Functional fava bean protein

Abstract

Process for manufacturing a denatured faba bean protein extract, a denatured faba bean protein extract, a denatured faba bean protein composition, a textured faba bean protein composition, and on the use of said extract and said compositions.

Description

Title of the invention: Functional faba bean protein Field of invention

[0001] The invention relates to novel fava bean proteins with improved functional properties, which may be a water solubility profile, an emulsifying capacity and / or a gelling power. The invention also relates to a method for extracting fava bean proteins. Another subject also relates to the use of fava bean proteins in food or beverage products. Prior art

[0002] Field beans, or fava beans (according to the old spelling), are annual plants of the species Vicia faba. They are legumes of the family Fabaceae, subfamily Faboideae, tribe Fabeae.

[0003] This is the same species as the broad bean, a plant used since ancient times for human consumption. The word broad bean then refers to both the seed and the plant.

[0004] Several production processes are known from the prior art which make it possible, starting from field bean seeds, to produce protein compositions.

[0005] A first family of this type of composition relates to protein concentrates, generally obtained in a so-called "dry" process in which the beans are ground to obtain a flour, and then this flour is enriched with protein by mechanical separation. This type of bean protein concentrate has a protein content which does not exceed 70% and which rarely exceeds 60%. In this type of process, no heat treatment step is generally carried out and the proteins in these compositions are generally native and not denatured.

[0006] A second family of this type of composition concerns protein isolates, obtained by a wet process. The conventional wet process begins with grinding the fava beans to obtain a flour. This is then diluted in water to undergo an extraction aimed at solubilizing the fava bean proteins. The solution then undergoes a liquid / solid separation in order to obtain, on the one hand, a crude protein solution and, on the other hand, a solid fraction enriched in starch and fibers. The fava bean proteins are extracted via precipitation at isoelectric pH of the proteins, then they are generally separated from the aqueous solution and dried.

[0007] However, to commercially exploit these proteins obtained by a wet process, manufacturers carry out one or more heat treatment steps in order to pasteurize these proteins as well as a drying step by adding heat, generally by spray drying. These steps are mandatory to be able to control their microbiology and thus allow them to be consumed safely and preserved for several months. However, by the application of sufficient heat to achieve this, these steps inevitably lead to the denaturation of proteins. However, by proceeding with their denaturation, it has been observed in the case of faba bean proteins that the properties obtained are not always satisfactory.

[0008] This is how Vogelsang-O'Dwyer et al. describe in a 2020 article (Vogelsang-O'Dwyer et al. Comparison of Faba Bean Protein Ingredients Produced Using Dry Fractionation and Isoelectric Precipitation: Techno-Functional, Nutritional and Environmental Performance, Foods 2020, 9, 322) the manufacture of a faba bean protein concentrate as well as the manufacture of a faba bean protein isolate. As for the protein concentrate manufactured by mechanical separation, it has a high solubility at pH 7, close to 90%. On the contrary, the faba bean protein isolate has a solubility of only 30%. The authors explain this difference by the denaturation of proteins during the protein isolation and drying steps, which are carried out using heat for the isolate, while the protein in the concentrate is not denatured.

[0009] Furthermore, Johnston et al. describe in a 2015 article (Johnston et al. The physicochemical properties of legume protein isolates and their ability to stabilize oil-in-water emulsions with and without genipin, J Food Sci Technol. 2015 Jul; 52(7): 4135-4145) the manufacture of different protein isolates, including a faba bean protein isolate. This isolate has a solubility of approximately 85%. However, the protein in this isolate is not denatured because the extraction is carried out at room temperature and the drying is carried out by lyophilization and not by heat application. And above all, this type of undenatured faba bean protein, dried by cryogenization, is not easily exploitable industrially: in fact, this process does not include any step allowing the microbial load of the protein to be controlled in order to consume it directly in food products in complete safety.However, such a protein whose microbial control has not been carried out in its process is not industrially and commercially acceptable. Another process for manufacturing undenatured or weakly denatured legume protein, which may be from faba bean, has also been described in document WO2022 / 136627. However, although a heat treatment step can be applied, it is essential that the protein remains undenatured and the heat treatment must remain low intensity; this document therefore does not solve the problem of good control of the microbial load. This is also the case for application WO 2023 / 073238 which describes a faba bean protein comprising albumins and globulins. This composition is obtained by a process using ultrafiltration, not including isoelectric precipitation and optionally a low temperature heat treatment, so that the protein remains native.

[0010] The Applicant has already been interested in providing new protein compositions with improved color which were the subject of applications WO2020 / 193641 and WO2020 / 193668. However, these documents describe denatured faba bean proteins whose properties can be further improved.

[0011] However, there are many applications for which the use of such proteins with improved properties may be of interest. For example, in the field of ready-to-drink beverages and for example for the manufacture of vegetable milk substitutes, obtaining a soluble protein makes it easier to prepare a beverage with a greater mouthfeel. The beverage is more pleasant to taste for the end consumer. Also, in the field of powdered beverages (or powder mixes), it is imperative to have good solubility to avoid the powdery texture in the mouth which is unpleasant for the consumer. The majority of these beverages are neutral or even very slightly acidic and have a pH ranging from approximately 6 to approximately 7.However, it has been observed that during the denaturation of faba bean proteins, their solubility profile is modified and that the solubility decreases whatever the pH, and in particular the solubility at pH ranging from 6 to 7. This can be a problem because, in addition to beverages, many food products have such a slightly acidic pH close to 6. As shown in the Examples section, the currently available commercial faba bean proteins have relatively low solubility in this pH range, which can limit their use. For example, the texture of the protein may be considered unpleasant, rough in the mouth, because it lacks solubility at the pH of use.It is therefore desirable to provide new fava bean proteins that can exhibit a different and improved solubility profile compared to known fava bean proteins, in order to be able to optimize the use of fava bean proteins, particularly in food applications.

[0012] As another functional property, the emulsifying capacity of available faba bean proteins can also be quite limited, lower for example than certain pea proteins. However, many food products (dairy substitutes, sauces, meat substitutes, etc.) combine lipids and water and obtaining proteins with this superior emulsifying capacity would make it possible to obtain improved food products. This is also the case for ice creams and ice creams, in which the quantities of lipids are significant, for which such proteins are desired.

[0013] It may also be of interest to provide proteins combining these different properties, including solubility and emulsifying capacity mentioned above. Another useful functional property that may be mentioned is gelling power, which is useful for the manufacture of food products with a greater mouthfeel. However, gelling power is generally associated with relatively low solubility, particularly at neutral pH. It may be of interest to provide new faba bean proteins combining good gelling power with high solubility, in order to obtain products with different textures.

[0014] It is therefore of interest for the commercial development of fava bean proteins to find a simple and effective process, allowing access to new fava bean proteins having functional properties (or a combination of these properties) such as solubility, emulsifying capacity and / or improved gel strength compared to the fava bean proteins of the prior art.

[0015] It is to the applicant's credit to have found such a method and such a protein composition. This invention will be described in the following section. Summary of the invention

[0016] In a first subject, the invention relates to a method for manufacturing a denatured faba bean protein extract comprising the following steps: a) preparation of an aqueous suspension of crushed field beans; b) extraction of a protein fraction by solid-liquid separation of the aqueous suspension; c) adjustment to a pH between 4.0 and 5.7 of said protein fraction; d) heat treatment of the protein fraction at a temperature ranging from 50 to 80°C for a time ranging from 1 to 30 seconds to form a suspension of precipitated proteins; e) solid-liquid separation of the suspension of precipitated proteins to recover a field bean protein extract having a protein content by weight relative to its dry matter greater than 70%; f) pH adjustment between 6.3 and 8; g) heat treatment of the fava bean protein extract at pH adjusted to a temperature ranging from 85 to 160°C for a period of time sufficient to obtain denaturation of the fava bean protein; h) possibly drying.

[0017] In certain embodiments, the method comprises, following the heat treatment step g), a step of vacuum cooling the suspension of precipitated proteins.

[0018] In certain embodiments, the heat treatment step g) is carried out at 100 to 150°C for 0.1 to 60 seconds, preferably 0.1 to 10 seconds.

[0019] In certain embodiments, the method comprises, after step g), a step of shearing the denatured protein extract, for example by passing it through a high pressure or homogenization pump.

[0020] In certain embodiments, the method is characterized in that a starch-rich fraction and / or a fiber-rich fraction is also recovered from the aqueous suspension of ground fava beans.

[0021] In some embodiments, the pH is adjusted between 6.5 and 7.5 during step f).

[0022] In a second subject, the invention relates to a denatured field bean protein extract capable of being obtained by the method of the first subject of the invention.

[0023] In a third subject, the invention relates to a denatured faba bean protein composition, comprising at least 60% by weight of proteins on a dry matter basis, said composition having at least one of the following properties: - solubility at pH 6 greater than or equal to 50%, - solubility at pH 7 greater than or equal to 75%, - emulsion capacity greater than 400 mL / g.

[0024] In certain embodiments, the composition has a solubility at pH 6 greater than or equal to 70% or a solubility at pH 7 greater than or equal to 80%.

[0025] In certain embodiments, the composition has an emulsion capacity greater than 500 mL / g, preferably greater than 600 mL / g.

[0026] In certain embodiments, the composition has a gelling power greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa.

[0027] In certain embodiments, the composition has a faba bean protein content greater than 80%, preferably greater than 90%, % expressed by weight relative to the dry matter of the composition.

[0028] In certain embodiments, the composition has a dry matter greater than 90%, preferably greater than 94%, % expressed by weight relative to the total weight of the composition.

[0029] In some embodiments, the composition has a denaturation enthalpy of less than 0.5 J / g.

[0030] In a fourth subject, the invention relates to a textured fava bean protein composition obtained by texturizing the fava bean protein extract of the second subject of the invention, or the fava bean protein composition of the third subject of the invention.

