Temperature gelling pea proteins

Abstract

The present invention relates to a functionalized pea protein having, according to test A, a gelling capacity of at least 300 Pa, preferably in the range of 500 Pa to 2000 Pa, most preferably in the range of 600 Pa to 1800 Pa, and a measured viscosity of less than 0.65 Pa.s, which viscosity is measured at 15% by weight of dry matter, a shear rate of 40 s-1, and a temperature of 20 DEG C; a method for obtaining such a protein; and the use of such proteins in food or beverages.

Description

Technical Field

[0001] This invention relates to a temperature-gelled pea protein with further reduced viscosity. A second aspect of the invention relates to a method for manufacturing pea protein. A third aspect relates to the use of said pea protein in food applications. Background Technology

[0002] The daily requirement for protein typically accounts for 12% to 20% of food intake. This protein is provided equally by animal-derived products (meat, fish, eggs, dairy products) and plant-based foods (grains, legumes, seaweed).

[0003] In developed countries, protein intake still primarily comes from animal sources. These proteins possess excellent nutritional properties and interesting functional properties, making them suitable for use in a wide variety of foods.

[0004] However, numerous studies have shown that excessive consumption of animal-derived protein, which impairs plant protein intake, is one of the causes of increased cancer and cardiovascular disease. Furthermore, animal protein has many drawbacks, particularly in terms of its allergenicity (especially proteins from milk or eggs) and its environmental impact related to the harmful effects of intensive farming.

[0005] Therefore, manufacturers are increasingly demanding plant-based proteins that have beneficial nutritional and functional properties but do not have the disadvantages of animal-derived proteins.

[0006] Since the 1970s, peas have been the most widely developed legume in Europe (primarily France), especially as a protein source intended for use in animal and human food. Peas contain approximately 27% by weight of protein. The term "pea" is considered herein to be used for its most widely accepted purpose and specifically includes all wild varieties of "smooth pea" and all mutant varieties of "smooth pea" and "wrinkled pea," regardless of the intended use of said varieties (human food, animal feed, and / or other uses). Pea protein (primarily pea globulin) has been extracted and utilized industrially for many years. As an example of a method for extracting pea protein, patent EP1400537 can be cited. In this method, the seeds are ground in the absence of water (a method known as "dry grinding") to obtain a powder. This powder is then suspended in water at room temperature, and the various steps for protein extraction are then performed.

[0007] Despite the undeniable properties of pea-derived proteins, they exhibit a lower temperature gelling ability than other plant proteins. For example, soy protein isolates possess excellent temperature gelling capabilities. However, for many finished products (proteins structured via dry or wet extrusion, meat substitutes such as emulsified sausages, deli substitutes, egg substitute compositions, or creamy desserts), the use of temperature-gelled proteins allows for the provision of foods with improved textures.

[0008] Because it is generally believed that preserving the natural structure of a protein allows it to retain its function, methods for producing proteins with improved temperature-gelling capabilities typically involve the following steps: after these steps, the protein's natural characteristics are maintained. Indeed, this is beneficial for the formation of gelling ability when used in the final food product. Document WO2022 / 174339 describes a method for processing legume proteins (particularly undenatured pea protein concentrates and isolates) to induce gelation under heating and in a brine solution. This method includes maintaining the protein at an alkaline or acidic pH prior to neutralization and / or cold atmospheric plasma treatment. Document WO2021 / 260169 also describes a protein with improved temperature-gelling capabilities. This protein is obtained through a method involving acid washing and alkali washing.

[0009] However, other methods for providing proteins with improved temperature-gelling ability have been described, during which the protein denatures. For example, WO2023 / 076541 describes the use of transglutaminase to form a gel in an aqueous protein composition. However, this document does not describe how to provide proteins with improved gelling ability and reduced viscosity: the method modifies the protein structure by cross-linking the protein (and cross-linking increases the protein's viscosity), and cannot provide proteins with improved gelling ability in a dry form, let alone in the form of ready-to-use protein powder. Furthermore, the use of transglutaminase results in a significant alteration of the protein's primary structure. Document WO 2020 / 221978 describes the manufacture of pea protein powder with improved gelling ability, in which a micronization step is performed. However, the gelling ability may still be insufficient, and this document does not mention the viscosity of the resulting product. However, the inventors have observed that micronized protein powder has a higher viscosity than non-micronized powder. Document WO2011 / 124862 describes a method for manufacturing soluble functional pea protein, which includes a heat treatment step at a temperature of 100°C to 160°C for 0.01 s to 1 s, followed by a cooling step. As demonstrated below in the Examples section of this application, although heat treatment can provide the protein with temperature gelling ability, it is still insufficient.

[0010] Regarding isoelectrically precipitated plant protein concentrates, EP522800 A2 describes a method for producing proteins with improved temperature-gelling ability. This method includes a step of suspending the protein at an alkaline pH for 1-120 minutes at a temperature of 75°C-95°C prior to neutralization and spray drying. One problem with this method is that it causes a significant increase in product viscosity (the viscosity of a 17% dry matter suspension of the product treated in this manner is five times that of the suspension of the product before treatment). Therefore, while methods exist to improve the gelling ability of certain proteins, this improvement is accompanied by significant changes in other properties, particularly a significant increase in viscosity. In the Examples section, commercial pea proteins with improved temperature-gelling ability are also referred to: all of these pea proteins also have a much higher viscosity than pea proteins with reduced gelling ability.

[0011] In summary, we need proteins that, in addition to their temperature-gelling ability, also have limited viscosity.

[0012] Through extensive research, the applicant has developed a novel manufacturing method that can provide pea proteins with high gelling ability but reduced viscosity. Therefore, unlike previously described pea proteins, these proteins can be used to provide fluid-liquid compositions, even at high concentrations. These proteins are also capable of forming gels upon heating, which is highly advantageous for manufacturing the aforementioned finished products. Summary of the Invention

[0013] Therefore, the object of the present invention is a functionalized pea protein that, according to test A, has a gelling ability of at least 300 Pa, preferably in the range of 500 Pa to 2000 Pa, and most preferably in the range of 600 Pa to 1800 Pa, and a measured viscosity of less than 0.65 Pa·s, which is measured at 15% by weight of dry matter, a shear rate of 40 s⁻¹, and a temperature of 20°C.

[0014] Another object of the present invention is a method for manufacturing the functionalized pea protein of the present invention, the method comprising:

[0015] - Provides an aqueous pea protein composition comprising pea peptides and pea protein precipitated at isoelectric pH, wherein the peptides are insoluble at pH below 8.5, and the pea protein in the aqueous composition has a denaturation percentage of less than 65%.

[0016] - Adjust the pH of the aqueous composition to the range of 6.0 to 6.8.

[0017] - The mixture is heat-treated to obtain functionalized pea protein.

[0018] Therefore, the protein manufacturing method developed by the inventors enables the production of functionalized pea proteins with reduced viscosity, combining temperature-gelling capabilities with properties distinct from known gelling pea proteins. To obtain such functionalized pea proteins, the inventors used a method involving heat treatment of pea proteins pre-placed within a restricted pH range of 6.0 to 6.8. As shown in the Examples section, within this restricted range, the functionalized pea proteins exhibit temperature-gelling capabilities while maintaining a target viscosity.

[0019] For providing a heat-treated aqueous pea protein composition, a method for extracting insoluble peptides (i.e., proteins not present when flour is extracted in aqueous solution at its natural pH below 8.5) is used. This extraction can be performed, for example, by placing pea material (pea flour, pea fiber, or insoluble components obtained from peas, etc.) in an alkaline solution. These peptides can be recovered at different stages of the method as detailed below. The aqueous pea protein composition also contains proteins precipitated at isoelectric pH. All steps involved in providing the aqueous pea protein composition are performed conservatively to at least partially preserve the natural characteristics of the heat-treated protein; this is a key criterion for obtaining the functionalized pea protein of the present invention, as shown in the Examples section.

[0020] Another object of the present invention relates to the use of functionalized pea protein according to the invention in the manufacture of food or beverages, particularly proteins textured by dry or wet extrusion, meat substitutes such as emulsified sausages, deli substitutes, egg substitute compositions, or cream desserts.

[0021] The present invention will now be described in detail. Attached Figure Description

[0022] Other features, details, and advantages will become apparent from reading the following detailed description and analyzing the accompanying drawings, in which:

[0023] Figure 1

[0024] [ Figure 1 The figure shows the variation of residual protein content in the insoluble fraction extracted during the aqueous extraction of pea material with extraction pH.

