Protein extraction process with improved yield

Abstract

The present application relates to a novel process for simultaneously providing, from plant flours, preferably derived from legumes, more preferably from peas or faba beans, a protein isolate and a protein concentrate. It also relates to the compositions obtained by this process, such as a protein isolate, a protein concentrate and a starch preferably derived from legumes, more preferably from peas or faba beans, as well as the various industrial applications, including food applications, accessible with these products.

Description

Description PROTEIN EXTRACTION PROCESS WITH IMPROVED YIELD Technical Field

[0001] This application falls within the technical field of extraction and refining processes for plant proteins, in particular proteins from legumes. Previous Arts

[0002] Daily protein requirements generally range from 12 to 20% of the food ration. These proteins are provided by both animal products (meat, fish, eggs, dairy products) and plant-based foods (cereals, legumes, algae).

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

[0004] However, numerous studies demonstrate that excessive consumption of animal protein at the expense of plant-based protein is a contributing factor to the increased risk of cancer and cardiovascular disease. Furthermore, animal proteins have many drawbacks, both in terms of their allergenic potential (particularly proteins from milk and eggs) and their environmental impact.

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

[0006] Since the 1970s, the pea has been the most widely grown legume in Europe and mainly in France, particularly as a protein source for animal and human food.

[0007] To extract protein from legumes such as peas, we can first consider the "dry" protein extraction processes, preferably from peas or broad beans. The principle of these processes is to grind the seeds into a flour, which is then introduced into a turbo-separator, a device used to classify particles according to their size and density within an airflow. Turbo-separation yields a protein-enriched fraction and a starch-enriched fraction. As a guide, the protein-enriched fraction (or "light fraction") contains approximately 40-60% protein by weight and still contains between 2% and 15% starch by weight. The starch-enriched fraction... Starch (or what is also called the "heavy fraction") contains approximately 65%-70% starch by weight and still contains between 10% and 15% protein by weight. These percentages are expressed as a percentage of the total dry weight of the fraction in question.

[0008] While the protein-enriched fraction is well utilized, particularly in the food industry, the starch-enriched fraction is much less so. This latter fraction is more difficult to utilize because it is too rich in protein to be used as a starch source.

[0009] We can then discuss the so-called "wet" extraction processes of legume proteins, preferably from peas or broad beans.

[0010] One example is the so-called "isoelectric precipitation" process, such as that described in patent EP1400537. In this process, the seed is ground to obtain a flour suspended in an aqueous solvent. This suspension in water allows for the subsequent stages of protein extraction, particularly the separation of insoluble fractions rich in starches and internal fibers from the soluble fractions containing the proteins. These insoluble fractions can then be further separated, if necessary, into a starch-rich fraction and a fiber-rich fraction. This type of process then separates, by isoelectric precipitation, the proteins belonging to the globulin subgroup (approximately 80% by weight of pea proteins) and the proteins belonging to the albumin subgroup (approximately 20% by weight of pea proteins).These remain in the liquid fraction after recovery of the floc, which is mainly composed of globulins.

[0011] Another "wet process" alternative involves replacing the isoelectric precipitation step with a membrane filtration step. This membrane separation concentrates the protein fractions of globulins and albumins in the retentate, while removing some of the salts and sugars from the permeate. The article "Impact of processing on functional properties of protein products from wrinkled peas" by Fuhrmeister et al., *Journal of Food Engineering*, Volume 56, Issues 2-3, February 2003, Pages 119-129, describes such a process and compares it to isoelectric precipitation.

[0012] In these two so-called "wet" processes, the protein-enriched fraction obtained contains approximately 70% to 90% protein by weight. The starch-enriched fraction contains between 0% and 1% protein by weight. One of the main limitations of these processes is that they only generate a protein-enriched fraction, which is far too expensive for certain applications. Furthermore, water usage is a technical and societal issue; any way to reduce this usage is a definite advantage. It is therefore important to develop new processes for extracting plant-based proteins in an optimized manner, particularly to improve yields, reduce water consumption, and ultimately reduce the... number of co-products by eliminating the starch-enriched fraction (or otherwise called "heavy fraction") traditionally produced by so-called "dry" processes.

[0013] The Applicant has thus, after extensive research, developed a new process combining dry and wet extraction methods. This process yields a composition of plant proteins, preferably from legumes, and even more preferably from peas or broad beans, which allows for the simultaneous production of a protein isolate and a protein concentrate. This process therefore generates several fractions of interest, thereby maximizing the value of the processed flour. Water usage is reduced, and obtaining both an isolate and a concentrate allows the Applicant to target various markets according to their protein content requirements and cost. Furthermore, as will be exemplified below in the example section, the process described in this application optimizes the protein extraction yield from the flour and allows for adjusting the amount of starch produced relative to the concentrate and isolate fractions obtained.This innovative process therefore makes it possible to maximize the value of vegetable flours, processed by extracting as much protein as possible from the product of plant origin and thus optimize their value as well as the value of the resulting fractions in the industrial environment, particularly food as well as in the pharmaceutical and nutraceutical industries.

