Pea protein with milk flavor

Abstract

The present invention relates to pea proteins having milk flavor characteristics, to a method for producing said pea proteins and to the use of said proteins for the production of food or beverage products, in particular plant-based milk substitutes.

Description

Technical Field

[0001] The present invention relates to novel pea proteins with a milky taste profile. Another subject of the present invention relates to a process for producing these pea proteins. The present invention also relates to the use of said proteins in food production. Prior art

[0002] The daily protein requirement usually accounts for 12% to 20% of food intake. These proteins are provided equally by products of animal origin (meat, fish, eggs, dairy products) and plant foods (cereals, legumes, seaweed).

[0003] In developed countries, protein intake is still mainly in the form of proteins of animal origin. These proteins have good nutritional properties and interesting functional properties, which make them useful in a wide variety of food products.

[0004] However, numerous studies have shown that excessive consumption of protein from animal sources to the detriment of plant proteins is one of the reasons for the increase in cancer and cardiovascular diseases. In addition, animal proteins have many disadvantages in terms of their allergenicity (especially proteins from milk or eggs) and in terms of the environment associated with the harmful effects of intensive farming.

[0005] As a result, there is an increasing demand from manufacturers for plant-based proteins that have the beneficial nutritional and functional properties but do not have the disadvantages of animal-based proteins.

[0006] Since the 1970s, peas are the most developed legumes in Europe (mainly in France), especially as a protein source intended for animal and human food. Peas contain about 27% by weight of protein matter. The term "pea" is considered herein for its most widely accepted use, and in particular includes all wild varieties of "smooth peas" and all mutant varieties of "smooth peas" and "wrinkled peas", regardless of the purpose (human food, animal feed and / or other uses) for which the varieties are typically used. Pea protein (mainly 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 mentioned. In this method, the seeds are ground in the absence of water (a method called "dry grinding") to obtain powder. The powder is then suspended in water at room temperature and then subjected to the various steps of extracting protein.

[0007] Despite the undeniable properties of pea protein, protein extracted from pea has an undesirable flavor profile compared to other proteins, which can limit its use in certain applications. In addition, while a number of plant-based milk alternatives have been developed in recent years, including soy, rice and oat, pea plant-based milk alternatives have so far achieved little commercial success.

[0008] These flavor characteristics are unique to pea sources and are very different from other plant sources. Specifically, one of the main flavor characteristics prevalent in many proteins on the market is "pea" or "beany flavor". This flavor characteristic is undoubtedly an obstacle for many applications (especially food). A large number of studies have shown that one of the main reasons for this pea flavor characteristic is that internal lipoxygenase acts on lipids present in pea seeds (especially during protein extraction) to synthesize volatiles such as aldehydes and / or ketones (especially hexanal). Saponins and 3-alkyl-2-methoxypyrazines are also compounds that produce these unpleasant flavors ("Flavor aspects of pulse ingredients", Wibke SURoland, 2017). Gao et al.'s article "Effect of alkaline extraction pH on structure properties, solubility, and beany flavor of yellow pea protein isolate, Food Research International, May 2020, 131 (4)" also concluded the correlation between lipoxygenase inhibition and volatile reduction.

[0009] Additionally, proteins extracted from peas often have a noticeable bitter aftertaste (or "off-flavor").

[0010] For example, in order to improve the taste of vegetable proteins, a long-standing solution has been to reduce the lipid content by using organic solvents, thereby limiting the production of volatiles mentioned above. The organic solvents can be applied to the powder or directly to the obtained protein. In this regard, patent application WO2021174226 can be mentioned, which describes this technology. However, this document does not describe pea proteins with milky characteristics.

[0011] Several solutions have been developed by those skilled in the art to improve the flavor of pea protein and give it a neutral taste. The first solution is based on masking the taste by adding compounds chosen for this purpose: this solution forces users to introduce compounds into their formulations that they do not necessarily want to introduce and requires labeling in the ingredients.

[0012] Another solution is described in patent US4022919, which teaches that treating pea flour with steam makes it possible to obtain a flour with improved flavor. However, this method may be criticized because it may modify the functional qualities of the protein obtained by thermal denaturation (e.g. loss of solubility or increase in its hydration capacity) and gelatinization of the starch contained in the flour. Moreover, this document does not mention milky flavor.

[0013] Other solutions have been investigated, including but not limited to selecting pea cultivars with less lipoxygenase or pre-germinating the peas before protein extraction.

[0014] The use of long soaking of peas before grinding and protein extraction is also described. An example is patent application WO2015071499, which teaches a method comprising soaking with lactic acid bacteria for several hours at 40°C. However, these solutions are not satisfactory. This time-consuming and complex method, which consumes a lot of water due to lactic acid fermentation, does not yet allow pea proteins with a completely neutral flavor (see Table 10, where the smell and / or taste of peas can be smelled in each extract), nor can it obtain a milky taste characteristic.

[0015] It is worth mentioning patent application WO2017 / 120597, which discloses a method comprising precipitating pea protein by adding salt, washing multiple times and recovering by centrifugation. Despite the complex method using large amounts of water (up to 30 times the amount of peas), the "pea" and "bitter" flavors still exist in pea protein (see Figures 18A, B and C). Again, the document does not mention the milky characteristics of pea protein. In addition, when formulating plant-based milk substitutes, the formula of the finished product can also contain a variety of flavorings. However, in order to simplify the finished product label, people are reluctant to use these flavorings. In addition, these flavorings can be expensive. Finally, in order to achieve the desired results, the proportions of these flavorings can be difficult to control.

[0016] The applicant has also studied many other strategies for improving the taste of legume proteins (including peas). For example, the documents WO2020 / 260841 and WO2020 / 240144 can be mentioned, which describe the production of pea proteins with improved taste. In these documents, there is no mention of the milky taste characteristics of the taste of the produced pea proteins. In addition, the document WO2019 / 053387 submitted in the name of the applicant discloses a method for producing pea proteins with reduced pea flavor characteristics, the method comprising blanching the seeds at 70°C to 90°C for 2 minutes to 4 minutes, then cooling, grinding the seeds and then extracting the pea protein. The document does not describe the production of pea proteins with milky characteristics: although this flavor profile (along with other features) was presented to the panel, the panel did not use it to characterize the taste of the protein.

[0017] The use of enzymes that modify the primary structure of proteins (such as glutaminases or proteases) can also modify the organoleptic properties of proteins, including their taste; therefore, methods for producing modified pea proteins using such enzymes have been disclosed. For example, patent application US2021 / 0401022A1 can be mentioned, which discloses such a method. The milky taste characteristics are mentioned in Table 2 of the sensory evaluation. In addition to the fact that the primary structure of the obtained pea protein is modified, another disadvantage of using an enzyme such as glutaminases is that it converts glutamine and produces ammonia, which therefore reduces the amount of protein nitrogen in the pea protein.

[0018] Therefore, although it seems that research has been conducted to reduce the pea taste and bitter aftertaste of proteins, or even to try to obtain proteins with as neutral a taste as possible, it must be pointed out that none of the above documents seeks (or tries) to provide pea proteins with milky characteristics, while keeping the pea proteins unmodified and without adding any additional flavoring agents. This is also confirmed by the fact that pea proteins on the market do not have milky characteristics, as demonstrated in the Examples section.

[0019] According to this application, "flavor profile" refers to a set of flavor descriptors determined by a trained taste panel that represent the flavor characteristics of a product. A milky profile refers to a milky profile identified by a taste panel in the first three primary flavor characteristics, or even as the first or second flavor characteristic when the protein is suspended in water and tasted. A milky profile refers to a flavor characteristic associated with milk and / or yogurt.

[0020] However, there is a definite need for such proteins which can be advantageously used, for example, in plant-based milk alternatives to obtain a milkier organoleptic profile.