[0031] In a fifth subject, the invention relates to the use of the fava bean protein extract of the second subject of the invention, of the denatured fava bean protein composition of the third subject of the invention, or of the textured fava bean protein composition of the fourth subject of the invention, for the manufacture of food or beverage products.

[0032] As will be seen hereinafter, the novel faba bean protein extract has many advantageous uses in beverages and food products, in particular for the manufacture of dairy substitute products (such as yogurts, animal milks or even ice creams), meat substitute products (such as sausages) and in particular use in wet extrusion. Description of the drawings Fig.l

[0033] [Fig.l] shows a wet extrusion strip obtained from a mixture Inv. 1 comprising the faba bean protein of Example 1 combined with pea fibers and potato starch. Fig. 2

[0034] [Fig.2] shows a wet extrusion strip obtained from a CP.l mixture including commercial pea protein. Fig. 3

[0035] [Fig.3] shows a wet extrusion strip obtained from a mixture Inv.2 comprising only the faba bean protein of Example 1. Fig. 4

[0036] [Fig.4] shows a wet extrusion strip obtained from a mixture Inv.3 la faba bean protein of Example 1 in combination with comparative pea protein. Fig. 5

[0037] [Fig.5] shows the results of the tearing test of a wet extrusion strip obtained from a mixture Inv.l comprising the faba bean protein of Example 1 combined with pea fibers and potato starch. Fig. 6

[0038] [Fig.6] shows the results of the tearing test of a wet extrusion strip obtained from a CP.l mixture comprising a commercial pea protein. Fig. 7

[0039] [Fig.7] shows the results of the tearing test of a wet extrusion strip obtained from a mixture Inv.2 comprising only the faba bean protein of Example 1. Fig. 8

[0040] [Fig.8] shows the results of the tearing test of a wet extrusion strip obtained using a mixture Inv.3 comprising the faba bean protein of Example 1 in combination with commercial pea protein. Detailed description of the invention

[0041] A first subject of the invention relates to a process for preparing a denatured faba bean protein extract. This process is described in detail later in the description.

[0042] A second subject of the invention relates to a denatured fava bean protein extract obtainable by the method of the first subject of the invention

[0043] This extract may have different functional properties, in particular in terms of solubility, emulsifying capacity and / or gel strength, which are preferably improved compared to the fava bean proteins of the prior art.

[0044] By "denatured fava bean protein extract", "fava bean protein extract", "composition based on a fava bean protein extract", "fava bean protein composition", or more simply "fava bean protein", is meant a protein composition extracted from fava bean seeds, which can be produced according to the method of the first subject of the invention. Those skilled in the art will understand that in the following these terms can be used interchangeably. The meaning to be given to the term "denatured", relating to fava bean proteins is described later in the description.

[0045] Preferably, the faba bean protein extract according to the invention has a protein content by weight greater than 70% expressed as a percentage of protein on dry matter, preferably greater than 80% by weight, even more preferably greater than 90% by weight. This weight content may range, for example, from 80 to 99%, in particular from 85 to 98%, for example from 90 to 97%. The protein content is the N6.25 content, calculated by the Dumas method.

[0046] According to the invention, unless explicitly stated otherwise, the contents are contents by weight expressed relative to the total dry matter of the extract. The dry matter can be determined using known moisture analyzers, such as an infrared or halogen radiation moisture analyzer.

[0047] The faba bean protein extract according to the invention may comprise, in addition to the faba bean proteins, other minor constituents, such as starch, lipids, fibers, and / or sugars. Generally, the total lipid content ranges from 0 to 15%, in particular from 1 to 10%, for example from 4 to 8%. The total lipid content may be determined by conventional methods, such as the method described in the examples section. The sugar content may range from 0 to 10%, generally from 0.5 to 5%. The sugar content may be determined by high performance liquid chromatography (HPLC). Generally, the total starch content in the faba bean protein extract 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 (2005) method. Generally, the total fiber content can range from 0 to 20%, for example, from 1 to 18%, especially from 2 to 10%.This content can be determined by the AOAC 2017.16 (2017) method.

[0048] According to a variant of the invention, the field bean protein extract of the invention has a particular solubility profile.

[0049] Solubility at pH 6

[0050] According to a preferred embodiment, the faba bean protein extract of the invention has a solubility at pH 6, preferably measured according to test A, ranging from 50% to 100%. The solubility may have all intermediate amounts (i.e. 50%, 51%, 52%... 97%, 98%, 99%) and those skilled in the art will know, on the basis of the process indications indicated in the description and in particular in the Examples section, how to modify the process parameters within the ranges indicated in order to achieve the desired solubility. Advantageously, the solubility at pH 6, preferably measured according to test A, is greater than or equal to 60%, greater than or equal to 65%, or even greater than or equal to 70% or even greater than or equal to 75%. The solubility at pH 6 of the faba bean protein extract of the invention, preferably measured according to test A, may be at most 100%.According to the invention, the solubility of the field bean protein extract at pH 6 may be less than or equal to 90%, for example less than or equal to 85%.

[0051] Solubility at pH 7

[0052] According to one embodiment, the fava bean protein extract may have a solubility at pH 7, preferably measured according to test A, ranging from 75 to 100%. The solubility may have all intermediate amounts (i.e. 76%, 77%, 78%... 97%, 98%, 99%) and those skilled in the art will know, on the basis of the process indications indicated in the description and in particular in the Examples section, how to modify the process parameters within the ranges indicated in order to achieve the desired solubility. Preferably, the fava bean protein extract has a solubility at pH 7, preferably measured according to test A, greater than or equal to 80%, or even greater than 85%. The solubility at pH 7 of the faba bean protein extract of the invention, preferably measured according to test A, may be at most 100%. According to the invention, the solubility of the faba bean protein extract at pH 7 may be less than or equal to 98%, for example less than or equal to 96%.

[0053] According to the solubility profile of the protein extract of the invention, the solubility at pH 6 is generally lower than the solubility at pH 7. The faba bean protein extract of the invention may combine the above-mentioned parameters of solubility at pH 6 and solubility at pH 7.

[0054] Solubility: Test A

[0055] In the present invention, the solubility at pH 6 or pH 7 is determined according to the method of Test A described below.

[0056] This measurement is based on diluting the sample in distilled water, centrifuging it and analyzing the supernatant.

[0057] 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 a sample, possibly adjusted to 95% dry matter, - Adjust or not the pH to the desired value with NaOH or HCl 0.1 N (pH 6 or pH 7), - Complete the water content to 200 g, - Mix for 30 minutes at 1000 rpm and centrifuge for 15 minutes at 3000 g, - Collect 25 g of the supernatant, - Introduce into a previously dried and tared crystallizer, - Place in an oven at 60°C until the visible water has evaporated, - Then place in the oven at 105°C for 1 hour or until constant weight, - Then place in a desiccator (with dehydrating agent) to cool to room temperature and weigh, - The soluble solids content, expressed in % by weight, is given by the following formula:

[0058] [Math.l] % solubility = PlxP Or: - P = weight, in g, of the sample = 5 g - ml = weight, in g, of the crystallizer after drying - m2 = weight, in g, of the empty crystallizer - PI = weight, in g, of the collected sample = 25 g

[0059] Denatured protein

[0060] According to the invention, the term “denatured” protein means a protein which has significantly lost its native character.

[0061] Advantageously, the denatured faba bean protein of the invention has a denaturation enthalpy of less than 1 J / g, or even less than 0.5 J / g or even does not have an enthalpy peak. This enthalpy is expressed in J / g of protein N 6.25.

[0062] The denaturation enthalpy of the denatured faba bean protein (denoted DH) can be determined by calorimetry, in particular by differential scanning calorimetry (DSC) according to methods known to those skilled in the art. For example, the enthalpy can be determined by DSC calorimetry by heating at 10°C / minute from 5 to 120°C a suspension of the faba bean protein extract at 20% dry matter.

[0063] According to one method, the protein extract is dissolved in water at 20% + / - 2% dry matter, the pH being optionally adjusted to 7 by adding 0.1N NaOH or 0.1N HCl. The solution is stirred for 2 hours at 350 rpm at room temperature. A sample of 10-15 mg of this solution is taken in a hermetic crucible then sealed and the enthalpy is determined by calorimetry and from this weight of solution is deduced, using the mass concentration of the solution and the protein content N6.25 in the protein extract.

[0064] DSC calorimeters 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. A person skilled in the art can easily quantify the enthalpy from the thermogram by integrating the endothermic peak, generally using the calorimeter control software. The start of denaturation is expressed by the Tonset (°C) and the end of Tendset (°C). The critical denaturation temperature Tpeak (°C) is determined at the top of the peak.

[0065] Emulsion capacity

[0066] Another advantage of the faba bean protein extract of the invention is that it can have an emulsion capacity preferably determined according to a Test B greater than 400 mL / g. Advantageously, the emulsion capacity is greater than 500 mL / g, preferably greater than 600 mL / g. It is generally less than 1000 mL / g, often less than 900 mL / g, for example less than 800 mL / g. This emulsion capacity is particularly advantageous when it is desired to use the protein of the invention to formulate products comprising fats and in particular emulsions.

[0067] Emulsion capacity: Test B

[0068] In the present invention, the emulsifying capacity is preferably determined by Test B described below.

[0069] Test B determines the ability of a product to emulsify a maximum quantity of oil.