[0025] Figure 2

[0026] [ Figure 2 [This shows a wet extruded strip Inv.1 obtained from a mixture of pea protein, pea fiber and potato starch obtained in Example 1 using an extrusion profile 1.]

[0027] Figure 3

[0028] [ Figure 3 The wet extrusion strip CP.1 is shown, obtained from a mixture containing pea protein of Counterexample 5 obtained by extrusion profile 1.

[0029] Figure 4

[0030] [ Figure 4 [This shows a wet extruded strip Inv.2 obtained from a mixture containing pea protein of Example 1 obtained using extrusion profile 2.]

[0031] Figure 5

[0032] [ Figure 5 The wet extrusion strip CP.2 is shown, obtained from a mixture containing pea protein of Counterexample 5 obtained using extrusion profile 2.

[0033] Figure 6

[0034] [ Figure 6 The image shows how to scoop out pea protein from commercially available NUTRALYS using a spoon. ® The F85 M mixture yields a texture comparable to cream.

[0035] Figure 7

[0036] [ Figure 7 The image shows the texture of cream obtained when scooped with a spoon from a mixture containing pea protein of the present invention.

[0037] Figure 8

[0038] [ Figure 8 The image shows the texture of commercially available butter when scooped with a spoon. Detailed Implementation

[0039] This invention relates to a functionalized pea protein that exhibits temperature gelling ability and reduced viscosity.

[0040] For the purposes of this invention, pea protein is an extract derived from peas that primarily contains protein and a smaller proportion of other components, as described later in the specification.

[0041] "Functionalized pea protein" refers to pea protein used in methods for obtaining the gelling ability and viscosity of the protein provided by the present invention.

[0042] More specifically, the "temperature gelling ability" is characterized by a gelling capacity of at least 300 Pa as determined by test A.

[0043] Gelation ability: Test A

[0044] The term "gelling ability" refers to a functional property consisting of the ability of a protein composition to form a gel or network, which increases viscosity and produces a state of matter between a liquid and a solid. The term "gel strength" may also be used. To quantify this gelling ability, it is necessary to generate the network and evaluate its strength. For this quantification, Test A is used in this invention, described as follows:

[0045] 1) Dissolve the tested protein composition in water with 15% + / - 2% solids and pH 7 at 60℃ ± 2℃;

[0046] 2) Stir at 60℃±2℃ for 5 minutes;

[0047] 3) Cool to 20℃±2℃ and stir at 350rpm for 24 hours;

[0048] 4) Suspension was achieved using a controlled stress rheometer equipped with concentric cylinders;

[0049] 5) Measure the elastic modulus G' and viscous modulus G'' by applying the following temperature characteristics:

[0050] a. Stage 1: After stabilizing at 20℃±2℃ and heating from 20℃±2℃ to 80℃±2℃ within 10 minutes, measure parameter G'1;

[0051] b. Stage 2: Stabilize at 80℃±2℃ for 110 minutes; c. Stage 3: Cool from 80℃±2℃ to 20℃±2℃ within 30 minutes, and measure G'2 after stabilizing at 20℃±2℃;

[0052] 6) Calculate the gelling capacity equal to G'2-G'1.

[0053] In a preferred embodiment, the controlled stress rheometer is selected from models DHR 2 (TA, Instruments) and MCR 301 (Anton Paar), featuring concentric cylindrical spindles. These are equipped with a temperature control system based on the Peltier effect. To avoid evaporation problems at high temperatures, liquid paraffin is added to the top of the sample.

[0054] For the purposes of this invention, a "rheometer" is a laboratory instrument used to measure the rheological properties of fluids or gels. It applies force to a sample. Typically, due to its characteristically small size (very small mechanical inertia of the rotor), it allows for fundamental studies of the mechanical properties of liquids, gels, suspensions, pastes, etc., in response to the applied force.

[0055] The so-called "controlled stress" model allows for the determination of the intrinsic viscoelasticity of a material by applying sinusoidal stress (oscillatory mode), which is particularly dependent on time (or angular velocity ω) and temperature. In particular, this type of rheometer allows for the acquisition of the complex modulus G*, which in turn allows for the acquisition of the modulus G', or elastic portion, and G'', or viscous portion.

[0056] The first three steps consist of the following processes: resuspending the protein in water, using precise conditions to maximize subsequent measurements.

[0057] The water selected is preferably reverse osmosis water.

[0058] The temperature is 60°C ± 2°C during the initial resuspension (steps 1 and 2), and then 20°C ± 2°C after 24 hours of dissolution and cooling before measurement (step 3). Generally, and unless otherwise specified, when temperatures are given in this specification, they always include a variation of ± 2°C, such as 20°C ± 2°C or 80°C ± 2°C.

[0059] A defined amount of protein is added to the water to obtain a suspension containing 15% ± 2% solids. To do this, equipment well known to those skilled in the art, such as beakers and stir bar, is used. A volume of 50 ml is stirred at 350 rpm at room temperature for at least 10 hours. Generally, and unless otherwise specified, the solids content given in this specification always includes a variation of ± 2%, for example, 15% ± 2%. The pH is adjusted to 7 ± 0.5 (20°C) using a pH meter and acid-base reagents, as is well known in the art.

[0060] The fourth step consists of the following process: introducing the sample into the rheometer and covering the sample with a thin oil layer to limit evaporation.

[0061] During the fifth step, the following temperature scheme is then applied: a. Stage 1: heating from 20℃±2℃ to 80℃±2℃ within 10 minutes; b. Stage 2: stabilizing at 80℃±2℃ for 110 minutes; c. Stage 3: cooling from 80℃±2℃ to 20℃±2℃ within 30 minutes.

[0062] During this process, parameter G' is continuously measured and recorded.

[0063] The sixth and final steps of test A involve utilizing this record. Two values ​​are extracted: G'1 = the G' value at the start of stage 1 after stabilization at 20℃±2℃, and G'2 = the G' value at the end of stage 3 after stabilization at 20℃±2℃.

[0064] The gelling capacity is equal to G'2 - G'1.

[0065] Preferably, the gelling capacity determined according to test A is in the range of 500 Pa to 2000 Pa, most preferably 600 Pa to 1800 Pa, or 600 Pa to 1000 Pa, or 1000 Pa to 2000 Pa, or 1100 Pa to 1800 Pa. The gelling capacity can be greater than or equal to 400 Pa, greater than or equal to 500 Pa, greater than or equal to 600 Pa, greater than or equal to 700 Pa, greater than or equal to 800 Pa, greater than or equal to 900 Pa, greater than or equal to 1000 Pa, or greater than or equal to 1100 Pa. The gelling capacity can be less than or equal to 2000 Pa, less than or equal to 1900 Pa, less than or equal to 1800 Pa, less than or equal to 1700 Pa, less than or equal to 1600 Pa, less than or equal to 1500 Pa, less than or equal to 1400 Pa, less than or equal to 1300 Pa, less than or equal to 1200 Pa, less than or equal to 1100 Pa, or less than or equal to 1000 Pa.

[0066] According to the present invention, the reduced viscosity of functionalized pea protein can be expressed as a viscosity of less than 0.65 Pa·s (15% dry matter, 40 s⁻¹, 20 °C).

[0067] According to the present invention, viscosity can be determined using test B.

[0068] Viscosity: Test B

[0069] To determine the viscosity characteristics in water, a pea protein aqueous solution containing 15% dry matter (reverse osmosis water azide-treated at 200 ppm to prevent any bacteriological risk) was measured in an AR2000 rheometer from TA Instruments, which has a concentric cylindrical geometry and a shear rate from 0.6 × 10⁻⁶ over 3 minutes. -3 up to 600s -1 (Logarithmic) and the temperature is 20°C (temperature equilibration for 3 minutes before testing). Stir the solution at 750 rpm and 20°C for at least 10 hours before measurement. Do not adjust the pH. Report the viscosity at 40 s⁻¹ as the viscosity according to test B.

[0070] According to test B, the viscosity of functionalized pea protein can be less than or equal to 0.60 Pa·s, for example less than or equal to 0.55 Pa·s, particularly less than 0.5 Pa·s, advantageously less than 0.4 Pa·s, preferably 0.05 Pa·s to 0.4 Pa·s, for example 0.1 Pa·s to 0.4 Pa·s, or 0.05 Pa·s to 0.65 Pa·s, for example 0.1 Pa·s to 0.60 Pa·s.

[0071] Preferably, the functionalized pea protein of the present invention has a solubility of less than or equal to 49%, preferably less than or equal to 45%, for example, in the range of 20% to 40%, according to test C.