[0021] After extensive research, the Applicant has proposed a new production method that makes it possible to provide pea proteins with a very slight pea flavor profile and very little bitterness and milky characteristics, without the need for the addition of flavoring agents. This is clearly advantageous for the production of products such as pea plant-based milk substitutes. Moreover, since the flavor profile of the pea protein thus obtained is unique, its use, even in foods other than plant-based substitutes, makes it possible to modify the taste and flavor of the final product in whose composition pea proteins are used. According to a variant, the Applicant has also succeeded in obtaining novel pea proteins that enable an excellent texture to be achieved when used in extrusion, in particular in wet extrusion. These pea proteins are particularly advantageous for the production of meat or fish analogs. Summary of the invention

[0022] Therefore, the present invention relates to a method for producing pea protein, comprising the following steps:

[0023] a) introducing peas or ground peas into an aqueous solution at a temperature between 65° C. and 90° C. to obtain a water / pea suspension or a water / ground pea suspension;

[0024] b) heat-treating the suspension obtained in step a) at a temperature between 40° C. and 65° C. for 1 to 10 minutes;

[0025] c) in the case of a water / pea suspension, wet-milling the water / pea suspension obtained in step b) to obtain an aqueous suspension of ground peas;

[0026] d) extracting a protein fraction from the aqueous suspension of peas obtained in step b) or c) by solid-liquid separation;

[0027] e) optionally adjusting the pH of the protein fraction obtained in step d) to between 2.0 and 8.0, such as between 4.5 and 5.7;

[0028] f) heat treating the protein fraction obtained in step e) at a temperature in the range of 65° C. to 90° C. for a time in the range of 1 second to 120 seconds to form a coagulated pea protein suspension;

[0029] g) subjecting the suspension of coagulated protein obtained in step f) to solid-liquid separation to form pea protein.

[0030] Advantageously, the extraction step d) is preceded by a step d0) of cooling the suspension to a temperature below 15°C, preferably to a temperature comprised between 4°C and 14°C, for example between 10°C and 12°C.

[0031] Advantageously, the cooling step d0) is carried out by passing the aqueous suspension of ground peas through a heat exchanger.

[0032] Advantageously, the method comprises a step f1) of cooling the suspension of coagulated proteins by rapid cooling after the heat treatment step f).

[0033] Advantageously, the method comprises the step of adjusting the pH of the pea protein to a pH between 6 and 7.5, preferably between 6.5 and 7.5.

[0034] Advantageously, the method comprises the step of additionally subjecting the pea protein to a heat treatment.

[0035] Advantageously, the method comprises the step of shearing the pea protein, for example by passing it through a high pressure pump.

[0036] Advantageously, the method comprises the step of homogenizing the pea protein.

[0037] Advantageously, the method comprises the step of drying the pea protein.

[0038] Advantageously, the pH of the aqueous solution of step a) is adjusted to between 8 and 10.

[0039] Advantageously, the duration of the heat treatment of the protein fraction is between 1 second and 45 seconds, most preferably between 1 second and 10 seconds.

[0040] Advantageously, the ground peas from step a) are obtained by dry milling.

[0041] Advantageously, a fraction enriched in pea starch and / or a fraction enriched in pea fibres is recovered from the insoluble fraction resulting from the solid-liquid separation step d).

[0042] Another subject of the present invention is pea proteins obtainable by the process according to the invention.

[0043] Advantageously, the pea protein is characterized in that at least one of its first three CATA descriptors determined according to ISO 5492:2008 (en), 4.23 is a milky descriptor. Such pea protein may have a low bitterness, reduced pea flavor profile and also a milky profile. Without being bound by any theory, the applicant hypothesizes that the milky profile may be explained by the presence of volatile compounds in the pea protein of the invention, the amounts and / or proportions of these volatile compounds being different from those of known pea proteins, some of which may be produced by the steps of the method of the invention (particularly the combination of heat treatment steps) and some of which may be reduced or eliminated. Even more strikingly, the method does not require the use of organic solvents or the use of enzymes and the pea protein may also exhibit excellent functional properties, such as high solubility and / or gelling ability.

[0044] Another subject of the present invention is also the use of the pea protein for producing food or beverage products, in particular plant-based milk substitutes. DETAILED DESCRIPTION

[0045] The present invention relates to a method for producing pea protein.

[0046] Step a)

[0047] Step a) comprises introducing peas into an aqueous solution. The peas used in step a) may have previously undergone steps well known to those skilled in the art, such as in particular cleaning (removal of unwanted particles such as stones, dead insects, soil residues, etc.) or even removal of the outer fibers (outer cellulose shell) of the peas by a well-known step called "shelling". Therefore, the "peas" in step a) refer to whole peas or pea cotyledons, from which the shell has been preferentially removed. Alternatively, ground peas (i.e. pea flour) can be used, which are generally obtained by dry grinding. Beforehand, the unpeeled peas, peeled peas or pea cotyledons may be subjected to a roasting step, i.e. the legume seeds are subjected to a dry heat treatment. The dry heat treatment may be the dry heat treatment in patent application WO2020 / 260841.

[0048] The aqueous solution may be water, and may also contain additives such as antifoaming agents or bacteriostatic compounds.

[0049] In particular, the weight ratio of the amount of peas to the amount of aqueous solution in step a) may be between 0.5 and 2.

[0050] The temperature of the aqueous solution is between 65°C and 90°C. Heating can be performed using any means known to those skilled in the art, such as an immersion heat exchanger. Preferably, the temperature is between 70°C and 80°C or even about 75°C.

[0051] The pea / water suspension or ground pea / water suspension is obtained by introducing peas or ground peas into a preheated aqueous solution.

[0052] Alternatively, the pH of the aqueous solution of step a) is adjusted to between 8 and 10. This adjustment can be carried out by adding a base such as sodium hydroxide, lime or potash, preferably sodium hydroxide. According to another variant, the pH is not adjusted in this step.

[0053] Step b)

[0054] The method further comprises heat-treating the suspension obtained in step a) at a temperature between 40° C. and 65° C. for 1 to 10 minutes. The suspension may be heated or cooled to this temperature. Alternatively, the suspension is not subjected to any heating but is directly heated to the temperature at which the aqueous suspension is mixed with the peas or ground peas. Preferably, the heat-treatment temperature is between 40° C. and 60° C., or even between 45° C. and 55° C. Preferably, the heat-treatment is carried out for 2 to 4 minutes.

[0055] Step c)

[0056] If peas are used during step a), the method comprises a step c) of wet milling the pea / water suspension treated in step b) to obtain an aqueous suspension of ground peas. Preferably, the method is carried out using peas, and the wet milling step c) is carried out by continuously passing through one or more mills to obtain an aqueous suspension of ground peas. The one or more mills can be any type of mill suitable for wet milling, such as a wet ball mill, a wet cone mill, a wet spiral mill or a wet mill equipped with a rotor-stator system. According to a variant, the mill can be a mill used in the example of document WO2019 / 053387 in the name of the applicant. In a variant in which the mill is of the rotor-stator type, this type of mill can be continuously ground by passing the water / pea suspension through the mill. According to a preferred sub-variant, the method combines two cutting stages (pre-cutting and then cutting), using a different rotor-stator mill for each of these cuts. Pre-cutting and cutting can be performed sequentially, or alternatively, cutting can be performed after pre-cutting and storing the treated pea / water suspension. Such a mill is disclosed in document WO2019 / 158589. Optionally, dilution with water can be performed during or at the end of this step to form an aqueous suspension of ground peas. Alternatively, during the grinding process, water is added continuously or discontinuously to dilute the aqueous suspension. Typically, the solids content of the ground aqueous suspension ranges from 10% to 30%, for example 15% to 25%.

[0057] Step d)

[0058] Step d) of the method consists of the following process: extracting components from an aqueous suspension of ground peas, and specifically extracting a protein fraction from an aqueous suspension of peas by solid-liquid separation. Alternatively, before the solid-liquid separation stage, a stage of adjusting the pH of the aqueous suspension of ground peas can be performed. Therefore, the solid-liquid separation can be performed after the pH of the aqueous suspension of peas is adjusted to a range of 6 to 9, preferably 8 to 9, most preferably 8.5 to 9. This pH adjustment stage can be performed in a stirred tank. This stage can be shorter or longer and lasts from 1 minute to 240 minutes, for example, typically 5 minutes to 60 minutes. For pH adjustment, any type of acid and / or base, organic or inorganic, or a mixture thereof can be added. Examples of acids that can be used include hydrochloric acid, sulfuric acid, citric acid, or mixtures thereof. As examples of bases, sodium hydroxide, potassium hydroxide or lime and mixtures thereof can be mentioned. The addition of base or acid and pH measurement can be performed online. The base and / or acid can be in the form of an aqueous solution. Advantageously, before this solid-liquid separation, and preferably before the extraction step d), or even more preferably before the extraction step d) and after the heat treatment step b) or the wet milling step c), the aqueous suspension of ground peas is cooled to a temperature below 15° C. This temperature may especially be between 4° C. and 14° C., for example between 10° C. and 12° C. This cooling step d0) can be carried out using known techniques, for example by passing the aqueous suspension of ground peas through a heat exchanger.