[0070] Operating mode: 0.2 g of sample, possibly adjusted to 95% dry matter, is dispersed for 30 seconds in a volume of water of 60 mL using an Ultraturax set at 9500 rpm, - Gradually incorporate while stirring (9500 rpm) an equivalent volume of corn oil (60 mL), emulsify for 5 min in total, - Centrifugation of a fraction of the mixture at 4000 rpm for 5 minutes, - If emulsion: increase the volumes of water and oil by 10 mL each, respecting the proportion of 50% / 50%, then re-emulsify and centrifuge as above, - If phase shift: reduce the volumes of water and oil by 10 mL each, respecting the proportion of 50% / 50%, then re-emulsify and centrifuge as above, - The result corresponds to the last tube in which the emulsion is observed not to separate after centrifugation.

[0071] Viscosity

[0072] Generally, the field bean protein extract of the invention has, whatever the variant, a viscosity less than or equal to 1 Pa.s, advantageously less than or equal to 0.6 Pa.s, for example ranging from 0.05 to 0.6 Pa.s, or from 0.1 to 0.55 Pa.s, for example from 0.2 to 0.5 Pa.s. This viscosity is an advantage when one wishes to use the faba bean protein extract of the invention to formulate products with a soft texture.

[0073] The viscosity is determined using a plane-plane rheometer, with a 15% aqueous solution of dry matter of faba bean protein extract at a temperature of 20°C, at a shear rate of 40 s1.

[0074] Viscosity: Test C

[0075] For example, viscosity can be determined according to Test C described below.

[0076] For the determination of the viscosity profile in water, the measurements can be carried out carried out: - solubilization of the sample to be tested so as to obtain an aqueous solution at 15% dry matter (preferably prepared with osmosis water and azidated to 200 ppm to prevent any bacteriological risk), - in AR2000 rheometer from TA Instruments, - with concentric cylinder geometry, - with a shear rate of 0.6 103 at 600 s 1 in 3 minutes (log) and - at a temperature of 20°C (3 min of temperature equilibrium before testing).

[0077] Before measurement, the solution is stirred for at least 10 hours, at 750 rpm and at 20°C. The pH is not adjusted.

[0078] The result of Test C corresponds to the viscosity recorded for a shear rate of 40 s A

[0079] Gelling power

[0080] The faba bean protein extract of the invention may have a gelling power greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa. Those skilled in the art may vary the gelling power by varying the process conditions as described later in the description and in the examples section. By "gelling power" is meant the functional property consisting of the capacity of a protein composition to form a gel or a network, increasing the viscosity and generating a state of matter intermediate between the liquid and solid states. 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.

[0081] Gelling power: Test D

[0082] To carry out this quantification, in the present invention, the D test is preferably used, the description of which is as follows: 1) Solubilization at 60°C + / - 2°C of the sample to be tested in water so as to obtain a solution titrating 15% + / - 2% in 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.

[0083] Preferably, the imposed stress rheometers are chosen from the DHR 2 (TA, instruments) and MCR 301 (Anton Paar) models, with a concentric cylinder type mobile. They have a Peltier effect temperature regulation system. In order to avoid evaporation problems at high temperature, paraffin oil is added to the samples.

[0084] A “rheometer” within the meaning of the invention is a laboratory device capable of making measurements relating to the rheology of a fluid or a gel. It applies a force to the sample. Generally of small characteristic dimension (very low mechanical inertia of the rotor), it allows the mechanical properties of a liquid, a gel, a suspension, a paste, etc., to be fundamentally studied in response to an applied force.

[0085] The so-called "imposed constraint" models make it possible, by applying a sinusoidal stress (oscillation mode), to determine the intrinsic viscoelastic quantities of the material, which depend in particular on time (or the angular speed co) and the temperature. In particular, this type of rheometer makes it possible to access the complex module G*, itself making it possible to have access to the modules G' or elastic part and G" or viscous part.

[0086] The first three steps consist of resuspending the protein in water, under precise conditions allowing the subsequent measurement to be maximized.

[0087] The water chosen is preferably osmosis water, but drinking water can also be used.

[0088] 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 the present description, it always includes a variation of + / - 2°C, for example 20°C + / - 2°C or 80°C + / - 2°C.

[0089] A defined quantity of protein is added to said water in order to obtain a suspension titrating 15% + / - 2% in dry matter. To do this, equipment well known to those skilled in the art is used, such as beakers and magnetic bars. A volume of 50 mL is stirred for at least 10 hours at 350 rpm at room temperature. Generally speaking, and unless otherwise indicated, the dry matter contents given in the present description always include a variation of + / - 2%, for example 15% + / - 2%. The pH is adjusted to 7 + / - 0.5 using a pH meter and acid-base reagents, as well known in the prior art.

[0090] 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.

[0091] The following temperature scale is then applied during 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.

[0092] The measurement of the parameter G' is carried out continuously during this scale and is recorded.

[0093] The sixth and final step of test D 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.

[0094] The gelling power is equal to G'2 - G' 1.

[0095] MS rate

[0096] Advantageously, the fava bean protein extract 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 fava bean protein extract. The fava bean protein extract or composition may be in the form of a powder having a particle size d50, which may vary widely, for example from 10 to 500 μm, generally from 50 to 150 μm.

[0097] As will appear below, the fava bean protein extract according to the invention can be obtained by a method comprising a precipitation step at isoelectric pH. A person skilled in the art understands that a fava bean protein extract precipitated at isoelectric pH has a molecular weight profile different from the complete protein contained in the fava bean: in fact, in a fava bean protein extract precipitated at isoelectric pH during isoelectric precipitation, the proteins soluble at isoelectric pH of lower molecular weight are present in very small quantities. In particular, the quantity of albumins in the extract or composition of the invention is generally in trace amounts, and may be, for example, less than 2%, or even less than 1% relative to the protein mass.

[0098] The amounts in each of these ranges of the molecular weight profile, and in particular the preferred ranges, may thus depend in particular on the process conditions, and particularly on the protein precipitation step. The profile may also depend on the solid-liquid separation step of the suspension of precipitated proteins.

[0099] The properties described above can be combined with each other. Thus, different preferred variants of faba bean protein extracts according to the invention are presented below.

[0100] Preferably, the field bean protein extract according to the invention has one or more of the characteristics A to D as indicated in the table below:

[0101] [Tables 1] A Solubility at pH 6 Greater than 50%, preferably ranging from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85% B Solubility at pH 7 greater than 75%, preferably ranging from 75% to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96% C Emulsion capacity greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g D Viscosity less than or equal to 1 Pa.s, advantageously less than or equal to 0.6 Pa.s, for example ranging from 0.05 to 0.6 Pa.s, or from 0.1 to 0.55 Pa.s, for example from 0.2 to 0.5 Pa.s E Gelling power greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa

[0102] In certain embodiments, the faba bean protein extract according to the invention has any of the combinations of the characteristics described in Table 1 above: A, B, C, D, E, A+B, A+C, A+D, A+E, B+C, B+D, B+E, C+D, C+E, D+E, A+B+C, A+B+D, A+B+E, A+C+D, A+C+E, A+D+E, B+C+D, B+C+E, B+D+E, C+D+E, A+B+C+D, A+B+C+E, A+B+D+E, A+C+D+E, B+C+D+E, A+B+C+D+E.

[0103] The fava bean protein extract according to the invention may be in the form of a fava bean protein composition.

[0104] Denatured faba bean protein composition

[0105] Thus, according to a third subject, the invention relates to a denatured fava bean protein composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, said composition having at least one of the following properties: - solubility at pH 6 greater than or equal to 50%, - solubility at pH 7 greater than or equal to 75%, - emulsion capacity greater than 400 mL / g.

[0106] In the present invention, by "denatured fava bean protein composition" or more simply "fava bean protein composition" is meant a composition based on a fava bean protein extract as described in the second subject of the invention. In other words, the fava bean protein composition of the third subject of the invention is a composition mainly comprising a fava bean protein extract as described in the second subject of the invention. Thus, the characteristics in terms of composition or functional properties described above in connection with the denatured fava bean protein extract of the second subject of the invention are applicable to the composition of the third subject of the invention. In certain embodiments, the denatured fava bean protein composition consists of a fava bean protein extract as described in the second subject of the invention.In certain embodiments, the denatured faba bean protein composition is obtained or capable of being obtained by the method of the first subject of the invention.

[0107] The faba bean protein composition may have any of the following combinations of the characteristics described in Table 1 above: A, B, C, A+B, A+C, A+D, A+E, B+C, B+D, B+E, C+D, C+E, D+E, A+B+C, A+B+D, A+B+E, A+C+D, A+C+E, A+D+E, B+C+D, B+C+E, B+D+E, C+D+E, A+B+C+D, A+B+C+E, A+B+D+E, A+C+D+E, B+C+D+E, A+B+C+D+E.

[0108] Certain embodiments of the faba bean protein composition of the invention are described below.

[0109] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0110] In certain embodiments, the fava bean protein composition is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6 preferably measured according to Test A ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85% and a solubility at pH 7, preferably measured according to Test A, ranging from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96%. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0111] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the emulsion capacity of which, preferably determined according to a Test B, is greater than 500 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g. Preferably, the composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0112] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85% and the emulsion capacity of which preferably determined according to a Test B is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0113] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 7, preferably measured according to Test A, ranging from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96% and the emulsion capacity of which, preferably determined according to a Test B, is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0114] In certain embodiments, the faba bean protein composition of the invention is a composition having a faba bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%, the solubility of which at pH 7, preferably measured according to Test A, ranges from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96% and the emulsion capacity of which, preferably determined according to a Test B, is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g. preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0115] In certain embodiments, the faba bean protein composition of the invention is a composition having a faba bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%, the solubility of which at pH 7, preferably measured according to Test A, ranges from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96% and the emulsion capacity of which, preferably determined according to a Test B, is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0116] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%, and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0117] In certain embodiments, the faba bean protein composition of the invention is a composition having a faba bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 7, preferably measured according to Test A, ranging from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96% and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0118] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%, the solubility of which at pH 7, preferably measured according to Test A, ranges from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96% and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0119] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the emulsion capacity of which, preferably determined according to a Test B, is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0120] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 7, preferably measured according to Test A, ranging from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96%, the emulsion capacity of which, preferably determined according to a Test B, is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferentially ranging from 300 to 800 Pa. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0121] In certain embodiments, the fava bean protein composition of the invention is a composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to Test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%, the emulsion capacity of which preferably determined according to a Test B is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa and the gelling power of which gelling agent, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa.Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0122] In some embodiments, the faba bean protein composition of the invention is a composition having a faba bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, the solubility of which at pH 6, preferably measured according to Test A, ranges from 50% to 100%, advantageously ranging from 60 to 90%, for example ranging from 70 to 85%, the solubility of which at pH 7, preferably measured according to Test A, ranges from 75 to 100%, advantageously ranging from 80 to 98%, for example ranging from 85 to 96%, the emulsion capacity of which, preferably determined according to a Test B, is greater than 400 mL / g, advantageously ranging from 500 mL / g to 1000 mL / g, for example ranging from 600 mL / g to 900 mL / g and the gelling power of which, preferably determined according to Test D, is greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa. Preferably, said composition is obtained or capable of being obtained by the process of the first subject of the invention.