[0072] Solubility: Test C

[0073] Solubility was determined according to the following test C method:

[0074] Measuring solubility in water

[0075] The measurement is based on diluting the sample in distilled water, centrifuging, and analyzing the supernatant.

[0076] program :

[0077] 150g of distilled water was introduced into a 400ml beaker at 20℃±2℃, mixed with a magnetic stir bar, and 5g of the sample to be tested was added precisely.

[0078] Adjust the pH to the desired value using 0.1N NaOH or HCl (pH 7), or leave the pH unchanged.

[0079] The total water content is 200g.

[0080] Mix at 1000 rpm for 30 minutes, then centrifuge at 3000 g for 15 minutes.

[0081] Collect 25g of supernatant.

[0082] It is introduced into a pre-dried and weighed crystallizer.

[0083] Place in an oven at 103℃±2℃ for 1 hour.

[0084] Then place it in a desiccator (with desiccant) to cool to ambient temperature and weigh it.

[0085] The soluble solids content, expressed as a percentage by weight, is given by the following formula:

[0086] [Formula 1]

[0087]

[0088] in:

[0089] -P = Sample weight (in grams) = 5g

[0090] -m1 = Weight of the crystallizer after drying (in grams)

[0091] -m2 = weight of the empty crystallizer (in grams).

[0092] -P1 = Weight of collected sample (in grams) = 25g

[0093] Functionalized pea proteins can have a pH range of 6.0 to 6.8.

[0094] According to test C, the solubility of functionalized pea protein at pH 7 can be less than or equal to 49%, preferably less than or equal to 45%, and more preferably 20% to 40%.

[0095] The functionalized pea protein of the present invention can have a protein content ranging from 80% to 90%.

[0096] Functionalized pea protein is typically a protein composition that contains a few other components besides protein, such as starch, lipids, fiber, and / or sugars.

[0097] The protein content is the N6.25 content calculated using the Dumas method. Typically, the total starch content in pea protein produced according to the method of the present invention ranges from 0% to 20%, for example, 0% to 10%, particularly 0.5% to 5%. This total starch content can be measured using the AOAC 996.11 method. Generally, the total fiber content ranges from 0% to 20%, for example, 1% to 18%, particularly 2% to 10%. This content can be determined using the AOAC 2017.16 method. Typically, the total lipid content ranges from 0% to 15%, for example, 1% to 10%. The total lipid content can be determined using the AOAC 996.06 acid hydrolysis method. The sugar content ranges from 0% to 10%, typically 0.5% to 5%. The sugar content can be determined by high-performance liquid chromatography (HPLC).

[0098] All the above contents are expressed on a dry matter basis.

[0099] According to one implementation, the functionalized pea protein has a denaturation percentage of greater than 75%, or even greater than 90%, most preferably about 100%.

[0100] The denaturation percentage of pea protein is a quantification relative to the denaturation of pea protein isolated by precipitation at isoelectric pH without heat input, and which was not subjected to any heat treatment during its manufacture to make it as natural as possible. Therefore, to determine the denaturation percentage, the inventors prepared pea protein by extracting pea flour in cold water at pH 9 and then precipitating it at isoelectric pH (pH 5) at room temperature. The precipitated protein was washed once with cold water to ensure its purity and then freeze-dried. Under these preparation conditions, the pea protein precipitated at isoelectric pH was considered to have the least degree of denaturation, and this pea protein isolate was considered a standard.

[0101] The denaturation percentage of the sample is determined using the denaturation enthalpy of the standard according to the following formula:

[0102] [Formula 2]

[0103] % denaturation = (ΔHd standard - ΔHd sample) / ΔHd standard × 100

[0104] Depending on the method used to determine the denaturation enthalpy (ΔHd), the ΔHd of the sample and the ΔHd of the standard can vary. Under the test conditions D below, the denaturation enthalpy of this standard measured for the pea protein suspension is 2.4 J / g suspension; that is, using the protein content N6.25 based on its dry matter content and the dry matter content of the suspension, the enthalpy of the standard protein ΔHd standard is 12.5 J / g protein (N6.25). However, another method can be used to determine the denaturation enthalpy.

[0105] Enthalpy of denaturation: Test D

[0106] In test D, pea protein was dissolved in water at 20% ± 2% dry matter, and the pH was adjusted to 6.2 by adding 0.1N sodium hydroxide or 0.1N hydrochloric acid if necessary. The solution was stirred at 350 rpm for 2 hours at room temperature. The enthalpy ΔH of the protein was determined by digital subtraction calorimetry (DSC) according to known methods and expressed as N 6.25 per g of protein in the sample. 10-15 mg of this solution sample was placed in a sealed crucible, and the enthalpy was determined by calorimetry. The analysis was performed by heating a suspension of pea protein extract containing 20% ​​dry matter from 5°C to 120°C at 10°C / min. The enthalpy of the protein, expressed as J / g protein, was obtained using the enthalpy determined on a dry matter basis, N 6.25, and the dry matter content.

[0107] DSC calorimeters available for measurement include, for example, the Q20 (TA Instruments), DSC 8000 (PerkinElmer), and DSC (Mettler). Analysis is performed from 5°C to 120°C at a heating rate of 10°C / min.

[0108] According to another embodiment, the residual denaturation enthalpy of the functionalized pea protein is less than 0.5 J / g protein N6.25, preferably less than 0.2 J / g protein N6.25, or even zero, as determined by TEST D.

[0109] Advantageously, the functionalized pea protein of the present invention has a dry matter content of greater than 90%, and most preferably greater than 94%, relative to the weight of the functionalized pea protein on a dry matter basis. The functionalized pea protein can be in powder form, and its particle size d50 can vary considerably, for example from 10 µm to 500 µm, for example from 50 µm to 500 µm, typically from 50 µm to 150 µm. Particle size can be readily measured by laser diffraction.

[0110] To extract the pea protein of the present invention, the inventors have developed a mild extraction method comprising heat-treating an aqueous pea protein composition at a given pH. More specifically, a method for manufacturing the functionalized pea protein of the present invention comprises:

[0111] - Provides an aqueous pea protein composition comprising pea peptides and pea protein precipitated at isoelectric pH, wherein the peptides are insoluble at pH below 8.5, and the pea protein in the aqueous composition has a denaturation percentage of less than 65%.

[0112] - Adjust the pH of the aqueous composition to the range of 6.0 to 6.8.

[0113] - A step of heat-treating the mixture to obtain functionalized pea protein.

[0114] The aqueous pea protein composition is in the form of a suspension mainly comprising water and pea protein. Those skilled in the art can readily adjust the dry matter content of the aqueous composition according to the equipment used, and its range can be from 5% to 25%, for example, from 10% to 18%.

[0115] According to the present invention, the pea protein in the provided aqueous composition is extracted in a manner that does not significantly denature it. As will become apparent in the remainder of the specification, the protein can be denatured in multiple method steps, and based on the description given below, those skilled in the art will be able to select the conditions of the method of the present invention to conserve protein during its extraction, thereby providing an aqueous pea protein composition that can be used in the heat treatment step. Generally, it is preferable to perform the various steps of the method one after another to maintain the undenatured characteristics of the pea protein in the aqueous composition prior to the heat treatment step. This is even more important in steps involving heat. Preferably, to maintain the desired high denaturation percentage, the method uses cold water in the temperature range of 1°C to 25°C, warm water (60°C), or water in the temperature range of 25°C to 60°C.

[0116] Therefore, as mentioned above, the pea protein in the aqueous composition can have a denaturation percentage of less than 65%. Advantageously, the pea protein contained in the aqueous composition has the following denaturation percentage:

[0117] - Less than 60%, or even less than 55%, or even less than 50% and / or

[0118] - Greater than 10%, or even greater than 20%, or even greater than 30%.

[0119] To determine the percentage of denaturation, the protein thus formed during this method can be recovered and freeze-dried in a manner that does not denature it, and then its enthalpy can be measured as above.

[0120] The pea protein in the aqueous composition comprises pea protein precipitated at isoelectric pH.

[0121] The isoelectric point (pH) is the pH at which the net charge of the proteins in a protein fraction is close to zero. This pH can be adjusted to, for example, between 4.0 and 5.7 or between 4.8 and 5.2. When a composition containing pea protein is placed at this pH, some proteins, particularly at least some globulins, will tend to precipitate. Therefore, pea protein precipitated at the isoelectric pH is pea protein that can be separated by solid-liquid separation. Except for the methods described in the tests, pH values ​​are determined at 10% dry matter and 20°C throughout this document.