[0059] Generally speaking, the protein fraction is the soluble part of the aqueous suspension, while the starch- and fiber-rich fraction is the insoluble part. It is also possible to separate more than two insoluble fractions, and for example to recover a first insoluble fraction rich in starch and a second insoluble fraction rich in fiber. Therefore, according to a variation of the method, a starch-rich fraction and / or a fiber-rich fraction are recovered from the insoluble fraction produced by the solid-liquid separation step d). Starch-rich fractions and fiber-rich fractions generally refer to fractions containing at least 50% starch or fiber. Methods for quantitative starch and fiber are known to those skilled in the art, and specific methods are indicated later in the specification. These fractions are usually recovered by known separation methods. Solid-liquid separation can be carried out in particular with a decanter (especially a centrifugal decanter), a centrifuge or a hydrocyclone by at least one separation step. The method can also recover one or more fiber-rich fractions / or starch-rich fractions removed from the suspension, and recover protein fractions useful for the remainder of the method of the present invention.

[0060] Step e)

[0061] The method also optionally includes step e), wherein the protein fraction is adjusted to a pH which may optionally be the isoelectric pH of the protein. The isoelectric pH refers to a pH close to the pH at which the net charge of the protein in the protein fraction is zero. The pH may be adjusted to a pH between 2.0 and 8.0, such as between 4.5 and 5.7, or even between 4.8 and 5.2. The pH may 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) may be carried out in a stirred or unstirred tank. It may be longer or shorter and last from 1 minute to 240 minutes, such as typically from 5 minutes to 60 minutes. The addition of this base or acid and the pH measurement may be carried out online, and the acid may be in the form of an aqueous solution.

[0062] Step f)

[0063] The method also includes step f), that is, heat treating the protein fraction at an optionally adjusted pH. This step includes a stage of heating a suspension of coagulated protein. This stage is carried out in a temperature range of 65°C to 90°C to form a suspension of coagulated protein. It can be carried out for 1 second to 120 seconds, preferably 1 second to 45 seconds, and most preferably 1 second to 10 seconds. A heat exchanger is usually used for this heating. It can be a type using an indirect heating principle or a direct heating principle, usually by steam injection. Preferably, heating is carried out by steam injection. Advantageously, the heat treatment step f) includes a heating stage, followed by a stage of cooling the suspension of coagulated protein. In a variation in which the heat treatment step f) includes a stage of cooling the suspension after the stage of heating the suspension of coagulated protein, the cooling stage is preferably achieved by rapid cooling referred to as "rapid cooling", thereby causing immediate cooling. At the end of this stage, the temperature range can be 60°C to 75°C, for example, between 64°C and 70°C. This rapid cooling is achieved by applying a vacuum to the suspension of coagulating protein, the vacuum applied being determined based on the cooling temperature selected.

[0064] The Applicant believes that the first steps b) and f) of the heat treatment are essential for the method of the invention. Without being bound by any theory, one hypothesis is that pea protein may contain a certain proportion of volatile compounds, which leads to giving it a milky taste characteristic. It is likely that these first heat treatment steps b) and f) carried out at a specific temperature produce this specific flavor profile, different from the flavor profile of known pea proteins. These steps may lead to a higher concentration of certain volatile compounds compared to known pea proteins, and, conversely, to a lower concentration of certain other volatile compounds. Together they can produce this milky taste profile without requiring changes in, for example, the primary structure of the protein.

[0065] Step g)

[0066] In the remainder of the process of the invention, pea protein is separated from the suspension of coagulated protein in step g). This solid-liquid separation can be carried out using the separation device shown in step e). The pea protein formed during this step mainly comprises the protein of the solid fraction separated from the liquid fraction. The liquid portion generally contains other proteins, albumin and soluble carbohydrates soluble at the isoelectric pH. The recovered solid portion contains pea protein and is generally a concentrated aqueous suspension of pea protein. The recovered solid portion has a solid content of generally 25% to 50% or even 30% to 40%. The composition by weight of pea protein can vary and will generally comprise mainly protein (particularly in the form of globulin), but also starch, lipids, fiber and / or sugar. The solid portion can be diluted with water for easier handling in subsequent optional steps.

[0067] Additional Steps

[0068] At the end of this step g), the method generally comprises a step g') of adjusting the pH of the pea protein to a pH range of 6 to 7.5, generally 6.5 to 7.5. This step can be performed by adding an inorganic or organic base, such as by adding sodium hydroxide. The pH is generally increased by adding an alkaline aqueous solution.

[0069] Preferably, the method comprises a step of additional heat treatment of the pea protein. In this step, the temperature and time conditions can vary greatly, for example 70°C to 140°C and last from 0.1 seconds to several minutes. According to a first variant of the additional heat treatment step, the temperature range is 70°C to 90°C and its duration ranges from 0.1 seconds to 30 minutes. According to a second variant of the additional heat treatment step, the temperature range is 90°C to 110°C and its duration ranges from 0.1 seconds to 5 minutes. According to another variant, the additional heat treatment step is carried out at a temperature of 110°C to 140°C for 0.1 seconds to 30 seconds, preferably 0.2 seconds to 15 seconds, for example 0.3 seconds to 10 seconds. The purpose of this step can be to functionalize and / or sterilize the pea protein. In order to carry out the additional heat treatment step, the pea protein can be in the form of an aqueous dispersion, preferably with a solid content in the range of 10% to 25%, for example 15% to 20%. Advantageously, the method of the present invention comprises a step f1) of cooling the pea protein after the additional heat treatment step. According to a preferred variant, the cooling step is achieved by rapid cooling. At the end of this step, the temperature may range from 60°C to 100°C, for example between 70°C and 90°C. Similarly, the rapid cooling step is performed by applying a vacuum to the aqueous dispersion of pea protein, the applied vacuum being determined based on the selected cooling temperature.

[0070] Regarding the functionality of pea proteins, they can be changed by heat treatment. Specifically, they may be affected by the pH selection of the composition subjected to additional heat treatment. For example, when the aqueous dispersion of pea proteins has a neutral pH during heat treatment, the solubility of the pea proteins obtained after this heat treatment is higher than the solubility of pea proteins heat-treated at a slightly lower pH. Similarly, when the aqueous dispersion of pea proteins has a neutral pH during heat treatment, the gelling ability of the pea proteins obtained after this heat treatment may be lower than the gelling ability of pea proteins heat-treated at a slightly lower pH. This is reflected in the Examples section below.

[0071] According to a variant of the method, it comprises a step of shearing the pea proteins, for example by passing an aqueous dispersion of the proteins through a high-pressure pump. As an example of a high-pressure pump, mention may be made of the high-pressure pumps sold by Silverson, also called "high shear mixers", such as those of the UHS series. Preferably, the shearing step is carried out by means of a high-pressure pump.

[0072] The shearing step may be performed before or after the heat treatment and / or pH raising step.

[0073] According to another variant, the method comprises a step of homogenizing the pea protein.

[0074] Any type of homogenizer can be used to perform the homogenization step. According to the present invention, it relates to an apparatus comprising a high-pressure pump and a homogenizing head, wherein the apparatus is designed so that the product to be homogenized passes through the homogenizing head under pressure. The homogenizing head consists of a reduced diameter opening, typically comprising a seat, a valve and an impact ring. Thus, passing an aqueous dispersion of pea protein through a homogenizer can homogenize the pea protein. Homogenization can be low-pressure homogenization, high-pressure homogenization or ultra-high-pressure homogenization. Depending on the homogenization technology used, the homogenization pressure can vary in a wide range of 1 bar to 1000 bar, for example 20 bar to 800 bar. According to one variant, the homogenization pressure ranges from 20 bar to 200 bar, for example 50 bar to 150 bar. According to another variant, the homogenization pressure ranges from 200 bar to 800 bar, for example 300 bar to 800 bar. According to one variant, homogenization is a single-action homogenization. According to another variant, the homogenization is a multi-action homogenization, for example a double-action homogenization. Homogenizers such as those sold by GEA or TetraPak may be used.