[0123] Process for manufacturing a denatured faba bean protein extract

[0124] The method of the first subject of the invention is a method for manufacturing an extract of denatured faba bean protein which includes the following steps: a) preparation of an aqueous suspension of crushed field beans; b) extraction of a protein fraction by solid-liquid separation from the aqueous suspension; c) adjustment to a pH between 4.0 and 5.7 of said protein fraction; d) heat treatment of the protein fraction at a temperature ranging from 50 to 80°C for a time ranging from 1 to 30 seconds to form a suspension of precipitated proteins; e) solid-liquid separation of the suspension of precipitated proteins to recover a field bean protein extract having a protein content by weight relative to its dry matter greater than 70%; f) pH adjustment between 6.3 and 8; g) heat treatment of the fava bean protein extract at pH adjusted to a temperature ranging from 85 to 160°C for a period of time sufficient to obtain denaturation of the fava bean protein; h) possibly drying.

[0125] Step a)

[0126] Step a) comprises the preparation of an aqueous suspension of crushed field beans.

[0127] Generally, ground broad beans are ground broad bean seeds. These seeds typically first undergo steps well known to those skilled in the art, such as cleaning or even the removal of the outer shell of the seed (cellulosic outer envelope), by a well-known step called dehulling. According to the invention, the term "seeds" includes whole broad bean seeds and broad bean cotyledons. The term "seeds" also incorporates whole seeds and damaged cotyledons.

[0128] Before grinding, additional treatments of the grains such as dry heating as described in WO2020 / 260841 may be carried out. Also, wet steeping or wet fermentation of a seed suspension as described in WO2015 / 071498 A1 may be applied.

[0129] In the extraction process, the preparation of a fava bean flour suspension can thus be carried out by suspending in water a fava bean flour obtained by dry grinding of the fava bean seeds. Alternatively, the preparation of a fava bean flour suspension can be carried out by wet grinding. In this case, the fava bean flour suspension can be obtained directly; the water present in the suspension can be retained but can also be renewed.

[0130] The grinding can be carried out by dry grinding, that is to say by passing the previously prepared dry seeds through one or more grinders. Alternatively, the grinding can be wet grinding, that is to say by passing the previously prepared seeds in the form of a suspension of seeds in water through one or more grinders.

[0131] The aqueous solution may be water which may optionally include additives such as in particular antifoam or bacteriostatic compounds.

[0132] According to the variant where the preparation of the seeds is carried out by wet grinding, prior to the grinding step, the aqueous suspension of fava bean may have a ratio by weight of quantity of fava bean / quantity of aqueous solution of between 0.5 and 2. The temperature of the aqueous solution is between 10°C and 80°C. The heating may be carried out using any installation well known to those skilled in the art such as an immersed heat exchanger. Preferably, the temperature is between 25°C and 50°C. When the grinding is carried out by wet grinding, it may be carried out by passing the aqueous suspension of fava bean continuously through one or more grinders to obtain the aqueous suspension of ground fava beans.The grinder(s) may be any type of grinder capable of performing wet grinding, such as wet ball mills, conical wet mills, helical wet mills or wet mills equipped with rotor / stator systems. According to one variant, the grinder may be the one used in the examples of document WO2019 / 053387 in the name of the Applicant. In the variant where the grinder is of the rotor-stator type, this type of grinder may allow continuous grinding by passing the water-bean suspension through said grinder. According to a preferred sub-variant, the method combines two cutting steps (pre-cutting then cutting) using different rotor-stator grinders for each of these cuts. The pre-cutting then the cutting may be carried out one after the other or, alternatively, the cutting may take place. after pre-cutting and then storing the treated water-bean suspension. Such grinders are described in document WO2019 / 158589. Optionally, it is possible to carry out a dilution with water during this step or at the end of this step in order to form the aqueous suspension of ground beans. According to a variant, during grinding, water is added continuously or discontinuously to dilute the aqueous suspension.

[0133] Preferably, at the end of step a), the aqueous suspension of faba bean flour has a dry matter content ranging from 10 to 30%, for example from 15 to 25%.

[0134] Step b)

[0135] Step b) of the process consists of extracting the components of the aqueous suspension of ground broad beans, and in particular extracting a protein fraction by solid-liquid separation of the aqueous suspension of broad beans. According to a variant, before carrying out the solid-liquid separation step, a pH adjustment step of the aqueous suspension of ground broad beans can be carried out. Thus, the solid-liquid separation can take place after adjusting the aqueous suspension of broad beans to a pH ranging from 6 to 9, preferably from 8 to 9, most preferably from 8.5 to 9. This pH adjustment step can be carried out in a stirred tank. This step can be more or less long, and last for example from 1 to 240 minutes, generally from 5 to 60 minutes. To carry out the pH adjustment, it is possible to add any type of acid and / or base, organic or inorganic, or mixtures thereof.Examples of acids include hydrochloric acid, sulfuric acid, ascorbic acid, citric acid, or mixtures thereof. Examples of bases include sodium hydroxide, potassium hydroxide, or lime, and mixtures thereof. This addition of base or acid, as well as the pH measurement, can be done online. The base and / or acid can be in the form of aqueous solutions.

[0136] Generally, the protein fraction is the soluble part of the aqueous suspension and the starch and fiber rich fraction is the insoluble part.

[0137] Thus, in certain embodiments, step b) of extracting a protein fraction comprises - the solid-liquid separation of the aqueous suspension from step a), and - the recovery of the liquid fraction, which is preferably an aqueous phase.

[0138] The solid-liquid separation can in particular be carried out by means of at least one separation step with a decanter, in particular a centrifugal decanter, a centrifuge or even with hydrocyclones.

[0139] The recovery of the liquid phase can be done by any method known to those skilled in the art, typically by overflow, by decantation, or using a sieve or a filter.

[0140] The protein fraction obtained at the end of step b) is typically presented in the form aqueous suspension form, preferably having a pH between 6 and 9, preferably 8 to 9, most preferably 8.5 to 9. The dry matter of the protein fraction can vary, and for example range from 3 to 15%.

[0141] It is also possible to separate more than two insoluble fractions, and for example to recover a first insoluble fraction richer in starch and a second insoluble fraction richer in fibers.

[0142] Thus, according to a variant of the process, a starch-rich fraction and / or a fiber-rich fraction is recovered from the insoluble part resulting from the solid-liquid separation step b). By starch-rich fraction and fiber-rich fraction, is generally meant a fraction comprising at least 50% starch or fiber. The methods for quantifying starch and fiber are known to those skilled in the art and specific methods are indicated later in the description. These fractions are recovered conventionally by known separation methods. The process can also make it possible to recover one or more fractions enriched in fibers and / or starch, which are removed from the suspension, and to recover the protein fraction useful following the process of the first subject of the invention.

[0143] Step cl

[0144] The method also comprises a step c) of adjusting said protein fraction to the pH, which may be the isoelectric pH of the protein. Isoelectric pH means a pH close to which the net electrical charge of the protein of the protein fraction is zero. The pH may be adjusted to a pH between 4.0 and 5.7, preferably between 4.2 and 5.7, or even between 4.6 and 5.0. The pH rectification may be carried out by adding acid, organic or inorganic, for example hydrochloric acid, sulfuric acid, ascorbic acid or citric acid and mixtures thereof. This step c) may be carried out in a tank, stirred or not. It may be more or less long, and last for example from 1 to 240 minutes, generally from 5 to 60 minutes. This addition of acid as well as the measurement of the pH may be carried out online and the acid may be in the form of an aqueous solution.

[0145] Step d)

[0146] The method also comprises a step of heat treatment d) of the protein fraction at the adjusted pH. This step comprises a stage of heating the suspension of precipitated proteins. This stage is carried out at a temperature ranging from 50 to 80°C to form a suspension of precipitated proteins. It can be carried out for a duration ranging from 1 to 30 seconds, preferably from 1 to 20 seconds, most preferably from 1 to 10 seconds. To carry out this heating, a heat exchanger is generally used. It can be of the type according to the principle of indirect heating or according to the principle of direct heating, generally by steam injection. Preferably, the heating is carried out by steam injection. Advantageously, step heat treatment step d) comprises a heating stage followed by a cooling stage of the suspension of precipitated proteins. In the variant where the heat treatment step d) comprises, following the heating stage of the suspension of precipitated proteins, a cooling stage of said suspension, this cooling stage is preferably obtained by rapid cooling known as "flash-cooling", leading to immediate cooling. At the end of this stage, the temperature can range from 60 to 75°C, for example between 64 and 70°C. This flash-cooling is carried out by applying a vacuum to the suspension of precipitated proteins, the vacuum applied being determined according to the cooling temperature chosen.