[0122] The pea protein in the aqueous compositions also contains pea peptides that are insoluble at pH values ​​below 8.5. As will become apparent in the remainder of the specification, and particularly in the different presentations of the pea protein in the aqueous compositions, the pea peptides may or may not precipitate at isoelectric pH. In the variants where they do not precipitate at isoelectric pH, the natural characteristics of the pea protein provided prior to the heat treatment step are enhanced, as the isoelectric precipitation step of the protein can lead to a degree of denaturation.

[0123] "Pea polypeptides that are insoluble at pH below 8.5" should be understood to mean a fraction of pea protein (soluble fraction) extracted in solution by placing pea material in an aqueous composition with a pH greater than 8.5, for example at a mass concentration of 20% relative to the total weight of the aqueous composition, and which is not extracted at lower pH.

[0124] When pea material is placed in an aqueous composition to extract protein, the protein dissolves and ends up in the aqueous fraction, while undissolved protein remains in the insoluble fraction along with other insoluble substances such as fiber and starch. However, as... Figure 1As shown, when extracting pea material, the residual protein content in the insoluble fraction decreases with increasing pH: this is because proteins insoluble at a specific pH are soluble at higher extraction pH levels. According to the invention, it is important that the aqueous composition subjected to heat treatment after pH adjustment contains pea proteins that dissolve during the alkaline suspension of pea material in water at a pH greater than 8.5; therefore, these proteins are referred to in the context of the invention as “pea polypeptides insoluble at pH below 8.5”. Various embodiments for providing aqueous compositions of pea proteins suitable for use in the methods of the invention are described below. Their common feature is that they involve alkaline suspensions of pea material at pH greater than 8.5. These embodiments should not be considered as limiting the scope of the invention.

[0125] The pea material from which pea peptides can be extracted can be any material containing pea peptides that are insoluble at a pH below 8.5. Specifically, the pea material can be pea flour, pea protein concentrate, pea fiber, or an insoluble fraction obtained from peas.

[0126] According to one embodiment, pea flour is obtained by dry milling. Extraction can be carried out by suspending the pea flour thus obtained in water. "Suspending pea flour in water" should also be understood to mean a suspension obtained by wet milling an aqueous suspension of whole peas or cotyledons, advantageously pre-shelled. According to this embodiment, a suspension of pea flour in water can thus be obtained directly. An example of wet milling is described in application WO2019 / 053387.

[0127] Other pea fractions can be used, such as pea fiber, pea concentrate obtained by mechanical dry fractionation of pea flour, or other insoluble fractions obtained from peas. For the extraction of pea peptides that are insoluble at pH below 8.5, it is obviously preferred that these pea fractions have not undergone prior protein extraction in an alkaline suspension at a pH greater than 8.5.

[0128] Different ways of providing aqueous pea protein compositions will now be described.

[0129] According to the first embodiment, the aqueous pea protein composition provided includes:

[0130] - Suspend pea flour in water

[0131] - Separate suspensions to provide primary aqueous solutions and insoluble fractions.

[0132] - Suspend at least a portion of the insoluble fraction in water with a pH greater than 8.5 to form an alkaline suspension.

[0133] - Separate the alkaline suspension to provide a residual solids fraction and a secondary aqueous solution containing dissolved pea peptides.

[0134] - Mix the primary and secondary aqueous solutions to form a mixture of primary and secondary aqueous solutions.

[0135] - The mixture of primary and secondary aqueous solutions is adjusted to an isoelectric pH to form an aqueous pea protein suspension containing pea protein and pea peptides precipitated at the isoelectric pH.

[0136] - Isolate pea protein from an aqueous pea protein suspension to form an aqueous pea protein composition.

[0137] According to the second embodiment, the aqueous pea protein composition includes:

[0138] - Suspend pea flour in water

[0139] - Separate suspensions to provide primary aqueous solutions and insoluble fractions.

[0140] - Adjust the primary aqueous solution to an isoelectric pH to form an aqueous suspension of precipitated pea protein.

[0141] - Separate the precipitated pea protein from an aqueous suspension of precipitated pea protein to form an aqueous composition of precipitated pea protein.

[0142] - Suspend at least a portion of the insoluble fraction in water with a pH greater than 8.5 to form an alkaline suspension.

[0143] - Separate the alkaline suspension to provide a residual solids fraction and a secondary aqueous solution containing pea peptides.

[0144] - The aqueous composition of precipitated pea protein is mixed with a secondary aqueous solution to form an aqueous pea protein composition.

[0145] The first and second implementation schemes have the advantage of maximizing the recovery of pea peptides while limiting the amount of alkali required for extraction.

[0146] According to the third embodiment, the aqueous pea protein composition includes:

[0147] -The pea flour is suspended in alkaline water with a pH greater than 8.5 to form an alkaline pea flour suspension.

[0148] - Separate an alkaline pea flour suspension to provide an aqueous solution containing pea protein and an insoluble fraction, the pea protein comprising pea peptides.

[0149] - Adjust the aqueous solution to an isoelectric pH to form an aqueous pea protein suspension containing precipitated pea protein and pea peptides.

[0150] - Separate the precipitated pea protein from the suspension to form an aqueous pea protein composition.

[0151] The third implementation scheme has the advantage of involving fewer individual steps than other implementation schemes.

[0152] According to the fourth embodiment, the aqueous pea protein composition includes:

[0153] - Suspend pea flour in water

[0154] - Separate the suspension to provide insoluble fractions and a primary aqueous solution containing residues of suspended pea peptides.

[0155] - Separate the primary aqueous solution to provide a purified primary aqueous solution and residues containing pea peptides.

[0156] - Suspend at least some of the residues containing pea peptides in water with a pH greater than 8.5 to form an alkaline suspension.

[0157] - Separate the alkaline suspension to provide residual fractions and secondary aqueous solutions containing pea peptides.

[0158] - The purified primary aqueous solution and secondary aqueous solution are mixed to form a mixture of aqueous solutions.

[0159] - Adjust the mixture of aqueous solutions to an isoelectric pH to form an aqueous pea protein suspension.

[0160] - Isolate pea protein from a suspension to form an aqueous pea protein composition.

[0161] According to the fifth embodiment, the aqueous pea protein composition includes:

[0162] - Suspend pea flour in water

[0163] - Separate the suspension to provide insoluble fractions and a primary aqueous solution containing residues of suspended pea peptides.

[0164] - Separate the primary aqueous solution to provide a purified primary aqueous solution and residues containing pea peptides.

[0165] - Suspend at least some of the residues containing pea peptides in water with a pH greater than 8.5 to form an alkaline suspension.

[0166] - Separate the alkaline suspension to provide residual fractions and secondary aqueous solutions containing pea peptides.

[0167] - Adjust the primary aqueous solution to an isoelectric pH to form a suspension of precipitated pea protein.

[0168] - Separate the precipitated pea protein from the suspension of precipitated pea protein.

[0169] - The precipitated pea protein is mixed with a secondary aqueous solution to form an aqueous pea protein composition.

[0170] According to the sixth embodiment, the aqueous pea protein composition provided includes:

[0171] - Suspend pea flour in water

[0172] - Separate the suspension to provide an aqueous solution containing a residue and insoluble fractions of suspended pea peptides.

[0173] - At a pH greater than 8.5, an aqueous solution containing the residue of suspended pea peptides is suspended to form an alkaline suspension.

[0174] - Separate alkaline suspensions to provide aqueous solutions and residual solids fractions.

[0175] - Adjust the aqueous solution to an isoelectric pH to form an aqueous pea protein suspension.

[0176] - Isolate pea protein from a suspension to form an aqueous pea protein composition.

[0177] Typically, the amount of residues containing pea peptides is in the range of 5% to 20% of the dry matter of the aqueous solution containing them.

[0178] According to the seventh embodiment, the aqueous pea protein composition includes:

[0179] - Suspend pea flour in water to form a pea flour suspension.

[0180] - Separate pea flour suspensions to provide primary aqueous solutions and insoluble fractions.

[0181] - Separate insoluble fractions to form starch and fiber fractions.

[0182] - Suspend at least a portion of the fiber fraction in water with a pH greater than 8.5 to form an alkaline suspension.

[0183] - Separate the alkaline suspension to provide a residual solids fraction and a secondary aqueous solution containing dissolved pea peptides.

[0184] - To mix primary and secondary aqueous solutions to form a mixture of aqueous solutions.

[0185] - The mixture of aqueous solutions is adjusted to an isoelectric pH to form an aqueous pea protein suspension containing pea protein and pea peptides precipitated at the isoelectric pH.