[0075] The homogenization step may be performed before or after the heat treatment and / or pH increase step.

[0076] The method according to the invention may also include a step of drying the pea protein. Typically, the drying step is performed to achieve a solid content of greater than 80%, preferably greater than 90%, and most preferably greater than 94% by solid weight relative to the weight of the pea protein. To this end, any technique familiar to those skilled in the art may be used, such as freeze drying, flash drying or drying drum drying, or atomization. The method may also include a step of grinding or micronization. Atomization is a preferred technique, in particular multi-effect atomization. The pea protein may be in powder form, the particle size d50 of which may vary greatly, for example from 10 μm to 500 μm, typically from 50 μm to 150 μm.

[0077] "Pea protein" refers to a pea extract that can be produced according to the method of the present invention, whose protein consists of pea protein. Generally speaking, the protein content is 60% or more by weight, advantageously 80% or more, for example 80% to 95%, especially 80% to 90%. The protein content is the N6.25 content calculated using the Dumas method. Obviously, it usually contains other small amounts of ingredients other than protein, such as starch, lipids, fiber and / or sugars. Generally, the total starch content in the pea protein produced according to the method of the present invention ranges from 0% to 20%, for example 0% to 10%, especially 0.5% to 5%. The total starch content can be measured using the AOAC 996.11 method. Generally speaking, the total fiber content can be 0% to 20%, for example 1% to 18%, especially 2% to 10%. This content can be determined by the AOAC 2017.16 method. Generally speaking, the total lipid content is 0% to 15%, for example 1% to 10%. The total lipid content can be determined by the AOAC 996.06 acid hydrolysis method. The sugar content may range from 0% to 10%, typically from 0.5% to 5%.The sugar content may be determined by high performance liquid chromatography (HPLC).

[0078] Pea protein may have a milky flavor profile. Without being bound by any theory, this may be explained by a higher concentration of certain volatile compounds and, conversely, a lower concentration of certain other volatile compounds. According to one embodiment, the 3-methylbutanal content is less than 3,000 ppb, such as less than 2,500 ppb. According to another embodiment that may be combined with the preceding embodiment, the benzaldehyde content is less than 60 ppb, such as less than 50 ppb.

[0079] The properties of pea proteins can vary widely, depending on the process parameters described previously and as shown in the Examples section.

[0080] According to one embodiment, the solubility of dry pea protein at pH 7 ranges from 10% to 99%. The solubility may have all intermediate amounts (i.e., 11%, 12%, 13% ... 97%, 98%, 99%), and a person skilled in the art will know, based on the method instructions given above and in the Examples section, how to modify the parameters of the method within the indicated ranges in order to achieve the desired solubility. Advantageously, the solubility of dry pea protein ranges from 5% to 100%, in particular from 40% to 95%. According to a first variant, the solubility ranges from 75% to 100%, for example from 80% to 95%. According to a second variant, the solubility ranges from 40% to 75%.

[0081] Solubility: Test A

[0082] Use the following test A method for determining solubility:

[0083] Measuring solubility in water

[0084] The measurement is based on diluting the sample in distilled water, centrifuging and analyzing the supernatant.

[0085] program :

[0086] 150 g of distilled water at 20°C ± 2°C are introduced into a 400 ml beaker, mixed with a magnetic stirring bar and precisely 5 g of the sample to be tested are added.

[0087] The pH was adjusted to the desired value with 0.1 N NaOH or HCl (pH 7), or without pH adjustment.

[0088] The total water content is 200 g.

[0089] Mix at 1000 rpm for 30 minutes and centrifuge at 3000 g for 15 minutes.

[0090] 25 g of supernatant was collected.

[0091] Introduce into a previously dried and weighed crystallizer.

[0092] Place in an oven at 103°C ± 2°C for 1 hour.

[0093] It was then cooled to ambient temperature in a desiccator (with desiccant) and weighed.

[0094] The soluble dry matter content is expressed in weight % and is given by the following formula:

[0095] [Formula 1]

[0096]

[0097] in:

[0098] P = weight of sample (in g) = 5 g

[0099] m1 = weight of the crystallizer after drying (in g)

[0100] m2 = weight of the empty crystallizer (in g)

[0101] P1 = weight of sample collected (in grams) = 25 g

[0102] According to one embodiment, the dried pea protein may have a gelling capacity. The gelling capacity may range from 1 Pa to 500 Pa, such as 100 Pa to 500 Pa.

[0103] Gelling ability: Test B

[0104] 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 state. The term "gel strength" may also be used. In order to quantify this gelling ability, it is therefore necessary to produce this network and assess its strength. To perform this quantification, Test B is used in the present invention, which is described as follows:

[0105] 1) Dissolve the protein composition to be tested in water at 15% + / - 2% solids and pH 7 at 60°C ± 2°C;

[0106] 2) Stir at 60℃±2℃ for 5 minutes;

[0107] 3) Cool to 20°C ± 2°C and stir at 350 rpm for 24 hours;

[0108] 4) Suspension is performed using a controlled stress rheometer equipped with concentric cylinders;

[0109] 5) Measure the elastic modulus G' and the viscous modulus G" by applying the following temperature profile:

[0110] a. Stage 1: stabilization at 20°C ± 2°C and measurement of parameter G'1 after heating from 20°C ± 2°C to 80°C ± 2°C within 10 minutes;

[0111] b. Stage 2: Stabilize at a temperature of 80°C ± 2°C for 110 minutes; c. Stage 3: Cool from a temperature of 80°C ± 2°C to a temperature of 20°C ± 2°C within 30 minutes, and measure G'2 after stabilization at 20°C ± 2°C;

[0112] 6) Calculate the gelling capacity which is equal to G'2 - G'1.

[0113] In a preferred manner, the controlled stress rheometers are selected from the models DHR 2 (TA, Instruments) and MCR 301 (Anton Paar), with spindles of the concentric cylinder type. They are equipped with a temperature regulation system based on the Peltier effect. In order to avoid evaporation problems at high temperatures, liquid paraffin is added on top of the sample.

[0114] For the purposes of this invention, a "rheometer" is a laboratory machine used to measure the rheological properties of a fluid or gel. It applies forces to a sample. Typically, due to the characteristic small size (the mechanical inertia of the rotor is very small), it allows fundamental studies of the mechanical properties of liquids, gels, suspensions, pastes, etc. in response to the applied forces.

[0115] The so-called "controlled stress" model makes it possible to determine, by applying a sinusoidal stress (oscillating mode), the intrinsic viscoelasticity values ​​of a substance, which depend in particular on time (or angular velocity ω) and temperature. In particular, this type of rheometer makes it possible to obtain the complex modulus G*, which in turn makes it possible to obtain the modulus G' or elastic part and G" or viscous part;

[0116] The first three steps consist of the following process: The protein is resuspended in water, using precise conditions that maximize subsequent measurements.

[0117] The water selected is preferably reverse osmosis water, but drinking water may also be used.

[0118] Its temperature is 60°C ± 2°C during the initial resuspension (steps 1 and 2), and then 20°C ± 2°C after 24 hours of solubilization and cooling before measurement (step 3). In general, and unless otherwise stated, when a temperature is given in this specification, it always includes a variation of ± 2°C, such as 20°C ± 2°C or 80°C ± 2°C.

[0119] A determined 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 is used, such as a beaker and a stirring rod. A 50 mL volume is stirred at 350 rpm for at least 10 hours at room temperature. Generally speaking, and unless otherwise stated, 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 using a pH meter and an acid-base reagent, as is well known in the prior art.

[0120] The fourth step consists of introducing the sample into the rheometer and covering it with a thin layer of oil to limit evaporation.

[0121] During the fifth step, the following temperature regime is then applied: a. Phase 1: heating from a temperature of 20°C±2°C to a temperature of 80°C±2°C within 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 within 30 minutes.

[0122] During this protocol, the parameter G' was continuously measured and recorded.

[0123] The sixth and final step of Test B consists of utilizing this record. Two values ​​are extracted: G'1 = G' value at the beginning of stage 1 after stabilization at 20°C ± 2°C, and G'2 = G' value at the end of stage 3 after stabilization at 20°C ± 2°C.

[0124] The gelling power is equal to G'2-G'1.