[0147] Step e)

[0148] In the remainder of the process, a fava bean protein extract is separated from the suspension of proteins precipitated in step d). This solid-liquid separation can be carried out using the means indicated for the separation means indicated in step b). The fava bean protein formed during this step mainly comprises proteins insoluble at isoelectric pH which are separated from the liquid fraction. In the liquid fraction are generally found other proteins soluble at isoelectric pH, albumins but also soluble carbohydrates. The recovered solid fraction comprises the fava bean protein and is generally a concentrated aqueous suspension of fava bean protein.

[0149] Thus in certain embodiments, step e) of solid-liquid separation of the suspension of precipitated proteins comprises: - the solid-liquid separation of the precipitated protein suspension from step d), and - recovery of the solid fraction,

[0150] The protein fraction obtained at the end of step e) is typically in the form of a solid fraction, which is preferably a concentrated aqueous suspension of fava bean protein. The recovered solid fraction has a dry matter content which generally ranges from 25 to 50%, or from 30 to 40%. The mass composition of the fava bean protein extract can vary and will generally comprise mainly proteins other than albumins (in particular in the form of globulins) but also starch, lipids, fibers, and / or sugars. This solid fraction can be diluted by adding water to be more easily handled in the following steps.

[0151] Step D

[0152] At the end of this step e), the method comprises a step f) of adjusting the pH of the faba bean protein extract to a pH between 6.3 and 8, generally from 6.5 to 7.5. This step can be carried out by adding an inorganic or organic base, for example by adding soda, lime, potash or a mixture of these. Raising the pH is generally done by adding a basic aqueous solution.

[0153] Step g)

[0154] The method further comprises a step g) of heat treatment of the fava bean protein extract at the adjusted pH at a temperature ranging from 85 to 160°C for a time allowing denaturation of the fava bean protein and preservation of solubility. The temperature and time conditions can vary widely in this step, for example ranging from 85 to 140°C and lasting from 0.1 seconds to several minutes. The step is carried out so that, at the end of the extraction process, the fava bean protein extract can exhibit the solubility and denaturation characteristics (including enthalpy) set out previously in the description. According to a first variant of this additional heat treatment step, the temperature ranges from 85 to 100°C and its duration ranges from 5 seconds to 5 minutes. According to a second variant of this additional heat treatment step, the temperature ranges from 100 to 110°C and its duration ranges from 1 second to 4 minutes.According to another preferred variant, this heat treatment step is carried out at a temperature ranging from 110 to 150°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 heat treatment step g) makes it possible to sanitize the faba bean protein extract in order to make it suitable for direct use in the manufacture of food or pharmaceutical products. To carry out this heat treatment step, the faba bean protein extract may be in the form of an aqueous dispersion, preferably having a dry matter ranging from 10 to 25%, for example from 15 to 20%.

[0155] Advantageously, the method of the first subject of the invention comprises, following the heat treatment step g), a step of cooling the faba bean protein extract. According to a preferred variant, this cooling step is obtained by vacuum cooling (in English, flash-cooling). This flash-cooling step is carried out by applying a vacuum to the aqueous dispersion of the faba bean protein extract, the vacuum applied being determined according to the cooling temperature chosen. At the end of this step, the temperature can vary according to the temperature of the step preceding it (the heat treatment step g)). 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 can range from 30 to 100°C.For example, in the advantageous mode where the heat treatment temperature ranges from 110 to 150°C, the cooling temperature may range from 60 to 100°C, for example may be around 80°C. Based on the above and the following examples, the person skilled in the art will be able to select the conditions. allowing to obtain the desired functional characteristics as well as the denaturation of the protein.

[0156] The solubility of a protein can depend on many parameters of its production process. As is known from the prior art, the heat treatment steps lead to a denaturation of the protein, which leads to a loss of functionality, and in particular a reduction in solubility. This same prior art describes faba bean proteins of high solubility but which are not or only slightly heat treated, which leads to the provision of undenatured proteins. However, these methods do not allow the microbial load in the protein to be reliably controlled, which makes these methods not industrially exploitable. On the contrary, by the method of the first subject of the invention, it was possible to provide a new extract of faba bean protein which makes it possible to overcome these problems.This process ensures that the faba bean protein extract has a very low microbial load, allowing it to be used directly in food or pharmaceutical products, without even an additional heat treatment step. Thus, by selecting the process conditions, the process led to the denaturation of the protein but while surprisingly providing excellent solubility at neutral pH, or even at slightly acidic pH (pH 6).

[0157] Shearing step

[0158] According to a variant of the method, it comprises a step of shearing the faba bean protein extract, 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.

[0159] The shearing step can take place before or after the heat treatment and / or pH raising steps.

[0160] Homogenization step

[0161] According to another variant, the method comprises a step of homogenization of the field bean protein extract.

[0162] To carry out this homogenization step, any type of homogenizer can be used. According to the invention, it is understood to mean equipment comprising a high-pressure pump and a homogenization head in which the equipment is designed so that the product to be homogenized passes under pressure through this homogenization head. A homogenization head consists of a reduced orifice generally comprising a seat, a valve and a shock ring. The passage of the aqueous dispersion of faba bean proteins through the homogenizer can thus enable the homogenization of the faba bean protein extract. The homogenization may be low-pressure homogenization, high-pressure homogenization, or ultra-high-pressure homogenization. The homogenization pressure may vary widely and, depending on the homogenization technique used, may range from 1 to 1000 bar, for example, from 20 to 800 bar. According to one variant, the homogenization pressure ranges from 20 to 200 bar, for example, from 50 to 150 bar. According to another variant, the homogenization pressure ranges from 200 to 800 bar, for example, from 300 to 800 bar. According to one variant, the homogenization is single-effect homogenization. According to another variant, the homogenization is multiple-effect homogenization, for example, double-effect homogenization. Homogenizers that can be used are sold, for example, by GEA or Tetra Pak.

[0163] The homogenization step can take place before or after the heat treatment and / or pH raising steps.

[0164] Drying step

[0165] The method according to the invention may also comprise a step of drying the faba bean protein extract. 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, for example, 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.

[0166] The method generally does not include any membrane protein separation step. These techniques are well known to those skilled in the art and mainly include ultrafiltration, diafiltration, reverse osmosis and nanofiltration. Membrane separation techniques, in particular ultrafiltration, make it possible to fractionate the proteins in a composition.

[0167] Enzymatic modification step

[0168] The faba bean protein extract of the invention can then be enzymatically modified. By enzymatically modified protein, the person skilled in the art means a protein whose protein structure has been deliberately modified by the addition to the protein of at least one enzyme capable of modifying the protein structure. This enzyme can be chosen from proteases, peptidases, deamidation enzymes, for example those of type EC 3.5.1 such as glutaminase or deimination enzymes, for example those of type EC 3.5.3 such as peptidylarginine deiminase. These protein modification enzymes are known to modify the physicochemical and / or organoleptic properties of the protein. For example, it is known to document WO2019 / 233920 A1 that peptidylarginine deiminase makes it possible to reduce astringency, in particular the astringency of rapeseed protein. The faba bean protein extract of the invention is not enzymatically modified. An advantage of the invention is that it is possible to modify the organoleptic properties of the faba bean protein extract, and in particular to give it excellent solubility, without even needing to enzymatically modify the protein.

[0169] Textured composition of fava bean protein

[0170] According to a fourth subject, the invention relates to a textured fava bean protein composition obtained by texturizing a fava bean protein extract according to the first subject of the invention or a fava bean protein composition according to the second subject of the invention.

[0171] By "textured fava bean protein composition" is meant in the present invention a composition comprising a fava bean protein which has been subjected to a physical and / or chemical process aimed at modifying the protein in order to give it a specific ordered structure. In the context of the present invention, the texturizing of the fava bean protein aims to give it the appearance of fibers such as those present in animal meats.

[0172] In certain preferred embodiments, the textured fava bean protein composition is characterized in that it is obtained by wet texturizing a fava bean protein extract according to the first subject of the invention or a fava bean protein composition according to the second subject of the invention.

[0173] By "wet texturing" is meant in the present invention a texturing process, in particular by cooking-extrusion, in which the amount of water in the mixture present in the extruder represents more than 40% of the total weight of the ingredients used during the process, preferably between 40% and 90%. Typically, as detailed below, the textured faba bean protein composition of the present invention is preferably prepared by cooking-extrusion by introducing a powder and water into an extruder, said powder containing a faba bean protein extract according to the second subject of the invention or a composition according to the third subject of the invention, and optionally a legume fiber, typically a pea fiber, and / or starch, typically potato starch.

[0174] Preferably, the extruded fava bean protein composition is obtained by wet extrusion of a powder containing only a fava bean protein extract according to the first subject of the invention or a composition according to the second subject of the invention.

[0175] Use of fava bean protein extract

[0176] The fifth subject of the invention is the use of the field bean protein extract. of the second subject of the invention, of the denatured fava bean protein composition of the third subject of the invention, or of the textured fava bean protein composition of the fourth subject of the invention, for the manufacture of food or beverage products, in particular plant-based alternatives to milk.

[0177] In the following, unless otherwise stated, the terms "fava bean protein extract of the invention", "fava bean protein of the invention" and "protein of the invention" encompass the fava bean protein extract of the second subject of the invention, the denatured fava bean protein composition of the third subject of the invention, as well as the textured fava bean protein composition of the fourth subject of the invention.

[0178] Generally, the faba bean protein extract 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.

[0179] A particularly interesting use of the protein of the invention concerns its use in drinks which have a pleasant taste.

