pH ranging from 6.0 to 6.8, - heat treatment of the mixture to obtain the functionalized pea protein.

10. Manufacturing method according to claim 9 characterized in that the provision of the aqueous pea protein composition comprises: - suspension of pea flour in water, - separation of the suspension to provide a primary aqueous solution with residues comprising suspended pea polypeptides and an insoluble fraction, - separating the primary aqueous solution to provide a purified primary aqueous solution and residues comprising pea polypeptides, - suspending at least a portion of the residues comprising pea polypeptides in water at a pH greater than 8.5 to form an alkaline suspension, - separation of the alkaline suspension to provide a secondary aqueous solution comprising the pea polypeptides and a residual solid fraction, - setting the primary aqueous solution to isoelectric pH to form a suspension of the precipitated pea proteins, - separation of precipitated pea proteins from the suspension of precipitated pea proteins, - mixing the precipitated pea proteins and the secondary aqueous solution to form the aqueous pea protein composition.

11. Method according to claim 10 characterized in that the isoelectric precipitation step comprises a pasteurization stage carried out at a temperature below 80°C, advantageously from 45°C to 77°C, advantageously for a time below 30 seconds, preferably for a duration of 1 to 10 seconds.

12. Method according to one of claims 10 or 11, characterized in that the alkaline suspension is carried out at a pH greater than 8.7, or greater than 8.8, or even greater than 9.0 or even a pH greater than 9.

2.

13. Method according to one of claims 9 to 12, characterized in that the temperature during the step of heat treatment of the mixture ranges from 105 to 128°C, preferably from 110 to 125°C, most preferably 120°C, this step advantageously having a duration ranging from 0.1 to 20 seconds, advantageously from 0.1 to 5 seconds.

14. Method according to one of claims 9 to 13 characterized in that the pea proteins included in the aqueous composition have a percentage of denaturation: - less than 60%, or even less than 55%, or even less than 50% and / or - greater than 10%, or even greater than 20%, or even greater than 30%.

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