[0125] According to one variation, pea protein is an enzymatically modified protein. For enzymatically modified proteins, those skilled in the art refer to proteins whose protein structure is intentionally modified by adding at least one enzyme capable of modifying the protein structure to the protein. The enzyme may be selected from proteases, peptidases, deaminases, such as those of the EC3.5.1 type, such as glutaminases, or deiminases, such as those of the EC3.5.3 type, such as peptidylarginine deiminases. It is well known that these protein modification enzymes are used to change the physicochemical and / or sensory properties of proteins. For example, it is known from the document WO2019 / 233920A1 that peptidylarginine deiminases reduce astringency, especially the astringency of rapeseed protein. If the method includes protein hydrolysis of a protein-rich fraction, this can change the degree of hydrolysis (DH) of the protein. Preferably, the degree of hydrolysis is less than 15%, advantageously less than 10%, preferably less than 6%, for example between 3% and 5%. Those skilled in the art will know how to adjust the enzymatic protein hydrolysis conditions, or even will not perform such a step to obtain the desired DH. According to a preferred embodiment of the invention, the pea proteins are not enzymatically modified by deamidation. According to a preferred embodiment of the invention, the pea proteins are not enzymatically modified by deamination. Glutaminase can also perform the deamination step. According to another preferred variant of the invention, the protein is not enzymatically modified. An advantage of the invention is that the organoleptic properties of pea proteins can be modified and in particular a milky flavor profile can be imparted thereto without even requiring enzymatic modification of the protein. According to one embodiment, the invention can also provide proteins with an unmodified primary structure.

[0126] Pea protein obtainable by the method of the present invention

[0127] Another subject of the present invention relates to pea proteins obtainable by the method of the present invention. As previously mentioned, the proteins of the present invention have a unique milky taste characteristic of pea proteins. Without being bound by any theory, the applicant assumes that this milky taste characteristic can be explained by the presence of volatile compounds in the pea proteins of the present invention, the amounts and / or proportions of these volatile compounds being different from those of known pea proteins, some of which can be produced by the steps of the method of the present invention (especially the combination of heat treatment steps), and some of which can be reduced or eliminated. Even more striking is that the method does not require the use of organic solvents or enzymatic modification of proteins to obtain this milky taste characteristic. Advantageously, the pea protein obtainable by the method of the present invention is characterized in that at least one of its first three CATA descriptors determined according to ISO 5492:2008 (en), 4.23 is a milky taste descriptor.

[0128] Pea protein with milky descriptors in the first three CATA descriptors

[0129] Another subject of the present invention relates to pea proteins, characterized in that at least one of its first three CATA descriptors determined according to ISO 5492:2008 (en), 4.23 is a milky descriptor.

[0130] Uses of Pea Protein

[0131] Another subject of the present invention is the use of the pea proteins according to the invention for producing food or beverage products, in particular plant-based milk substitutes.

[0132] In general, the pea protein of the present invention can be used in food products and beverages, which can include an amount of up to 100% by weight relative to the total dry weight of the food or beverage product, such as pea protein in an amount of about 1% by weight to about 80% by weight relative to the total dry weight of the food or beverage product. All intermediate amounts (i.e., 2%, 3%, 4% by weight, ... 77%, 78%, 79% by weight relative to the total weight of the food or beverage product) can be used, as well as all intermediate ranges based on these amounts. These food and beverage products may be suitable for vegetarians or vegetarian groups.

[0133] A particularly interesting use of the protein of the invention relates to its use in beverages having a more pleasant taste than those obtained from other commercially available pea proteins. The pea protein of the invention can be advantageously used in the production of beverages, in particular milk substitutes, or in other words milk alternatives. In addition, due to the milky taste profile produced by the ingredient, these beverages can also have a richer milky taste profile than beverages that do not contain the protein, which is an undeniable advantage for the production of plant-based milk alternatives. In addition to improving the flavor, the invention also makes it possible to obtain a softer mouthfeel than when using other pea proteins, which is advantageous for beverages and in particular for plant-based milk alternatives, since animal milk generally has a soft mouthfeel.

[0134] In beverages, the protein content of these products may vary greatly, and may also be high-protein beverages. The protein content may range, for example, from 1% to 12% by dry weight relative to the total weight of the beverage, in particular from 3% to 10% by dry weight relative to the total weight of the beverage. The beverage may be of any type, and includes plant-based substitutes for milk or milk substitutes, including barista-type milk and coffee creamer. 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 juices, and sweetened or unsweetened tea or coffee drinks); 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 preparations or "powder mixes"). It should be noted that in beverages, flavorings or masking agents are often used to reduce the pea-flavor characteristics or bitter aftertaste of protein, or to flavor the beverage. One of the advantages of the pea protein of the present invention is that using it instead of conventional pea protein makes it possible to reduce the amount of flavoring or masking agents, or even completely eliminate these ingredients from the beverage, while retaining the very pleasant taste of the beverage. These beverages may also contain hydrocolloids; however, since pea protein provides a softer texture, the hydrocolloid content can be reduced or even eliminated while retaining the soft mouthfeel.

[0135] The food products that may be involved include baked products, such as bread products (including but not limited to sourdough bread 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 the baked products; sweet baked products (including but not limited to rolls, cakes, pies, pastries, waffles, pancakes, muffins, pancakes and cookies); mixtures for preparing the sweet baked products; 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 cookies, 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).

[0136] They can also be jelly desserts, such as dessert creams, custards and puddings. Another type of dessert can also be frozen desserts (including but not limited to frozen dairy desserts, such as ice cream (including regular ice cream, soft ice cream and all other types of ice cream), and frozen non-dairy desserts, such as non-dairy ice cream, sorbet, etc.).

[0137] Other products conventionally prepared using animal milk may also contain the pea protein of the present invention to form a substitute. These products may be acidified and / or fermented with a starter (such as a lactic acid starter, a vegetarian starter, or a mesophilic starter). These may include yogurt (including but not limited to full-fat yogurt, low-fat yogurt, and skim yogurt, which may be free of milk protein and lactose). The term "yogurt" also includes fresh cheese and small Swiss cheese. It may also include cheese substitutes such as cheese sauces, melted cheeses, pressed ripe cheeses and raw cheeses, soft cheeses, rotating cheeses, blue cheeses; 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 Havarti. It can also include other products such as plant-based butters and even crème fraiche.

[0138] Other products that may comprise the pea proteins of the invention are sauces, such as vinaigrettes or mayonnaise or ketchup based sauces or syrups.

[0139] Likewise, the pea protein of the present invention can be incorporated into confectionery products (including but not limited to jelly candies, soft candies, hard candies, chocolates, caramels and chewing gum); 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. These may also include sweet sauces (including but not limited to jelly, jam, nut butter (such as peanut butter), sauces and other spreadable products).

[0140] The pea protein of the present invention can also be used as a flavoring carrier or encapsulating agent.

[0141] Other types of food and beverages not mentioned herein but which typically comprise one or more proteins are also contemplated in the context of the present invention. In particular, animal feed (such as pet food) is explicitly contemplated.

[0142] Pea protein can also be used in meat substitutes such as emulsified sausages or hamburgers, or in fish or seafood substitutes, optionally after texturization. It can also be used in egg replacement formulas or in the production of protein products such as tofu or tempeh. Textured protein generally refers to protein textured by extrusion (i.e. dry extrusion) or textured vegetable protein or high moisture extrusion. The extruder 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 ring extruders can be mentioned. Other more specific technologies can include shear cell technology, micro extrusion and 3D printing.

[0143] The food or beverage may be used in particular for special nutrition, e.g. for specific populations, such as infants, children, teenagers, adults, the elderly, athletes, sick populations, etc. These may be meal replacement formulas or complete nutritional drinks, e.g. for weight management or clinical nutrition (e.g. tube feeding or enteral nutrition).