[0186] - Isolate pea protein from a suspension to form an aqueous pea protein composition.

[0187] According to the eighth embodiment, the aqueous pea protein composition includes:

[0188] - Suspend pea flour in water to form a pea flour suspension.

[0189] - Separate pea flour suspensions to provide primary aqueous solutions and insoluble fractions.

[0190] - Separate insoluble fractions to form starch and fiber fractions.

[0191] - Adjust the primary aqueous solution to an isoelectric pH to form an aqueous suspension of precipitated pea protein.

[0192] - Separate precipitated pea protein from a suspension to form an aqueous composition of precipitated pea protein.

[0193] - Suspend at least a portion of the fiber fraction in water with a pH greater than 8.5 to form an alkaline suspension.

[0194] - Separate the alkaline suspension to provide a residual solids fraction and a secondary aqueous solution containing pea peptides.

[0195] - The aqueous composition of precipitated pea protein is mixed with a secondary aqueous solution to form an aqueous pea protein composition.

[0196] Compared to other embodiments, the fourth, fifth, sixth, seventh, and eighth variants have the advantage of using a smaller amount of alkali because a smaller amount of pea material is suspended in an alkaline state compared to flour (third embodiment) or the entire insoluble fraction from the first separation (first and second embodiments). By selecting and adjusting the separation apparatus, i.e., by performing separation in a manner that keeps the residue in suspension, an aqueous solution containing suspended pea peptides is obtained, which contain pea peptides that can be used in this invention.

[0197] The separation step can be carried out, in particular, by at least one separation step using a decanter (especially a decanter centrifuge, centrifuge) or a separation system that uses centrifugal force to separate components by utilizing a centrifugal force field caused by the movement of the mixture in a stationary device (such as a hydrocyclone). Those skilled in the art will be able to select the most suitable equipment and operating conditions to obtain the different fractions described above.

[0198] Preferably, the aqueous pea protein composition further comprises pea peptides insoluble at pH values ​​below 8.7 or 8.8. Preferably, the aqueous pea protein composition further comprises pea peptides insoluble at pH values ​​below 9.0. Preferably, the aqueous pea protein composition further comprises pea peptides insoluble at pH values ​​below 9.2. It is readily understood that these insoluble pea peptides (similar to pea peptides insoluble at pH values ​​below 8.5) are obtained through alkaline suspensions at pH values ​​greater than 8.7, greater than 8.8, greater than 9.0, and greater than 9.2, respectively.

[0199] To obtain pea protein precipitated at isoelectric pH, an aqueous solution of dissolved pea protein is typically adjusted to an isoelectric pH to form an aqueous suspension of pea protein precipitated at isoelectric pH, followed by separation of the pea protein from the suspension. As mentioned above, the pH can be set between 4.0 and 5.7, or even between 4.8 and 5.2. The pH can be adjusted by adding an organic or inorganic acid, such as hydrochloric acid, sulfuric acid, or citric acid, or a mixture thereof. This step e) can be carried out in a stirred or unstirred tank. It can be longer or shorter, occurring almost instantaneously, or lasting from 1 to 240 minutes, for example, typically 5 to 60 minutes. This addition of base or acid can be done online, and the acid can be in the form of an aqueous solution.

[0200] During this isoelectric precipitation, it is important to limit heat input to maintain the denaturation percentage of the present invention. Therefore, proteins can be precipitated by thermal coagulation, but this must be done in a manner that maintains the denaturation percentage as defined above. It is advantageous to use a pasteurization stage at a temperature below 80°C, advantageously for a short duration, which can be less than 30 seconds. Preferably, the pasteurization stage is carried out at a temperature ranging from 45°C to 77°C, advantageously from 60°C to 72°C. Preferably, the pasteurization stage lasts from 1 to 10 seconds. Heat input via direct steam injection is also preferably limited or even eliminated, as it is likely to denature the proteins. Alternatively, preheating can be performed using a heat exchanger (e.g., a plate heat exchanger) followed by heating via direct steam injection to limit heat input via this direct steam injection. Pasteurization (especially under the preferred conditions described above) has the advantage of rapidly coagulating and precipitating pea protein, thereby accelerating the manufacturing process while preserving the natural characteristics of the protein. Advantageously, during the pasteurization stage, the temperature is rapidly reduced, for example, by a vacuum (known to those skilled in the art as "rapid cooling"), the applied vacuum being determined according to the selected cooling temperature. This cooling also preserves the natural characteristics of pea protein.

[0201] Once provided, the aqueous pea protein composition is pH-adjusted to a range of 6.0 to 8.0, preferably 6.0 to 6.8. This pH adjustment is typically achieved by adding an alkali such as sodium hydroxide, potassium hydroxide, calcium hydroxide, or mixtures thereof. Optionally, the aqueous composition may be diluted such that the pH-adjusted aqueous pea protein composition has a dry matter content that allows it to be heat-treated in selected equipment. Preferably, the pH range is 6.1 to 6.5. Preferably, this pH adjustment step between providing the aqueous pea protein composition and heat treatment is performed as quickly as possible and lasts for less than 60 minutes, so that the pea protein retains its denaturing enthalpy prior to the heat treatment step.

[0202] A heat treatment step is then performed to obtain the functionalized pea protein of the present invention. Heat treatment is typically characterized by a time / temperature scale. Those skilled in the art will know how to adjust the conditions to obtain the functionalized pea protein of the present invention.

[0203] The temperature range during the heat treatment step of the mixture can be from 105°C to 128°C, preferably from 110°C to 125°C, and most preferably 120°C. Preferably, the heat treatment step of the mixture lasts from 0.1 to 20 seconds, advantageously from 0.1 to 5 seconds. The method of the present invention generally includes a cooling step of the functionalized pea protein after the heat treatment step. According to a preferred variant, this cooling step is achieved by rapid cooling. At the end of this step, the temperature can vary depending on the temperature of the previous step. Generally, the cooling temperature is selected such that the difference between the heat treatment temperature and the cooling temperature is in the range of 10°C to 80°C, for example, from 30°C to 70°C. The cooling temperature can be in the range of 30°C to 100°C, and advantageously below 80°C, for example, from 60°C to 80°C, for example, about 70°C. By way of example, in an advantageous embodiment where the heat treatment temperature range is from 105°C to 128°C, the cooling temperature can be in the range of 60°C to 80°C, for example, about 70°C.

[0204] According to one variation of the method, it includes a step of shearing the functionalized pea protein, for example, by passing an aqueous dispersion of the protein through a high-pressure pump. As an example of a high-pressure pump, one could mention high-pressure pumps sold by Silverson, also known as "high-shear mixers," such as those in the UHS series. Preferably, the shearing step is performed using a high-pressure pump. According to another variation, the method includes a step of homogenizing the functionalized pea protein.

[0205] The shearing or homogenization step can be performed before or after the heat treatment and / or pH raising steps. However, if these optional steps are performed, they are usually carried out at low intensity without adversely affecting the function of the pea protein, especially its gelling ability.

[0206] The method according to the invention may further include a step of functionalizing and drying pea protein. Generally, this drying step is performed to achieve the aforementioned dry matter content. For this purpose, any technique well known to those skilled in the art can be used, such as freeze-drying, rapid drying, drum drying, or atomization. The method may also include a grinding or micronization step. Atomization is a preferred technique, particularly multi-effect atomization. This technique can be selected so that the powder has the aforementioned d50 particle size. The method of the invention may further include a step of dry milling the protein.

[0207] The present invention also relates to the use of functionalized pea protein according to the invention in the manufacture of food or beverages.

[0208] Generally, the pea protein of the present invention can be used in foods and beverages that may include an amount up to 100% by weight relative to the total dry weight of the food or beverage product, for example, an amount of pea protein from about 1% to about 80% by weight relative to the total dry weight of the food or beverage product. All intermediate amounts (i.e., 2%, 3%, 4%, ..., 77%, 78%, 79% by weight relative to the total weight of the food or beverage product), and all intermediate ranges based on these amounts, may be used. These food and beverage products may be suitable for vegetarians or vegan populations.