[0180] In the beverages, the protein content of the invention can vary widely and can also be a high protein drink. The amount of protein of the invention 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 alternatives to milk or milk substitutes, including barista-type milks or coffee creamers.It may also include other ready-to-drink beverages, whether acidic or not, such as carbonated beverages (including, but not limited to, carbonated soft drinks), non-carbonated beverages (including, but not limited to, non-carbonated soft drinks such as flavored waters, fruit juices, and sweetened or unsweetened tea or coffee-based beverages), alcoholic beverages such as beers or spirits, smoothies, beverage concentrates (including, but not limited to, liquid concentrates and syrups, and non-liquid "concentrates," such as freeze-dried and / or powdered preparations or "powder mixes").

[0181] 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 said bread products; sweet bakery products (including, but not limited to, rolls, cakes, pies, pastries, waffles, crepes, muffins, pancakes, and cookies); 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 products and the like, such as cream fillings);snack bars (including, but not limited to, energy, cereal, nut, and / or fruit bars).;

[0182] These may also be gelled desserts such as puddings or custards 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 ice cream and all other types of ice cream - and frozen non-dairy desserts such as non-dairy ice cream, sorbet and others).

[0183] Other products conventionally prepared from animal milk may also include the faba bean protein extract 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 "Greek" or "skyr" type yogurts which are high-protein yogurts (often ranging from 8 to 20 g protein) as well as white cheeses and petits suisses.It can also be cheese substitutes such as spreadable, processed, cooked and uncooked pressed cheeses, soft cheeses, spun cheeses, blue cheeses; it can be 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.

[0184] Other products that may include the faba bean protein extract of the invention are also sauces such as salad dressings or mayonnaise or ketchup based sauces or syrups.

[0185] Also, the fava bean protein extracts of the invention may be incorporated in 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 include sweetened spreads (including, but not limited to, jellies, jams, nut butters such as peanut butter, spreads, and other spreadable products).

[0186] The faba bean proteins of the invention can also be used as a carrier or in flavor encapsulation.

[0187] Other types of foods and beverages not mentioned herein but which conventionally comprise one or more proteins and may also be contemplated within the scope of the present invention. In particular, animal foods (such as pet foods) are explicitly contemplated.

[0188] The fava bean protein extract can also be used, optionally after texturizing, in meat substitutes such as emulsified sausages or hamburgers, or fish or seafood substitutes. It can also be used in egg replacement formulations or for the manufacture of protein products such as tofu or tempeh. Textured proteins generally mean proteins textured by extrusion, i.e. in particular dry extrusion ("dry extrusion" or "Textured Vegetable Protein"), wet extrusion ("high moisture extrusion"). The extruders can be single-screw, twin-screw or multiple-screw extruders. In the case of twin-screw extrusion, the extrusion can be co-rotating or counter-rotating. Examples of multiple-screw extrusion include the planetary extruder ("planetary extruder") or the ring extruder ("ring-extruder").Other more specific technologies such as shear cell technology, microextrusion or 3D printing can also be mentioned. As demonstrated in the Examples section, the proteins of the invention are particularly interesting for the manufacture of textured proteins by wet extrusion.

[0189] The food products or beverages may in particular be used in specialized nutrition, for example for specific populations, for example for babies or infants, children, adolescents, adults, the elderly, athletes, people suffering from an illness. These may be meal replacement nutritional formulas, complete nutritional drinks, for example for weight management or in clinical nutrition (for example tube feeding or enteral nutrition).

[0190] Fava bean protein extract can be used as the sole source of protein, but can also be used in combination with other additional proteins, plant or animal. These additional proteins can be hydrolyzed or non-hydrolyzed. Generally, these additional proteins are in the form of concentrates or isolates. 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 mixtures, chosen from the same family or from different families. By "legume", we generally mean the family of dicotyledonous plants of the order Fabales.Several legumes are important cultivated plants including soybeans, beans including mung beans, chickpeas, peas, 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 faba bean protein other than the extract 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. Examples of wheat proteins include NUTRALYS® W marketed by the Holder. Examples of rice proteins include NUTRALYS® RICE 800XF and NUTRALYS® RICE 850XF. Tubers include carrots, cassava, konjac, potatoes, Jerusalem artichokes, and sweet potatoes.Oilseed plants are generally plants that produce seeds from which oil is extracted. Oilseed plants can be chosen from sunflower, rapeseed, peanut, sesame, pumpkin, or flax. Animal proteins can be, for example, egg or milk proteins, such as whey protein, casein, or caseinates. The fava bean protein extract can be used with another of the above-mentioned protein sources, in particular pea, rice or wheat protein, in the following protein mass ratios of fava bean protein / other protein: 1 / 99, 2 / 98, 3 / 97, 4 / 96, 5 / 95, 6 / 94, 7 / 93, 8 / 92, 9 / 91, 10 / 90, 11 / 89, 12 / 88, 13 / 87, 14 / 86..., 86 / 14, 87 / 13, 88 / 12, 89 / 11, 90 / 10, 91 / 9, 92 / 8, 93 / 7, 94 / 6, 95 / 5, 96 / 4, 97 / 3, 98 / 2, 99 / 1.This mass ratio may advantageously be from 10 / 90 to 90 / 10, or from 25 / 75 to 75 / 25, or from 35 / 65 to 65 / 35, all of these ranges being able to be combined with each other or with the ratios described above. In the examples below, the faba bean protein of the invention was used with the pea protein according to the ratio ranges above. The other proteins may also be used with the faba bean protein extract of the invention according to the same ratio ranges. The faba bean protein extract of the invention may also 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 for . improve the PDCAAS of the protein portion of the food product or to provide other functionalities. The aforementioned cereal proteins, particularly those from rice or wheat, are good choices for improving the PDCAAS, especially when the dry weight ratio of faba bean protein to cereal protein ranges from 85 / 15 to 65 / 35.

[0191] The fava bean protein extract 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.

[0192] 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

[0193] Methods

[0194] For the determination of the viscosity profile in water, the measurements are carried out on an aqueous solution of faba bean protein extracts at 15% dry matter (osmosed and azidated water at 200 ppm to prevent any bacteriological risk), in an AR2000 rheometer from the company TA Instruments, having a concentric cylinder geometry, with a shear rate of 0.6 103 at 600 s 1 in 3 minutes (log) and at a temperature of 20°C (3 min of temperature equilibrium before testing).

[0195] 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 in the Table.

[0196] Enthalpy of denaturation

[0197] The enthalpy by integration of the endothermic peak was quantified using the calorimeter control software. The start of denaturation is expressed by the Tonset (°C) and the end of Tendset (°C). The critical denaturation temperature Tpeak (°C) is determined at the top of the peak.

[0198] Total lipids (%)

[0199] The total lipid content can be determined by hydrolysis with boiling hydrochloric acid, and precipitation of the total lipids by cooling. Separation by filtration, drying and isolation of the lipids by extraction with hexane.

[0200] Operating mode:

[0201] Weigh a test portion accurately to within 0.001 g of a quantity of sample chosen so as to have approximately between 100 and 500 mg of lipids in this test portion. In other words, if the quantity of lipids is approximately 10%, the weight of the sample must be between 1 and 5 g.

[0202] Place the test sample in a 250 mL flask. Add 100 mL of chlorinated acid water, a few fragments of pumice stone and heat under reflux for 60 minutes. Cool to precipitate the lipids and add a sufficient quantity of filter aid to avoid any loss of fat.

[0203] Quantitatively filter the mixture through a wet double filter. Rinse the flask several times with cold distilled water. Wash the residue with cold distilled water until the filtrate is neutral (check with pH indicator paper). Dry the double filter containing the residue overnight under a hood and finish drying in the oven for 6 to 7 hours.

[0204] Extraction of total lipids and removal of solvent

[0205] Place the double filter and its contents in the extraction cartridge. Cover with a piece of cotton wool and insert it into the extractor. Pour 200 mL of Hexane into the flask, assemble the flask - extractor - condenser and heat under reflux for 6 hours.

[0206] After cooling, the hexane contained in the Soxhlet is recovered in the flask. Set up the flask - solvent recovery - condenser, close the tap and heat moderately to recover the hexane by distillation.

[0207] Dry the residue by placing the flask or beaker in the oven for 1 hour 30 minutes. Cool in a desiccator and weigh. Dry again for 30 minutes to ensure that the weight of the fat remains constant (the weight loss between two successive weighings must be less than 1 mg)

[0208] Example 1: Field bean protein according to the invention

[0209] Approximately 900 kg of fava beans were used. The outer fibers of the fava beans were first separated from the seeds by crushing (mechanical separation of the outer shell and the fava bean seed) and dehulling. Water at 45°C was used in this first part of the process: fava bean seeds and water were continuously fed into a Bruynooghe brand submerged screw blancher. The fava bean / water weight ratio was approximately 0.15. The water inlet was made entirely at the entrance to the blancher and the fava beans were immediately introduced into the water. The screw speed is adjusted so that the fava beans pass through the blancher in approximately 3 minutes. The water-fava bean suspension was immediately ground by continuous wet grinding to obtain a suspension of ground fava bean of approximately 13.5% dry matter.The suspension of ground broad beans was adjusted to pH 8.7 by adding, continuously and in-line, an aqueous alkaline mixture of sodium hydroxide (10% of the solution by mass of the mixture) and potassium hydroxide (10% of the solution by mass of the mixture). The suspension of ground broad beans was cooled to approximately 8°C by passing through a plate heat exchanger and then transferred to a stirred storage tank. This suspension fed a centrifugal decanter (Flottweg Z3). The protein fraction was recovered in the overflow (approximately 6% of material). The protein fraction was adjusted to pH 4.95 in a stirred tank by adding 4.5 kg of pure ascorbic acid and hydrochloric acid until the desired pH was obtained. The protein fraction was then heat-treated by injecting steam at 72°C into a GEA skid for approximately 5 seconds, after an initial immediate preheating step by passing it through a plate exchanger. The protein fraction was then immediately cooled by flash cooling to 64°C. Immediately, the heat-treated protein fraction was passed through a Flottweg Z3 decanter centrifuge. The recovered protein sediment (the "underflow") was diluted in hot water (60°C) so that it could be pumped. This sediment was then immediately adjusted to a dry matter content of approximately 15% and then rectified to pH 6.8 with the aqueous alkaline mixture.The faba bean protein floc was heat treated at 140°C for 5 seconds and then flash cooled to approximately 75°C. This faba bean protein floc was passed through a high-pressure homogenizer at 200 bars and then atomized in a TGE nozzle atomizer (inlet temperature 200°C, outlet temperature 60°C). The recovered faba bean protein powder (hereinafter Faba bean protein Ex.l or more simply Ex.l) was then analyzed.