[0144] Pea protein can be used as a single protein source, but can also be used in combination with other additional plant or animal proteins. These additional proteins can be hydrolyzed or non-hydrolyzed. These additional proteins are usually in the form of concentrates or isolates. The term "plant protein" means all proteins derived from cereals, oil-containing plants, legumes and tuberous plants, as well as all proteins derived from algae and microalgae or fungi, which are used alone or as a mixture and are selected from the same family or different families. In this application, "legumes" generally refer to the dicotyledonous plant family of the order Fabales. Several legumes are important crop plants, such as soybeans, kidney beans (especially mung beans), chickpeas, broad beans, peanuts, cultivated lentils, cultivated alfalfa, various clovers, broad beans, locust beans, licorice and lupins. Additional legume proteins can be selected from these legumes or can be pea proteins, such as the pea proteins used in the present invention. In the present application, the term "cereal" refers to a plant cultivated from a grass plant that produces edible grains, such as wheat, oats, rye, barley, corn, sorghum or rice. Tubers may be carrots, cassava, konjac, potatoes, Jerusalem artichokes, sweet potatoes. Oilseed plants are generally plants that produce seeds from which oil is extracted. Oilseed plants may be selected from sunflower, rapeseed, peanut, sesame, pumpkin or flax. Animal proteins may be, for example, egg or milk proteins, such as whey protein, casein or caseinate protein. The pea protein composition of the present invention may therefore be used in combination with one or more of these proteins or amino acids in order to improve the nutritional properties of the final product, for example to improve the PDCAAS of the protein or to provide other functionality.

[0145] Pea proteins can also be used in the production of pharmaceuticals or fermentation, for example for the production of fungal metabolites or cell culture metabolites.

[0146] The invention and its advantages will now be illustrated in the embodiments described in detail in the Examples section hereinafter. It should be noted that these examples do not limit the invention.

[0147] Example

[0148] Example 1: Pea protein according to the present invention

[0149] About 900 kg of peas are used. The outer fibers of the peas are first separated from the seeds by crushing (mechanical separation of the husk and pea seeds) and peeling (using compressed air to classify the husk and peeled seeds). Water at 75°C is used in this first part of the process: the pea seeds and water are continuously fed to a Bruynooghe immersed screw blancher. The water / pea weight ratio is about 1. All the water enters at the inlet of the blancher and the peas are immediately placed in the 75°C water. As soon as the peas come into contact with the water, the temperature of the water / pea suspension reaches about 55°C. The screw speed is set so that the peas pass through the blancher in 3 minutes and the temperature remains stable during this time. Water is added at the outlet of the blancher to give a water / pea weight ratio of about 1.5. The water / pea suspension is immediately ground by continuous wet milling, water being introduced at room temperature during the grinding process to obtain a ground pea suspension with a solid content of about 20% at a temperature of about 35°C. The ground pea suspension was adjusted to pH 8.5 by continuous in-line addition of sodium hydroxide. The ground pea suspension was cooled to about 10° C. by passing it through a plate heat exchanger and then transferred to a stirred storage tank. The ground pea suspension was fed into a centrifugal decanter (Flottweg Z3). The protein fraction (about 7% solid content) was recovered from the overflow. The protein fraction was adjusted to pH 5 with hydrochloric acid in a stirred tank and then, after a first immediate preheating step by passing it through a plate heat exchanger, heat-treated in a GEA slide at 74° C. for about 3 seconds by steam injection. The protein fraction was then rapidly cooled to 67° C. Immediately thereafter, the heat-treated protein fraction was fed to a Flottweg Z3 centrifugal decanter. The recovered protein precipitate (underflow) was diluted in hot water (80° C.) so that it could be pumped. The precipitate was then immediately adjusted to about 15% solid content and adjusted to pH 7 with sodium hydroxide. The pea protein floccules were heat treated at 130°C for 5 seconds and then immediately cooled to about 75°C by rapid cooling. The pea protein floccules were passed through a 200 bar high pressure homogenizer and then atomized in a TGE nozzle atomizer. The recovered pea protein powder was then analyzed.

[0150] Example 2: Pea protein according to the present invention

[0151] About 900 kg of peas are used. The outer fibers of the peas are first separated from the seeds by crushing (mechanical separation of the husk and pea seeds) and shelling (classification of the husk and peeled pea seeds using compressed air). Water at 75°C is used in this first part of the process: the peas and water are continuously fed to a Bruynooghe immersed screw blancher. The water / pea weight ratio is about 1. All the water enters at the inlet of the blancher and the peas are immediately placed in the 75°C water. As soon as the peas come into contact with the water, the temperature of the water / pea suspension reaches about 52°C. The screw speed is set so that the peas pass through the blancher in 3 minutes and the temperature remains stable during this time. Water is added at the outlet of the blancher to give a water / pea weight ratio of about 1.5. The water / pea suspension is immediately ground by continuous wet milling, water being introduced at room temperature during the grinding process to obtain a ground pea suspension with a solid content of about 20% at a temperature of about 35°C. The ground pea suspension was adjusted to pH 8.5 by continuous in-line addition of sodium hydroxide. The ground pea suspension was cooled to about 10° C. by passing it through a plate heat exchanger and then transferred to a stirred storage tank. The ground pea suspension was fed into a centrifugal decanter (Flottweg Z3). The protein fraction (about 7% solid content) was recovered from the overflow. The protein fraction was adjusted to pH 5 with hydrochloric acid in a stirred tank and then, after a first immediate preheating step by passing it through a plate heat exchanger, heat-treated in a GEA slide at 74° C. for about 3 seconds by steam injection. The protein fraction was then immediately cooled to 67° C. by rapid cooling. The heat-treated protein fraction was then immediately fed to a Flottweg Z3 centrifugal decanter. The recovered protein precipitate (underflow) was diluted in hot water (80° C.) so that it could be pumped. The precipitate was then immediately adjusted to a solid content of about 15% and adjusted to pH 6.5 with sodium hydroxide. The pea protein flocs were heat treated at 120°C for 10 seconds and then cooled to about 75°C by rapid cooling. The protein flocs were atomized in a TGE nozzle atomizer. The recovered pea protein powder was then analyzed.

[0152] Other Implementations

[0153] Other embodiments of the invention were carried out and made it possible to obtain pea proteins having properties similar to those of Example 1. For simplicity, the following table lists the differences relative to Example 1 (same means no difference).

[0154] [Table 1]

[0155]

[0156] analyze

[0157] [Table 2]

[0158] nature Example 1 Example 2 Dry Ingredients 94.3％ 94.8％ N6.25 Protein (% dry basis) 85.6％ 87.3％ Solubility (pH 7) 88.6％ 38.3％ Gelating capacity (Pa) 5 198 Viscosity 5s-1 0.08 0.50 Viscosity 40s-1 0.06 0.10

[0159] Commercial pea protein

[0160] In addition to the pea protein of the present invention (Example 1), the following commercially available pea proteins were also subjected to sensory evaluation:

[0161] · S85F( Freres

[0162] · C9(Cosucra)

[0163] · 870(Puris)

[0164] · (ADM)

[0165] Comparison of pea protein - Background Art Protein of WO2019 / 053387A1

[0166] 0.8 kg of peas were used. The outer fibers of the peas were first separated from the seeds by crushing (mechanical separation of the shell and the pea seeds) and peeling (using compressed air to classify the shell and the pea seeds). The peas were placed in a container with 1.6 L of deionized water heated to 80 ° C. The temperature of 80 ° C was maintained for 3 minutes. The peas were separated from the aqueous solution by filtering through a 2 mm mesh sieve. The peas were then placed in a second container with 1.6 L of deionized water, the temperature of which was set to 7 ° C for 5 minutes. This cooling process continued until the temperature of the peas dropped to 10 ° C or lower. The peas were separated from the aqueous solution by filtering through a 2 mm mesh sieve. The peas, which weighed 1.3 kg due to water absorption, were introduced into the grinding chamber of the Robocoupe Blixer 4VV grinder. The peas were ground at maximum speed for 1.5 minutes. Then, while still grinding at maximum speed, 2.7 L of deionized water was added within 3 minutes. Finally, grinding was continued for another 0.5 minutes. The final product is a water / pea homogenate with a solid content of 20%. The homogenate is centrifuged at 5000 g for 5 minutes. The supernatant of the concentrated protein is adjusted to pH 5 and then heated to 60° C. for 10 minutes to flocculate the protein. The protein floccules are recovered by centrifugation at 5000 g for 5 minutes. The floccules are resuspended in a volume of water so that a fluid suspension can be obtained, so that the pH can be adjusted to 7 with hydrochloric acid. The floccules are then freeze-dried. A pea protein with a protein content of 81% protein / solids and 95% solids is obtained.