[0209] One use of the protein of this invention relates to its use in beverages. In beverages, the protein content of these products can vary considerably and can also be high-protein beverages. The protein content can range, for example, from 1% to 12% of the total weight of the beverage on a dry weight basis, particularly from 3% to 10% of the total weight of the beverage on a dry weight basis. The beverage can be of any type and includes plant-based products containing milk or milk substitutes, including barista milk and coffee creamer. Plant-based milk substitutes can be made from pea protein according to the invention, along with other optional ingredients such as fats, carbohydrates, and / or emulsion-forming substitutes. Alternatively, milk substitutes can be made from “plant milk” obtained from plants, such as, for example, oat milk, rice milk, soy milk, coconut milk, or almond milk. Thus, these plant milks can supplement the protein of the invention. These may also include other acidic or non-acidic ready-to-drink beverages, 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 water, fruit juice, and sweetened or unsweetened tea or coffee beverages); alcoholic beverages such as beer or spirits, smoothies, and beverage concentrates (including but not limited to liquid concentrates and syrups, and non-liquid "concentrates," such as freeze-dried and / or powdered formulations or "powder mixtures"). It should be noted that flavoring agents or masking agents are commonly used in beverages to reduce the pea flavor characteristics or bitter aftertaste of the protein, or to flavor the beverage. One advantage of the pea protein of this invention is that using it instead of conventional pea protein allows for a reduction in the amount of flavoring agents or masking agents used, or even complete elimination of these components from the beverage, while retaining a very satisfying flavor. The beverage may also include hydrocolloids.

[0210] The food products that may be involved include baked goods, such as bread products (including but not limited to sourdough and unleavened bread, sandwich bread, yeast bread and unleavened bread such as baking soda bread), bread containing all types of wheat flour, bread containing all types of flour other than wheat flour (such as potato flour, rice flour, barley flour, spelt flour and rye flour), gluten-free bread; mixtures for preparing said baked goods; sweet baked goods (including but not limited to bread rolls, cakes, pies, pastries, waffles, pancakes, muffins, crepes and biscuits); mixtures for preparing said sweet baked goods; pie fillings and other sweet fillings (including but not limited to fruit pie fillings and nut pie fillings, such as pecan pie fillings, and fillings for biscuits, cakes, pastries, confectionery products, etc., such as cream fillings); and snack bars (including but not limited to energy bars, cereal bars, nut bars and / or fruit bars).

[0211] They can also be jelly desserts, such as cream desserts and puddings. The term "cream dessert" refers to all types of pastry cream, mousse cream, diplomat cream, sibust cream, Bavarian cream, or crème brûlée. These various creams can be used as fillings. Another type of dessert can also be frozen desserts (including, but not limited to, frozen dairy desserts, such as ice cream (including regular ice cream, soft-serve ice cream, and all other types of ice cream), as well as frozen non-dairy desserts, such as non-dairy ice cream, sorbets, etc.).

[0212] Other products conventionally prepared using animal milk may also contain the pea protein of this invention to form substitutes. These products may be acidified and / or fermented with a starter culture (such as lactic acid bacteria, vegan starter culture, or thermophilic starter culture). These may include yogurt (including, but not limited to, full-fat yogurt, low-fat yogurt, and non-fat yogurt, which may be free of milk protein and lactose). The term "yogurt" also includes fresh cheese and Petit Swiss cheese. It may also include cheese substitutes such as cheese sauce, melted cheese, pressed and cooked cheese and raw cheese, soft cheese, twisted cheese, blue cheese; it may include Emmental, string cheese, ricotta, provolone, Parmesan, Munster, mozzarella, Monterey Jack, Manchego, blue cheese, Fontina, feta, Edam, Double Gloucester, Camembert, Cheddar, Brie, Asiago, and Havati. It can also include other products, such as vegetable-based butter and even French whipped cream.

[0213] Other products that may contain the pea protein of the present invention are sauces, such as vinegar sauces, mayonnaise, tomato sauce-based sauces, or syrups.

[0214] Similarly, the pea protein of the present invention can be incorporated into confectionery products (including but not limited to jelly candies, gummies, hard candies, chocolates, caramel, and chewing gum); sweetened and unsweetened breakfast cereals (including but not limited to extruded cereals, flake cereals, and puffed cereals); and cereal coating compositions for preparing breakfast cereals. These may also include sweet sauces (including but not limited to jellies, jams, nut butters (such as peanut butter), sauces, and other spreadable products).

[0215] The pea protein of this invention can also be used as a flavoring carrier or encapsulating agent.

[0216] In the context of this invention, other types of food and beverages not mentioned herein but conventionally containing one or more proteins may also be contemplated. In particular, animal feed (such as pet food) is explicitly contemplated.

[0217] Optionally, after texturization, pea protein can also be used in meat substitutes (such as emulsified sausages or hamburgers), deli substitutes (such as ham, chicken breast, or turkey substitutes (collectively referred to as "cold cuts")), or fish or seafood substitutes. It can also be used in egg substitute formulations or for the production of protein products such as tofu or fermented black beans. Texturized protein generally refers to protein that has been texturized by extrusion (i.e., particularly dry extrusion) or by texturizing plant proteins or high-moisture extrusion. Extruders can be single-screw, twin-screw, or multi-screw. In the case of twin-screw extrusion, the extrusion can be co-rotating or counter-rotating. As examples of multi-screw extrusion, planetary extruders and annular extruders can be mentioned. Other more specific technologies can include shear unit technology, microextrusion, and 3D printing. Pea protein can also be used to manufacture binder compositions for the production of meat or fish substitutes. Binders are typically used to assemble texturized protein particles to shape these substitutes, which can be achieved through compression cooking or by using, for example, PowerHeater, sold by Source Technology. ™ Assembly is performed using molding equipment.

[0218] Foods or beverages can be used, especially for specific nutritional needs, such as for specific populations, including infants, children, adolescents, adults, the elderly, athletes, and patients. These can be meal replacement formulas or complete nutritional beverages, for example, for weight management or clinical nutrition (e.g., tube feeding or enteral nutrition).

[0219] Pea protein can be used as a single protein source, but it can also be used in combination with other plant or animal proteins. These additional proteins can be hydrolyzed or non-hydrolyzed. They are typically in the form of concentrates or isolates.

[0220] The term "plant protein" refers to all proteins derived from cereals, oil plants, legumes, and tuberous plants, as well as all proteins derived from algae and microalgae or fungi, used alone or in mixtures, and selected from the same or different families.

[0221] In this application, "leguminous plants" generally refers to the dicotyledonous family of plants belonging to the order Fabales. Several leguminous plants are important crop plants, such as soybeans, common beans (especially mung beans), chickpeas, broad beans, peanuts, cultivated lentils, cultivated alfalfa, various clover species, broad beans, locust beans, licorice, and lupins. Other leguminous proteins may be selected from these leguminous plants or may be pea protein, such as the pea protein used in this invention. In this application, the term "cereal" refers to plants cultivated from grasses that produce edible grains, such as wheat, oats, rye, barley, corn, sorghum, or rice. Tubers may be carrots, cassava, konjac, potatoes, Jerusalem artichokes, or sweet potatoes. Oil-bearing seed plants are generally plants that produce seeds from which oil can be extracted. Oil-bearing seed plants may be selected from sunflowers, rapeseed, peanuts, sesame, squash, or flax. Animal proteins may be, for example, egg or milk proteins, such as whey protein, casein, or caseinate proteins. The pea protein compositions of the present invention can therefore be used in combination with one or more of these proteins or amino acids to improve the nutritional properties of the final product, such as improving the PDCAAS of the protein or providing other functionalities.

[0222] Pea protein can also be used in the production of pharmaceuticals or fermentation, such as for the production of fungal metabolites or cell culture metabolites.

[0223] The pea protein of the present invention can be used in particular for the manufacture of proteins, meat substitutes such as emulsified sausages, deli substitutes, egg substitute compositions, or cream desserts produced by dry or wet extrusion.

[0224] The invention and its advantages will now be illustrated by embodiments described in detail in the Embodiments section below. It should be noted that these embodiments are not intended to limit the invention.

[0225] Example

[0226] method

[0227] The following examples illustrate several implementation schemes. The methods used are described in detail above.