[0210] Example 2: Field bean protein according to the invention

[0211] Example 2 is identical to Example 1 except that the protein fraction was adjusted to pH 4.75 with the mixture of ascorbic acid and hydrochloric acid instead of 4.95. Furthermore, before the heat treatment step at 140°C for 5 seconds, the sediment was rectified to pH 6.5 instead of 6.8 with the aqueous alkaline mixture. The recovered fava bean protein powder (Fava bean protein Ex.2 or more simply Ex.2) was analyzed.

[0212] The results of the analyses are presented in the table below:

[0213] [Tables2] Ex.l Ex.2 T FULL BUNGE SHUAN GTA FOOD COSUN NUTRA LYS S85F NUTRA LYS F85 M Protein (%) in dry mass 94.8 93 94.2 88 92 91.2 83.8 83.8 Total lipids (%) 6.8 6.9 7 6.6 7.6 5.7 9.2 9.2 Dry mass (%) 95.1 95.2 92.8 95.6 92.4 93.8 92 92.1 Solubility (%) pH 3 100 100 22 49 NM 100 NM NM Solubility (%) pH 6 80 82 13 33 NM 17 NM NM Solubility (%) pH 7 94 90 18 51 28 95 42 75 Emulsion capacity (mL oil / g) 700 650 250 300 450 150 400 NM Gelling power (Pa) 174 720 533 603 125 <50 109 206 Viscosity 15% (40s1 ) 0.3 0.5 2.6 0.7 0.6 0.1 NM 0.2 Enthalpy (J / g) 0 0 0 0 0 nd 0 0

[0214] The protein extracts were also tested in comparison with commercial proteins in different applications which will be described below and which demonstrate the interest of the faba bean protein extracts of the invention, in particular in food and beverage products.

[0215] Example 3: Evaluation of faba bean protein for the manufacture of alternatives to milk-based food or beverage products.

[0216] Alternatives to milk

[0217] Milk alternatives have been manufactured so that the protein level in the formulation is equal to 5% of the total weight.

[0218] Thus the following formulations without emulsifier of milk alternatives are produced and reported in the Table below (proportions by weight of the milk alternative recipe):

[0219] [Tables3] Milk alternative Inv.l CPI CP2 Protein source Ex.l T-Full NUTRALYS S85 F protein quantity 5.60 5.78 6.40 Powdered cane sugar 2 2 2 Sunflower oil 1.50 1.50 1.50 Gellan gum 0.12 0.12 0.12 Demineralized water qsp qsp qsp Total 100 100 100

[0220] The formulations are prepared by the following process: 1) Heat the oil in a bain-marie (65°C) 2) Dry mixing of solid ingredients 3) Heat the water to 70°C 4) Place the solid ingredients with the water in a Silverson mixer and mix for 30 minutes at 2500 rpm. 5) Then set the mixer to 5000 rpm and add the oil in a continuous stream for one minute 6) UHT sterilization of the formulation: 142°C for 5 seconds 7) Homogenization at 75°C 1st stage 170 bars, 2nd stage 30 bars 8) Cool to 4°C and bottle and store at 4°C

[0221] It has been observed that the fava bean protein of Example 1 according to the invention makes it possible to provide a milk alternative that is much more stable to phase shift than the milk alternative made from commercial fava bean protein. This milk alternative is even slightly more stable than the milk alternative made from commercial pea protein.

[0222] Study of foaming power

[0223] In order to study the interest of the protein of the invention in barista milk type applications, the foaming power of the milk alternatives was determined according to the following method: 1) Place 150 mL of milk alternative to froth into the Nespresso® Aeroccino 3. 2) Start the appliance in heating mode for one cycle, the milk alternative foam reaching a temperature of 68°C 3) Recover the foamy product and immediately determine the volumes of foam and liquid obtained

[0224] The results are presented in the Table below:

[0225] [Tables4] Milk alternative Inv.l CPI CP2 Foam volume 230 22 202 Liquid volume 20 138 48

[0226] This evaluation demonstrates that the faba bean protein of Example 1 according to the invention makes it possible to provide a milk alternative that is much more foamed than the milk alternative made from commercial faba bean protein. This milk alternative is even slightly more foamed than the milk alternative made from commercial pea protein.

[0227] Yogurt without texturizer

[0228] Yogurt formulations comprising 3.5% protein and free from texturizer were made from the following recipes, the quantities being expressed in total weight of the yogurt:

[0229] [Tables5] Yogurt Inv.l Inv.2 CP Protein Ex.l Ex.2 NUTRALYS® S85 F Demineralized water 89.17 89.1 88.62 Powdered cane sugar 4.2 4.2 4.2 Sunflower oil 2.75 2.75 2.75 Protein isolate 3.88 3.95 4.43 Lactic culture Vega harmony qsp qsp qsp

[0230] Yogurts are made using the following process: 1) Heat demineralized water to 55°C 2) Add the protein isolate with moderate stirring (480 rpm) and hydrate at 55°C for 20 minutes 3) Add the sugar and continue mixing for 2 minutes. Add the oil while stirring, increasing the shear (1800 rpm) and mix for 5 minutes to obtain an emulsion. 4) Place the emulsion in a NIRO homogenizer (150 bar 1st stage, 45 bar 2nd stage, 60°C) 5) Pasteurize the homogenized emulsion at 95°C for 10 minutes (Hotmix) 6) Cool to 42°C then add the lactic cultures 7) Maintain at 42°C until the pH is 4.60

[0231] The table below shows the hardness of the yogurt after 7 days.

[0232] [Tableauxô] Yogurt Ex.l Ex.2 CPI Hardness after 7 days (g) 31.4 43.3 <25

[0233] Yogurts with texturizers are also prepared without difficulty from the fava bean proteins of the invention.

[0234] Ice cream

[0235] Ice cream formulations comprising approximately 2.5% protein were made according to the following recipes:

[0236] [Tables7] Ice Cream Inv CP Protein Ex.2 TFULL Demineralized Water 65.44 65.4 Hydrogenated Coconut Oil 8 8 Powdered Sugar 12 12 Glucose Syrup 6080 (ARUGULA) 11.5 11.5 Cremodan SE 30 (stabilizer, emulsifier) ​​0.25 0.25 Protein Isolate 2.81 2.85

[0237] 17 L of ice cream were made using the following process: 1) Heat the water to 70°C in the Silverson mixer 2) Add the glucose syrup and three-quarters of the sucrose. Heat to 60°C and mix for 5 minutes at 2000 rpm. 3) Add the stabilizing emulsifying system and the remaining sugar. Maintain at 60°C and mix for 5 minutes at 2000 rpm. 4) Add the oil and create a pre-emulsion by mixing at 6000 rpm for 5 minutes then reduce agitation to 2000 rpm and mix for 15 minutes 5) Heat to 70°C and homogenize in a Powerpoint homogenizer (200 bar 1st stage, 60 bar 2nd stage) 6) Pasteurize at 80°C for 3 minutes through the Powerpoint tubular heat exchanger 7) Cool to 4°C 8) Mature statically overnight 9) Cooling with a TetraPak continuous freezer 10) Freeze for 3 hours at -22°C 11) Store in the freezer at -18°C

[0238] The ice creams were tasted by a trained panel. With regard to the creaminess of the ice cream, a tendency towards more creaminess for the ice cream prepared with the proteins of Example 2 of the invention was detected. With regard to taste, significant differences (sweeter, less bitter and less vegetal) in favor of the ice cream of the invention are observed in comparison with the comparative ice cream, with in addition for the ice cream of the invention caramel, vanilla and milky notes.

[0239] Example 4: Evaluation of the field bean protein according to the invention in extrusion

[0240] Evaluation of the faba bean protein of Example 1 in wet extrusion

[0241] A mixture of powders is made according to the following recipes described in the Table below, expressed in mass of the ingredients:

[0242] [Tables8] Ingredients Inv.l CP.l Inv.2 Inv.3 Fava bean protein Ex. 1 92 0 100 46 NUTRALYS® F85M 0 92 0 46 Pea fiber ROQUETTE® I50M 5 5 0 5 Potato starch 3 3 0 3 Extrusion temperature profile 2 1 1 1

[0243] The proteins tested are the faba bean protein of example 1 as well as the commercial pea protein NUTRALYS® F85M.

[0244] This mixture is introduced by gravity into a LEISTRITZ ZSE 27MAXX extruder from the company LEISTRITZ.

[0245] The mixture is introduced with 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%.

[0246] 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 of which the 2nd module is thermoregulated at 30°C; The extrusion screw is rotated at a speed equal to 350 rpm and sends the mixture into the die, except in the case of test Inv.2 during which a speed of 800 rpm was used. 2 temperature profiles were used according to the test.