[0167] Comparison of pea protein - Background Art Protein of WO2020 / 260841A1

[0168] The following protein and starch extraction methods were applied:

[0169] - Place the whole peas in a ventilated oven and dry them under 4 conditions (80°C for 2 minutes, 80°C for 5 minutes, 120°C for 2 minutes and 120°C for 5 minutes)

[0170] -Separation of outer fibers and pea cotyledons

[0171] - Grind pea cotyledons with a stone mill

[0172] -Suspend the powder in water with a solid content of 17%, a temperature of 20°C ± 2°C, and a pH of 7 ± 1

[0173] - Oscillating for 30 minutes

[0174] - Separation of insoluble matter (starch and internal fibers) by centrifugation at 1,000G for 5 minutes,

[0175] - Adjust the supernatant to pH 5

[0176] - Heat to 55°C in a double-jacketed container for 20 minutes with stirring,

[0177] - Recover the protein composition by centrifugation at 5,000G for 5 minutes

[0178] - Adjust pH to 7 with 1N NaOH

[0179] - Heat treatment at 140°C for 10 seconds by direct spraying

[0180] -Spray drying

[0181] Sensory evaluation of pea protein and commercial pea protein

[0182] For each sensory test, in order to rinse the mouth and avoid saturation caused by the solution of pea protein isolate, in order to optimize the sensory analysis of this type of product, these panelists were given a specific protocol to follow. The method is described in the publication, which is incorporated by reference: Cosson, A., Delarue, J., Mabille, A.-C., Druon, A., Descamps, N., Roturier, J.-M., Souchon, I., & Saint-Eve, A. (2020). Block protocol for conventional profiling to sensory characterize plant protein isolates. Food Quality and Preference, 83, 103927.

[0183] Evaluation in water

[0184] [Table 3]

[0185]

[0186]

[0187] (1) “Sensory analysis – Methods – General guidance for establishing sensory profiles” (ISO 13299:2016)

[0188] (2) “Description of the sensory properties of a sample, but without intensity values” (ISO 5492:2008(en), 4.23)

[0189] Sensory analysis using the CATA method made it possible to define the flavor profiles of the proteins, each of which is listed in the table below:

[0190] [Table 4]

[0191]

[0192] Of the proteins tested, only the protein of the invention had the descriptor milky, and in the case of the protein of the examples this was even the first descriptor.

[0193] In addition, unlike the proteins of the present invention, some comparative proteins ( (ADM), 870(Puris), C9) Has objectionable primary descriptors, such as sulfur and / or chemical descriptors.

[0194] As for the comparative pea protein of patent application WO2019 / 053387A1, the panel did not identify the milky descriptor, and therefore the protein also lacked a milky flavor profile. Similarly, the panel did not determine a milky flavor profile for each of the four proteins produced according to patent application WO2020 / 260841A1.

[0195] Furthermore, the protein of the present invention has very little pea taste and bitterness, and therefore ranks first among the best proteins in terms of these two criteria.

[0196] Plant-based milk alternatives

[0197] preparation

[0198] Use the following recipe to make a plant-based milk alternative:

[0199] [Table 5]

[0200] Element weight% water 90.08 Pea protein of Example 1 6.20 Sunflower Oil 1.50 Powdered cane sugar 2.00 Sunflower Lecithin 0.10 Gellan Gum 0.12 total 100

[0201] Prepare nine liters of plant-based milk alternative using the following recipe:

[0202] -1. Disperse the powder in water heated to 70°C

[0203] -2. Hydrate the mixture under gentle stirring for 30 minutes (2500 rpm in a Silverson mixer) to form an aqueous protein solution

[0204] -3. Heat the oil to 65°C and disperse the lecithin, stirring for 3 minutes.

[0205] -4. Add the oil and lecithin mixture to the aqueous solution and stir vigorously for 5 minutes (6000 rpm in a Silverson mixer)

[0206] -5. Sterilization: 142°C, 5 seconds

[0207] -6. Homogenize the sterilized solution in a high pressure homogenizer at 75°C (170 bar - Stage 1

[0208] / 30 bar - Stage 2)

[0209] -7. Cool to 4℃ and bottle

[0210] Use commercially available pea protein S85F( Frères and Pea Protein P870 Produce the same type of recipe.

[0211] Texture and color

[0212] The plant-based milk substitute of the present invention has excellent texture and satisfactory color.

[0213] Sensory analysis

[0214] The plant-based milk alternatives of the present invention have less pea-flavor characteristics, less astringency, and less bitterness than plant-based alternatives prepared with commercially available pea protein.

[0215] Furthermore, the plant-based milk alternative of the present invention is the only milk alternative characterized by milk taste as the main flavor note in its flavor profile, which is explained by the milky taste profile of the protein of the present invention.

[0216] Adding masking agents to plant-based milk alternatives

[0217] The masking agent was added to the reference plant-based milk alternative in an amount of 0.08% to improve its flavor (i.e., about 1.3% by weight of S85F pea protein). However, even with the addition of this ingredient, the pea taste and bitterness of the plant-based milk substitute of the present invention is still much weaker than the pea taste and bitterness of the reference plant-based alternative containing this masking agent. In addition, the plant-based milk substitute of the present invention is still the only milk substitute characterized by a milky taste characteristic.

[0218] Powder mixture

[0219] preparation

[0220] The powder mix was produced using the following recipe:

[0221] [Table 6]

[0222]

[0223] The pea protein of the present invention and S85F( Frères) was also evaluated in a powder mix formulation.

[0224] 40 g of the powder mixture was placed in 325 mL Brand in water.

[0225] After stirring in a shaker, it was noted that in both cases, after 15-25 minutes:

[0226] No precipitation was observed

[0227] · Low and very acceptable foaming

[0228] No caking observed

[0229] Viscosity remains stable over time

[0230] Sensory analysis

[0231] The powder mix prepared with the protein of Example 1 had a milky note that was identified as being most important by the panel. It also had a pleasantly softer mouthfeel compared to the powder mix made with the commercial pea protein.

[0232] As was the case with the pea protein evaluated in water, the taste of the powder mix comprising the protein of the invention was also improved, with significantly less bitterness and pea taste, compared to the powder mix comprising the commercial product.

[0233] Addition of masking agent to the reference powder mix

[0234] A masking agent in an amount of 1.2% was added to the reference powder mixture to improve its flavor (i.e. about 1.3% by weight of pea protein). However, even with this addition, the pea taste and bitterness of the reference powder mixture were improved and could be very close to the pea taste and bitterness of the powder mixture of the present invention, which was still the only powder mixture with a milky taste characteristic. In addition, it is thus possible to provide a powder mixture with very little pea taste and bitterness without the need to add a masking agent, which would increase the cost of the formulation of the powder mixture and would also have to be labeled.

[0235] Plant-Based Pizza Cheese Alternatives

[0236] preparation

[0237] Use the following recipe to make a plant-based pizza cheese alternative:

[0238] [Table 7]

[0239]

[0240] The pea protein of the present invention and S85F( The protein was also evaluated together with commercially available pea protein from Frères.

[0241] Apply the following scheme:

[0242] Pour water into the Stephan type digester

[0243] Heating to 50℃

[0244] Add powder other than citric acid

[0245] Mix for 2 minutes at 750 rpm

[0246] Adjust pH to 4.5 with citric acid

[0247] Mix at the same speed for 1 minute and readjust the pH to 4.5 if necessary

[0248] Heating to 75°C

[0249] Pour the molten material into a silicone mold and place in a refrigerator (4°C)

[0250] Both cheeses had high melting power, an ideal property for this type of application. In both cases the cheeses shredded very well, the proteins of the invention enabling them to be shredded in longer forms than commercially available proteins.

[0251] Plant-Based Cheese Sauce Alternatives

[0252] Use the following recipe to make cheese sauce:

[0253] [Table 8]

[0254]

[0255] All cheeses produced contain an amount of pea protein and water selected to contain 6% protein.

[0256] In a Hotmix mixer equipped with a butterfly impeller, the following protocol was performed:

[0257] 1. Heat water to 55°C and mix at 300 rpm

[0258] 2. Add pea protein and mix at 300 rpm for 20 minutes

[0259] 3. Add other ingredients in powder form and mix at 300 rpm for 2 minutes

[0260] 4. Add oil and stir at 800 rpm for 2 minutes

[0261] 5. Heat to 95°C with stirring at 300 rpm and keep at this temperature for 5 minutes

[0262] 6. Cool to 43°C and stir at 300 rpm

[0263] 7. Add fermentation agent (Vega Harmony, add 0.1mL to 500mL)

[0264] 8. Place in an oven at 43°C until pH reaches 5.0

[0265] 9. Mix with Hotmix at 300 rpm for 30 seconds

[0266] 10. Package and store at 4°C

[0267] The fermentation time for all trials was about 3:25 h. The color of the cheese was quite satisfactory.