[0228] Series 1 (Examples 1 and 2, Counterexamples 1, 2, 3 and 4): Preparation of pea protein and comparative pea protein according to the present invention Effects of extraction pH and manufacturing process

[0229] Approximately 900 kg of peas were used. First, the outer fibers of the peas were separated from the seeds by crushing (mechanical separation of the shells and seeds) and peeling (sorting the shells and peeled seeds using compressed air). The crushed seeds were then ground in a high-speed grinder to produce pea flour. The pea flour was then cooled with compressed air. The pea flour was then rehydrated with water using a high-shear rotor-stator system to ensure rapid and efficient hydration. The pea suspension was then transferred to a stirred tank. Optionally, alkalization was performed using 4% sodium hydroxide at the pH (“extraction pH”) shown in the table. The ground pea suspension was fed into a centrifugal decanter (Flottweg Z3). The protein fraction was recovered in the so-called “protein overflow.” The protein fraction was adjusted to pH 5 with hydrochloric acid in the stirred tank and then pasteurized at 70°C for less than approximately 5 seconds. The protein fraction was then rapidly cooled. Immediately afterward, the pasteurized protein fraction was aliquoted into a Flottweg Z3 decanter centrifuge. The recovered protein precipitate (“protein understream”) was diluted in warm water. The precipitate was then immediately adjusted to approximately 18% dry matter content, followed by adjustment to the pH shown in the table (“heat treatment pH”) with 4% sodium hydroxide. The table reports the percentage of denatured pea protein (“denaturation %”); the denaturation percentage of pea protein in the protein understream was between approximately 43% and 57%. The pea protein was then heat-treated at 120°C for 5 seconds, followed by rapid cooling to approximately 75°C, and finally dried in a TGE brand nozzle atomizer. The pea protein powder was recovered and its functionality (gelling ability, solubility at pH 7, viscosity) was analyzed.

[0230] Example 3: Pea protein according to the present invention

[0231] Approximately 900 kg of peas were used. First, the outer fibers of the peas were separated from the seeds by crushing (mechanical separation of the shells and seeds) and peeling (sorting the shells and peeled seeds using compressed air). The crushed seeds were then ground in a high-speed grinder to produce pea flour. The pea flour was then cooled with compressed air. The pea flour was then rehydrated with water using a high-shear rotor-stator system to ensure rapid and efficient hydration. The pea suspension was then transferred to a stirred tank. The pea suspension was then fed into a separation system that uses centrifugal force to separate the components by utilizing the centrifugal force field caused by the movement of the mixture in a stationary device. This technique allows for the separation of a heavy fraction, primarily containing starch and fiber, and a light fraction, primarily containing protein. The light fraction was then passed through a typical decanter centrifuge (Flottweg Z3). The protein fraction from the decanter centrifuge (“protein overflow”) is recovered from the overflow, along with the starch-free (based on dry weight <2%) heavy phase, which is partially composed of peptides insoluble at pH below 8.5. The protein overflow is adjusted to pH 5 with hydrochloric acid in a stirred tank and then pasteurized at 77°C for less than 5 seconds. The protein fraction is then rapidly cooled. Immediately afterward, the pasteurized protein fraction is passed through a Flottweg Z3 decanter centrifuge to recover the heavy fraction (“precipitated pea protein underflow”). The starch-free heavy phase is diluted with water at a 1:1 ratio and then alkalized to pH 9.5. This fraction is then passed through a typical decanter centrifuge system (Flottweg Z3) to recover the overflow (“peptide overflow”). The heavy fraction “precipitated pea protein underflow” (approximately 30% dry matter) is diluted with the “peptide overflow” to between 18% and 20%. The mixture was then adjusted to pH 6.3 using 4% sodium hydroxide. The pea protein in the mixture had approximately 37% denatured percentage. The purified mixture was heat-treated at 120°C for 5 seconds, followed by rapid cooling to approximately 75°C. The pea protein flocculent was then dried in a TGE nozzle atomizer. The recovered pea protein powder was then analyzed (gelling ability, solubility at pH 7, viscosity).

[0232] Example 4: Pea protein according to the present invention

[0233] Approximately 900 kg of peas were used. First, the outer fibers of the peas were separated from the seeds by crushing (mechanical separation of the shells and seeds) and peeling (sorting the shells and peeled seeds using compressed air). The crushed seeds were then ground in a high-speed grinder to produce pea flour. The pea flour was then cooled with compressed air. The pea flour was then rehydrated with water using a high-shear rotor-stator system to ensure rapid and efficient hydration. The pea suspension was then transferred to a stirred tank. The pea suspension was then fed into a separation system that uses centrifugal force to separate the components by utilizing the centrifugal force field caused by the movement of the mixture in a stationary device. This technique allows for the separation of a heavy fraction, primarily containing starch and fiber, and a light fraction, primarily containing protein. This fraction was then passed again through a typical decanter centrifuge system (Flottweg Z3). The protein fraction (“protein overflow”) is recovered from the overflow, along with a starch-free (based on dry weight <2%) heavy phase containing polypeptides insoluble at pH below 8.5. The starch-free heavy phase is then diluted with water at a 1:1 ratio to achieve a dry matter content close to 10%, and then alkalized to pH 9.2. This fraction is passed through a decanter again, its overflow (“polypeptide overflow”) mixed with the protein fraction (protein overflow). The mixture is then acidified with hydrochloric acid to pH 5, pasteurized at 70°C for less than 5 seconds, and then rapidly cooled. The pasteurized protein fraction is then passed through a decanter centrifuge (Flottweg Z3) to recover a heavy fraction referred to as the “pea isolate,” which is diluted with warm water to approximately 20%. The pea isolate is then adjusted to pH 6.3 with 4% sodium hydroxide. The pea isolate was heat-treated at 120°C for less than 5 seconds, then rapidly cooled to approximately 80°C. The isolate was then dried in a nozzle atomizer. The recovered pea protein powder was then analyzed (gelling ability, solubility at pH 7, viscosity).

[0234] Counterexample 5: Comparison of pea protein

[0235] This counterexample is almost identical to Example 4, except that:

[0236] - Do not use starch-free heavy phases and discard them (do not recover peptide overflows).

[0237] - Acidify the protein overflow to pH 5 with hydrochloric acid, then pasteurize at 77°C for less than 5 seconds, followed by rapid cooling.

[0238] - Adjust the pH of the pea isolate recovered in this way to 7.7 instead of 6.3.

[0239] Then the recovered pea protein powder was analyzed (gelling ability, solubility at pH 7, viscosity).

[0240] Example 5: Pea protein according to the present invention

[0241] Approximately 900 kg of peas were used. First, the outer fibers of the peas were separated from the seeds by crushing (mechanical separation of the outer shell and pea seeds) and dehulling (sorting the outer shell and peeled pea seeds using compressed air). The pea and water suspension was continuously milled in the wet phase to obtain a milled pea suspension with approximately 20% dry matter. The suspension was then transferred to a stirred tank and adjusted to pH 9.2 by adding 4% sodium hydroxide. This milled pea suspension was fed into a decanter (Flottweg Z3). The protein fraction was recovered from the overflow. The protein fraction was adjusted to pH 5 with hydrochloric acid in the stirred tank and then pasteurized at 70°C for less than 5 seconds. Immediately afterwards, the pasteurized protein fraction was sent to a Flottweg Z3 decanter centrifuge. The recovered protein precipitate (underflow) was diluted in warm water. The precipitate was then adjusted to a dry matter content of approximately 18%, followed by pH adjustment to 6.3 with 4% sodium hydroxide, and heat-treated at 120°C for 5 seconds, then rapidly cooled to approximately 75°C. The protein was then atomized in a TGE nozzle atomizer. The recovered pea protein powder was then analyzed (gelling ability, solubility at pH 7, viscosity).

[0242] Example 6: Pea protein according to the present invention

[0243] Example 6 was repeated in the same manner as Example 3, except that the pH during the heat treatment was set to 6.65. .

[0244] In addition to the proteins in the examples and counterexamples, the gelling ability and viscosity of commercial pea protein isolates PISANE F9, C9 and M9, marketed by COSUCRA, were also reported.

[0245] [Table 1]

[0246]

[0247] As can be seen from the various embodiments, counterexamples, and characteristics of commercial products described above, the present invention makes it possible to obtain pea proteins with a unique combination of a priori incompatible properties: in fact, by using methods controlled during different manufacturing steps, the inventors have successfully obtained pea proteins that combine temperature-gelling ability with low viscosity. However, the prior art teaches that obtaining proteins with enhanced temperature-gelling ability is accompanied by a sharp increase in viscosity at room temperature. All pea proteins according to the present invention have zero residual denaturation enthalpy.

[0248] These new proteins will now be evaluated in various applications.

[0249] Evaluation of pea protein in wet extrusion according to the present invention

[0250] Prepare powder mixtures according to the formulations described in the table below, expressed in terms of the weight of the components:

[0251] [Table 2]

[0252]

[0253] The proteins tested were pea protein from Example 1 and pea protein from Counterexample 5.

[0254] The mixture is introduced by gravity into a LEISTRITZ ZSE 27MAX extruder from LEISTRITZ.