[0247] The temperature profile 1 of the extruder, equipped with 15 heatable barrels, is detailed below:

[0248] [Tables9] Z15 Z14 Z13 Z12 Zll Z10 Z9 Z8 Z7 Z6 Z5 Z4 Z3 Z2 ZI T°C profile 90 120 120 130 115 115 110 90 60 60 60 60 35 35

[0249] Extruder temperature profile 2 is detailed below:

[0250] [TableauxlO] Z15 Z14 Z13 Z12 Zll Z10 Z9 Z8 Z7 Z6 Z5 Z4 Z3 Z2 ZI T°C profile 120 120 130 150 150 130 100 90 80 60 60 60 35 35

[0251] 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).

[0252] For the 4 tests, the extrusion parameters are reported below:

[0253] [Tableauxll] Mixture Inv.l CP.l Inv.2 Inv.3 Profile Temperature 2 1 1 1 Torque (%) 12.9 13.8 17 13 Pressure (bar) 14.4 32 24.7 24 T°C material (°C) 114 92 101 60

[0254] Observation of the fibration of the bands

[0255] To observe the fibration of the strip, the strip is cut in two 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 [Fig.l] to [Fig.4]. Furthermore, depending on the fibration observed, the test is rated from 1 / 5 (no fibration of the strip) to 5 / 5 (strong fibration of the strip).

[0256] Chopping test

[0257] To carry out this test, the strip is deep-frozen and then frozen at -18°C. Then, 140g of the strip is taken and then tempered at 15-17°C before being placed in a Stephan UMC 5 mixer equipped with a serrated blade. The mixer was activated for 10 sec at 1500 rpm and then 70g of the sample were analyzed. The pieces obtained are observed (Figures [Fig.5] to [Fig.8]). Depending on the size of the pieces, the test is rated 1 / 5 (poor resistance to tearing and presence of numerous small pieces and / or very large pieces) to 5 / 5 (good resistance to tearing and absence of small and very large pieces). As, very generally in the field of meat substitutes, the textured protein strip is not consumed as is but is cut before use for the production of a substitute, the resistance to tearing makes it possible to quantify the capacity of the textured protein strip to be cut while retaining medium-sized pieces, in order to contribute to the texture of the final product.

[0258] Results

[0259] The scores for both tests are shown in the Table below.

[0260] [Tablesl2] Mixture Inv.l CP.l Inv.2 Inv.3 Note dilaceration 4 2 3 4 Note fibration 5 4 4 5

[0261] [Fig. 1] shows a wet extrusion strip obtained from a mixture Inv.l comprising the faba bean protein of Example 1 combined with pea fibers and potato starch.

[0262] [Fig.2] shows a wet extrusion strip obtained from a CP.1 mixture comprising a commercial pea protein.

[0263] [Fig.3] shows a wet extrusion strip obtained from a mixture Inv.2 comprising only the faba bean protein of Example 1.

[0264] [Fig.4] shows a wet extrusion strip obtained from the faba bean protein of Example 1 in combination with comparative pea protein.

[0265] [Fig.5] shows the results of the tearing test of a wet extrusion strip obtained from a mixture Inv.l comprising the faba bean protein of Example 1 combined with pea fibers and potato starch.

[0266] [Fig.6] shows the results of the tearing test of a wet extrusion strip obtained from a CP.1 mixture comprising a commercial pea protein.

[0267] [Fig.7] shows the results of the tearing test of a wet extrusion strip obtained from a mixture Inv.2 comprising only the faba bean protein of Example 1

[0268] [Fig.8] shows the results of the tearing test of a wet extrusion strip obtained using a mixture Inv.3 comprising the faba bean protein of Example 1 in combination with commercial pea protein.

[0269] Observation of figures [Fig.l] and [Fig.2] shows good fibration of the strip obtained with the mixture of starch and pea fiber with the field bean protein. (Inv.l), which is slightly improved compared to that obtained with pea protein (CP.l).

[0270] Observation of [Fig.3] also shows that, even when the recipe does not include fiber or starch (Inv.2), the textured faba bean protein strips showed good fibration even without the addition of fiber.

[0271] After the dilaceration test, it was observed that the textured faba bean proteins have fewer fine particles than the textured pea proteins, making them more easily transformable and allowing final products with a different texture to be obtained from the textured pea proteins, without the need to add other ingredients.

[0272] Furthermore, the Inv.3 strip, combining pea protein and faba bean protein, has less hardness than the Inv.1 strip, which could make it possible to obtain a finished product with a more pleasant texture.

[0273] Thus, whatever the formulation, by partially or totally replacing the pea protein with the field bean protein of the invention, it was possible to provide strips with improved properties.

[0274] Example 5: Use of field bean protein extract in sausage emulsified

[0275] To determine whether the faba bean protein extract is suitable for use in the manufacture of an emulsified sausage, the following model system is used:

[0276] In a Stephan UMC-5, 500 grams of water and 100 grams of protein were weighed, mixed at 5°C at 3000 rpm with the emulsion blade for 2 minutes under vacuum. By suction, 500 grams of sunflower oil were added and continued stirring for 2 minutes and 30 seconds, then 22 grams of table salt were added and mixed for 30 seconds. The mixture was placed in a 142 mL cylindrical can (diameter 83 x height 44 mm) filled to three-quarters and then sealed tightly.

[0277] Since food products are generally produced using processes including pasteurization or sterilization, these two heat treatments of these emulsions are carried out.

[0278] Pasteurization:

[0279] The can is placed in a water bath at 75°C for 1 hour 30 minutes

[0280] Sterilization:

[0281] The can is placed in a sterilizer at 115°C for 1 hour. After storage at 4°C overnight, the can is immediately opened and tested by texturometry.

[0282] Texturometry:

[0283] The texture is analyzed with a TA-HDPLUS (Stable Micro Systems) and with the following criteria: - ball module - Test mode: compression - Pre-test speed: 1 mm / sec - Test speed: 0.5 mm / sec - Post-test speed: 2 mm / sec - Target mode: distance - Distance: 15 mm - Trigger type: Auto - Trigger force: 2 N

[0284] The force expressed in g is measured using the texturometer.

[0285] The emulsions and the results obtained for the different tests are shown in the following Table:

[0286] [Tables 13] Emulsion with Inv.l Emulsion with Inv.2 Emulsion with CP.l Strength (Pasteurization) 94 g 80 g 5 g Strength (Sterilization) 77 g 72 g 6 g

[0287] The faba bean proteins of the invention make it possible to form emulsions with a firmer texture than that obtained with the comparative protein, whether this emulsion is sterilized or pasteurized. This makes it possible to predict that it can be advantageously used for the manufacture of vegetarian or non-vegetarian emulsified products, such as Strasbourg or Frankfurt sausage substitutes.

Claims

Claims

1. A method for manufacturing a denatured fava bean protein extract comprising the following steps: a) preparing an aqueous suspension of ground fava beans; b) extracting a protein fraction by solid-liquid separation of the aqueous suspension; c) adjusting said protein fraction to a pH of between 4.0 and 5.7; d) heat treating the protein fraction at a temperature of between 50 and 80°C for a time of between 1 and 30 seconds to form a suspension of precipitated proteins; e) solid-liquid separation of the suspension of precipitated proteins to recover a fava bean protein extract having a protein content by weight relative to its dry matter of greater than 70%; f) adjusting the pH to between 6.3 and 8; g) heat treatment of the fava bean protein extract at pH adjusted to a temperature ranging from 85 to 160°C for a period of time sufficient to obtain denaturation of the fava bean protein;h) possibly drying.;

2. Method according to claim 1 characterized in that it comprises, following the heat treatment step g), a step of vacuum cooling of the suspension of precipitated proteins.

3. Method according to one of claims 1 or 2 characterized in that the heat treatment step g) is carried out from 100 to 150°C for 0.1 to 60 seconds, preferably from 0.1 to 10 seconds.

4. Method according to any one of claims 1 to 3 characterized in that it comprises, after step g), a step of shearing the denatured protein extract, for example by passing it through a high pressure or homogenization pump.

5. Process according to any one of claims 1 to 4, characterized in that a starch-rich fraction and / or a fiber-rich fraction is also recovered from the aqueous suspension of crushed field beans.

6. Method according to any one of claims 1 to 5 characterized in that the pH is adjusted between 6.5 and 7.5 during step f).

7. Denatured faba bean protein extract obtainable by the process according to any one of claims 1 to 6.

8. Denatured fava bean protein composition having a fava bean protein content greater than 70%, % expressed by weight relative to the dry matter of the composition, said composition having at least one of the following properties: - solubility at pH 6 greater than or equal to 50%, - solubility at pH 7 greater than or equal to 75%, - emulsion capacity greater than 400 mL / g.

9. Composition according to claim 8, characterized in that the composition has a solubility at pH 6 greater than or equal to 70% or a solubility at pH 7 greater than or equal to 80%.

10. Composition according to any one of claims 8 or 9, characterized in that the composition has an emulsion capacity greater than 500 mL / g, preferably greater than 600 mL / g.

11. Field bean composition according to any one of claims 8 to 10, characterized in that the composition has a gelling power greater than or equal to 100 Pa, for example ranging from 100 to 1000 Pa, preferably ranging from 300 to 800 Pa.

12. Fava bean protein composition according to any one of claims 8 to 11, characterized in that the composition has a fava bean protein content greater than 80%, preferably greater than 90%, % expressed by weight relative to the dry matter of the composition.

13. Fava bean protein composition according to any one of claims 8 to 12, characterized in that the composition has a dry matter greater than 90%, preferably greater than 94%, % expressed by weight relative to the total weight of the composition.

14. A fava bean protein composition according to any one of claims 8 to 13, characterized in that the composition has a denaturation enthalpy of less than 0.5 J / g.

15. A textured faba bean protein composition obtained by texturizing the faba bean protein extract of claim 7 or the faba bean protein composition according to any one of claims 8 to 14.

16. Use of the faba bean protein extract according to claim 7, the faba bean protein composition according to any one of claims 8 to 14, or the textured faba bean protein composition according to claim 15, for the manufacture of food or beverage products.

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