[0268] Compared to cheeses made using commercial proteins, the cheeses of the invention had an improved taste. In these tests, only cheese 2 had a texture close to that of commercial cheese spreads, the other two being slightly more fluid and less gelled.

[0269] Yogurt without texturizers

[0270] preparation

[0271] Use the following recipe to produce yogurt:

[0272] [Table 9]

[0273] Element ％ Deionized water 88.9 Pea protein of Example 1 4.3 Sunflower Oil 2.6 Cane Sugar 4.2 Fermentation Agents qs

[0274] The pea protein of the present invention and S85F( The protein was also evaluated together with commercially available pea protein from Frères.

[0275] Apply the following scheme:

[0276] 1. Heat water to 55°C

[0277] 2. Add pea protein under moderate stirring (480 rpm) and hydrate for 30 minutes

[0278] 3. Add cane sugar and stir at the same stirring speed for 5 minutes

[0279] 4. Add sunflower oil and mix at 1800 rpm for 5 minutes

[0280] 5. Place the mixture in a NIRO PANDA high pressure homogenizer heated to 60°C (105 bar in the first stage, 45 bar in the second stage (upstream), 150 bar)

[0281] 6. Pasteurize at 95°C for 10 minutes at 800 rpm stirring

[0282] 7. Cool to 42°C and add lactic acid starter

[0283] 8. Maintain at 42°C until pH reaches 4.6

[0284] 9. Through IKA Magic Lab Smoothing equipped with 4M and 2G modules

[0285] 10. Package and store at 4°C.

[0286] The texturizing agent-free yogurt of the present invention has a soft or even liquid texture. It appears creamier and softer than the texturizing agent-free yogurt produced using commercial proteins.

[0287] Conventional texturizers may also be added to improve the texture and make the yogurt more like jelly than cream.

[0288] Emulsified plant-based sausage

[0289] The plant-based sausage model was produced according to the following recipe:

[0290] [Table 10]

[0291] The pea proteins of the present invention were tested in this model.

[0292] Element ％ water 31.5 Crushed ice 25.2 Pea protein of Example 1 14.8 Gluten 3.0 Egg white 1.5 Methylcellulose 1 Sunflower Oil 16.3 Native potato starch 5.9 Salt 0.8

[0293] Prepare approximately 1.5 kg of emulsified sausage model using the following method:

[0294] In a Stephan blender cooled to 4°C and equipped with an emulsifying blade, crushed ice was dispersed in water, stirred at 750 rpm and placed under vacuum, and the speed was increased to 1500 rpm.

[0295] The oil and methylcellulose were gradually added and then mixed at the same speed under vacuum for 5 minutes.

[0296] Add protein and continue mixing at the same speed for 5 minutes

[0297] Add remaining ingredients and mix at same speed for 5 minutes

[0298] Place 140g of the mixture into a metal can and seal it.

[0299] Heat at 100°C (70% relative humidity) for 1 hour

[0300] Cool in cold water

[0301] freezing

[0302] For tasting, the jars were reheated in hot water (100°C, 20 minutes).

[0303] The sausages of the invention had a good texture and no noticeable taste, demonstrating that the proteins of the invention can be advantageously used in this application.

[0304] Evaluation of pea protein in wet extrusion

[0305] Prepared from 3% potato starch, 5% A powder mixture consisting of 150M pea fiber and 92% pea protein, the proportions are given by weight.

[0306] The pea proteins tested were those of Example 1, Example 2 and commercially available F85M Pea Protein.

[0307] The mixture was introduced by gravity into a LEISTRITZ ZSE 27MAXX extruder from LEISTRITZ.

[0308] The mixture was introduced at a regulated flow rate of about 13.3 kg / h. Water was also introduced in an amount of about 15.3 kg / h. The humidity in the extruder was about 56%.

[0309] The wet extrusion tests were carried out on this extruder equipped with a thermostatic die from Coperion, model FDK750, comprising two modules of 80 cm in length and a flow area of ​​50 mm×15 mm, the second of which was temperature regulated at 30° C. The extrusion screw rotated at a speed equal to 350 rpm and fed the mixture into the die.

[0310] The temperature profile of the extruder equipped with 15 heatable barrels is described in detail as follows:

[0311] [Table 11]

[0312]

[0313] The textured protein thus produced was cut into 10 cm strips at the outlet of the die.

[0314] The extrusion parameters for the three tests are as follows:

[0315] [Table 12]

[0316] Band observation

[0317] Regarding the tests carried out using the protein of Example 2, the HME strips obtained had very good fibrillation. These strips had many fine fibers. They also had good elasticity.

[0318] The band obtained from the protein of Example 1 was less fibrillated than the band of Example 2. The HME band obtained using the commercially available protein under the production conditions of this example was also less fibrillated than the band of Example 2.

Claims

1. A method for producing pea protein, comprising the following steps: a) introducing peas or ground peas into an aqueous solution at a temperature between 65° C. and 90° C. to obtain a water / pea suspension or a water / ground pea suspension; b) heat-treating the suspension obtained in step a) at a temperature between 40° C. and 65° C. for 1 to 10 minutes; c) in the case of a water / pea suspension, wet-milling said water / pea suspension obtained in step b) to obtain an aqueous suspension of ground peas; d) extracting a protein fraction from the aqueous suspension of peas obtained in step b) or c) by solid-liquid separation; e) optionally adjusting the pH of the protein fraction obtained in step d) to between 2.0 and 8.0, such as between 4.5 and 5.7; f) heat treating the protein fraction obtained in step e) at a temperature in the range of 65° C. to 90° C. for a time in the range of 1 second to 120 seconds to form a coagulated pea protein suspension; g) subjecting the suspension of coagulated protein obtained in step f) to solid-liquid separation to form the pea protein.

2. The method according to claim 1, characterized in that The extraction step d) is preceded by a step d0) of cooling the suspension to a temperature below 15°C, preferably to a temperature between 4°C and 14°C, such as between 10°C and 12°C.

3. The method according to claim 2, characterized in that The cooling step d0) is carried out by passing said aqueous suspension of ground peas through a heat exchanger.

4. The method according to one of the preceding claims, characterized in that It comprises a step f1) of cooling said suspension of coagulated proteins by rapid cooling after the heat treatment step f).

5. The method according to one of the preceding claims, characterized in that It comprises the step of adjusting the pH of the pea protein to a pH between 6 and 7.5, preferably between 6.5 and 7.

5.

6. The method according to one of the preceding claims, characterized in that It comprises the step of subjecting the pea protein to an additional heat treatment.

7. The method according to one of the preceding claims, characterized in that It comprises the step of shearing the pea protein, for example by passing the pea protein through a high pressure pump.

8. The method according to claim 1, characterized in that It comprises the step of homogenizing the pea protein.

9. The method according to one of the preceding claims, characterized in that It comprises the step of drying the pea protein.

10. The method according to one of the preceding claims, characterized in that The pH of the aqueous solution of step a) is adjusted to between 8 and 10.

11. The method according to one of the preceding claims, characterized in that The duration of the heat treatment of the protein fraction is between 1 second and 45 seconds, most preferably between 1 second and 10 seconds.

12. The method according to one of the preceding claims, characterized in that The ground peas of step a) are obtained by dry milling.

13. The method according to one of the preceding claims, characterized in that A pea starch-enriched fraction and / or a pea fiber-enriched fraction is recovered from the insoluble portion resulting from the solid-liquid separation step d).

14. Pea protein obtainable by the process according to one of the preceding claims.

15. The pea protein according to claim 14, characterized in that It has not been enzymatically modified by deamidation, preferably has not been enzymatically modified.

16. The pea protein according to claim 14 or 15, characterized in that At least one of the first three CATA descriptors of the pea protein determined according to ISO 5492:2008(en), 4.23 is a milky descriptor.

17. Use of pea protein according to one of claims 14 to 16 for producing a food or beverage product, in particular a plant-based milk substitute.

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