[0255] The mixture was introduced at a regulated flow rate of approximately 13.3 kg / h. Water was also introduced at a rate of approximately 15.3 kg / h. The moisture content in the extruder was approximately 56%.

[0256] Wet extrusion tests were conducted on the extruder equipped with a temperature-controlled die, a Coperion FDK750 model, consisting of two modules, each 80 cm in length and with a flow area of ​​50 mm × 15 mm. The extrusion screw rotated at 350 rpm and fed the mixture into the die. Two temperature characteristics were used in the tests.

[0257] The temperature characteristics of the extruder equipped with 15 heated cylinders are described in detail below:

[0258] [Table 3]

[0259]

[0260] The temperature profile of the extruder is detailed below:

[0261] [Table 4]

[0262]

[0263] At the exit of the mold, the textured protein produced therefrom is cut into strips approximately 10 cm long (5 cm wide and 1.5 cm thick).

[0264] The extrusion parameters for the four tests are shown below:

[0265] [Table 5]

[0266]

[0267] Observation of fibrosis of strips

[0268] To observe the fiberization of the strip, the strip is cut longitudinally in half and the two halves are pulled apart to tear it and observe the presence or absence of fibers. The torn strip can be... Figures 2 to 5 I saw it in the middle.

[0269] The CP 1 test is not conclusive: extrusion does not occur in a stable manner: the product is torn at the extruder exit, and significant separation between the center and the shell is observed, such as... Figure 3 As shown. Conversely, the tests in Example 1 showed good fibrosis ( Figure 2 ).

[0270] At lower temperatures, CP 2 tests showed that proper fiberization of the comparable mixture could be achieved at lower temperatures, but this was still limited. Figure 5 When using a mixture of proteins utilizing the present invention, this fibrillation is excellent at the same temperature profile. Figure 4 It can also be seen that for both tests, the pressure in the extruder increases with the increase of the temperature curve, but the pressure does not become so high that the test cannot be performed.

[0271] Therefore, by using the pea protein of the present invention, and in contrast to the comparative protein, different temperature profiles can be used. In particular, higher temperature profiles (curve 1 includes the region with a maximum set temperature of 150°C) can be used to provide extruded strands in a stable manner, which is impossible with the comparative protein. Furthermore, using the pea protein of the present invention allows for the observation of better fibrillation of the extruded strands, especially when using such higher temperature profiles.

[0272] Evaluation of the use of pea protein according to the invention in the manufacture of cream desserts

[0273] The protein of the present invention evaluated is the protein of Example 4. The comparative protein used is commercial pea protein (NUTRALYS sold by the applicant). ® F85M).

[0274] The proteins of the present invention and comparative proteins were evaluated in the following cream dessert recipes. In the following text, comparative cream was obtained using comparative pea protein, and "the cream of the present invention" was obtained using pea protein according to the present invention.

[0275] [Table 6]

[0276]

[0277] This formula does not contain carrageenan.

[0278] Experimental protocol :

[0279] 1 / Hydrate the protein powder in water at 2500 rpm and 55°C for 30 minutes using a Silverson mixer.

[0280] 2 / Add the remaining powder and mix at 4000 rpm for 5 minutes.

[0281] 3 / Add coconut oil and mix at 6000 rpm for 5 minutes.

[0282] 4 / Homogenize at 57°C and 100 bar

[0283] 5 / Sterilize at 130°C for 30 seconds, then cool to 75°C.

[0284] 6 / Let it cool to room temperature (20℃)

[0285] 7. Store at 4°C.

[0286] The proteins of this invention allow for a greater development of consistency in cream desserts after sterilization and cooling. However, despite this phenomenon, no processing problems were observed during mixing and homogenization.

[0287] Sensory evaluation was conducted nine days after the cream was produced.

[0288] As a point of reference, the properties will also be compared with commercial products containing dairy products that do not contain pea protein and carrageenan (such as Danone's vanilla Danette). ® The properties of ) were compared.

[0289] The conclusion report is as follows:

[0290] [Table 7]

[0291]

[0292] Therefore, using the proteins of this invention, cream desserts can be formulated to be almost as thick and firm as commercially available cream desserts, even though the latter contain milk proteins known to generally gel stronger than pea protein, and carrageenan (a thickener known to significantly increase the texture when added to cream). This results in a cream texture in the mouth and on a spoon that is very similar to that of commercially available products.

[0293] Furthermore, both the cream dessert of the present invention and the comparative cream dessert have a satisfying taste.

Claims

1. A functionalized pea protein, according to test A, wherein the functionalized pea protein has a gelling ability of at least 400 Pa, preferably in the range of 500 Pa to 2000 Pa, and most preferably in the range of 600 Pa to 1800 Pa, and a measured viscosity of less than 0.65 Pa·s, said viscosity being measured at 15% by weight of dry matter, a shear rate of 40 s⁻¹, and a temperature of 20°C.

2. The functionalized pea protein according to claim 1, characterized in that, According to Test A, the functionalized pea protein has a gelling ability in the range of 600 Pa to 1000 Pa, or 1000 Pa to 2000 Pa, or 1100 Pa to 1800 Pa.

3. The functionalized pea protein according to any one of claims 1 to 2, characterized in that, The functionalized pea protein has a pH range of 6.0 to 6.

8.

4. The functionalized pea protein according to any one of claims 1 to 3, characterized in that, The functionalized pea protein has a viscosity of less than 0.5 Pa·s (15% dry matter, 40 s⁻¹, 20 °C), advantageously less than 0.4 Pa·s, preferably 0.05 Pa·s to 0.4 Pa·s, and preferably 0.1 Pa·s to 0.4 Pa·s.

5. The functionalized pea protein according to any one of claims 1 to 4, characterized in that, According to test C, the solubility of the functionalized pea protein at pH 7 is less than or equal to 49%, preferably less than or equal to 45%, and more preferably 20% to 40%.

6. The functionalized pea protein according to any one of claims 1 to 5, characterized in that, The functionalized pea protein has a protein content ranging from 80% to 90%.

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

8. Use of the functionalized pea protein according to any one of claims 1 to 7 in the manufacture of food or beverages, particularly proteins structured by dry or wet extrusion, meat substitutes such as emulsified sausages, deli substitutes, egg substitute compositions, or cream desserts.

9. A method for manufacturing functionalized pea protein according to any one of claims 1 to 7, characterized in that, The method includes: - Provides an aqueous pea protein composition comprising pea peptides and pea protein precipitated at isoelectric pH, wherein the peptides are insoluble at pH below 8.5, and the pea protein in the aqueous composition has a denaturation percentage of less than 65%. - Adjust the pH of the aqueous composition to a range of 6.0 to 6.

8. - The mixture is heat-treated to obtain the functionalized pea protein.

10. The manufacturing method according to claim 9, characterized in that, The aqueous pea protein composition provided comprises: - Suspend pea flour in water - Separate the suspension to provide insoluble fractions and a primary aqueous solution containing residues of suspended pea peptides. - Separate the primary aqueous solution to provide a purified primary aqueous solution and a residue containing pea polypeptides. - Suspend at least some of the residues containing pea peptides in water with a pH greater than 8.5 to form an alkaline suspension. - Separate the alkaline suspension to provide a residual solids fraction and a secondary aqueous solution containing the pea polypeptide. - Adjust the primary aqueous solution to an isoelectric pH to form a suspension of precipitated pea protein. - Separate the precipitated pea protein from the suspension of the precipitated pea protein. - The precipitated pea protein is mixed with the secondary aqueous solution to form the aqueous pea protein composition.

11. The method according to claim 10, characterized in that, The isoelectric precipitation step includes a pasteurization stage, which is carried out at a temperature below 80°C, advantageously between 45°C and 77°C, and advantageously for a duration of less than 30 seconds, preferably for 1 to 10 seconds.

12. The method according to any one of claims 10 or 11, characterized in that, The alkaline suspension is prepared at a pH greater than 8.7, or greater than 8.8, or even greater than 9.0, or at a pH greater than 9.

2.

13. The method according to any one of claims 9 to 12, characterized in that, The temperature range during the heat treatment of the mixture is 105°C to 128°C, preferably 110°C to 125°C, and most preferably 120°C, and the step advantageously lasts for 0.1 to 20 seconds, or advantageously 0.1 to 5 seconds.

14. The method according to any one of claims 9 to 13, characterized in that, The pea protein contained in the aqueous composition has the following denaturation percentage: - Less than 60%, or even less than 55%, or even less than 50% and / or - Greater than 10%, or even greater than 20%, or even greater than 30%.

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