[0014] The inventive process will be better understood from the following description. General Description of the Invention

[0015] In the first part, the present invention relates to a process for producing a protein concentrate and a protein isolate by extraction from plant proteins, said process comprising the following steps: a. Using a plant flour having a protein content preferably of 20% to 39%, preferably of 20% to 35% by dry weight relative to the total dry weight of the plant flour, b. Separating the plant flour from step a) into a protein concentrate, and a starch-enriched fraction using a dry separation process, c.Mixing the starch-enriched fraction obtained in step b) with an enriching vegetable flour having a protein content preferably of 20% to 60%, preferably of 28% to 36% by weight, even more preferably of 30% to 35%, in a mass ratio of starch-enriched fraction / enriching vegetable flour ranging from 70 / 30 to 30 / 70, preferably from 55 / 45 to 35 / 65, d. Separation of the mixture obtained in step c) into a protein isolate, and at least one starch-enriched fraction using a wet separation process.

[0016] Secondly, the present invention relates to a protein isolate that can be obtained by an extraction process according to this application.

[0017] Thirdly, the present invention relates to a protein concentrate obtainable by an extraction process according to the present application.

[0018] Fourthly, the present invention relates to a powder mixture consisting of a starch-enriched fraction obtained by means of a dry separation process and an enriching vegetable flour having a protein content of 20% to 60% by weight, preferably 28% to 36% by weight, even more preferably between 30% and 35% by weight, in a mass ratio of starch-rich fraction / vegetable flour of 70 / 30 to 30 / 70, preferably 55 / 45 to 35 / 65.

[0019] Fifth, the present invention relates to a starch-enriched fraction containing at least 75% by weight of starch that can be obtained by a process of the present application.

[0020] In a sixth place, the present invention relates to an industrial use, particularly in animal and human food as well as in the pharmaceutical and nutraceutical industry, of a protein concentrate, a protein isolate, or a mixture thereof, which can be obtained by an extraction process according to the present application.

[0021] Seventh, the present invention relates to the industrial use, particularly in animal and human food as well as in the pharmaceutical and nutraceutical industries, of a starch-enriched fraction that can be obtained by the extraction process according to the present application.

[0022] This request will be explained in more detail in the detailed description that follows. Detailed Description

[0023] The present document relates firstly to a process for producing a protein concentrate and a protein isolate by extracting plant proteins, comprising the following steps: a. Processing a plant flour having a protein content preferably of 20% to 39% by weight, preferably of 20% to 35% by dry weight relative to the total dry weight of the plant flour; b. Separating the plant flour from step a) into a protein concentrate and a starch-enriched fraction using a dry separation process; c. Mixing the starch-enriched fraction obtained in step b) with an enriching plant flour having a protein content of 20% to 60%, preferably of 28% to 36% by weight of protein, even more preferably 30% to 35% by weight, in a mass ratio of starch-rich fraction / vegetable flour of 70 / 30 to 30 / 70, preferably 55 / 45 to 35 / 65, d. Separation of the mixture obtained in step c) into a protein isolate, and at least a starch-enriched fraction using a wet separation process.

[0024] The term "separation process" or "separation" is understood as any combination of unit steps allowing the extraction of one or more protein-containing compositions from a raw starting material, preferably plant seeds.

[0025] The term "proteins" in this application should be understood as macromolecules formed from one or more polypeptide chains consisting of a sequence of amino acid residues linked together by peptide bonds. In the specific context of the legume proteins mentioned in this application, preferably from peas or broad beans or a mixture thereof, and more preferably from peas, the present invention relates more particularly to globulins (approximately 50-60% by weight of pea proteins) and albumins (20-25% by weight of pea proteins).

[0026] For the purposes of this invention, "globulins" refers to all globulin-type proteins according to the Osborne classification, more specifically, all proteins soluble in neutral saline solutions. Pea or broad bean globulins are primarily subdivided into three subfamilies: legumins, vicilins, and convicilins. These protein families constitute, particularly for peas, the group of proteins known as precipitable proteins. This term characterizes proteins that precipitate after the application of thermal heating and / or isoelectric precipitation.

[0027] For the purposes of this invention, "albumins" refers to all albumin-type proteins according to the Osborne classification, more specifically, all proteins soluble in pure water. Albumins are typically present in pea or broad bean proteins at a level of approximately 20% by weight. They are mainly subdivided into two families called PA1 and PA2.

[0028] The term "plant proteins" should be understood as any extract containing proteins from plant sources. For the sake of clarity, this designation excludes proteins derived from eggs, milk, or animals, and includes proteins derived from plants or algae.

[0029] The term "legumes" is used here to refer to the family of dicotyledonous plants in the order Fabales. It is one of the largest families of flowering plants, third only to the Orchidaceae and Asteraceae in terms of the number of species. It comprises approximately 765 genera encompassing over 19,500 species. Several legumes are important plants cultivated crops including soybeans, beans, peas, broad beans, chickpeas, peanuts, cultivated lentils, cultivated alfalfa, various clovers, broad beans, carob, licorice.

[0030] Preferably, plant proteins, preferably from legumes, are pea proteins, broad bean proteins, or a mixture of these, even more preferably pea proteins.

[0031] The term "pea" is here considered in its broadest sense and includes in particular all varieties of "smooth pea" and "wrinkled pea", and all mutant varieties of "smooth pea" and "wrinkled pea", regardless of the uses to which said varieties are generally intended (human food, animal nutrition and / or other uses).

[0032] The term "pea" in this application includes varieties of pea belonging to the genus Pisum and more particularly to the species sativum and aestivum. The said mutant varieties are in particular those designated "mutants r", "mutants rb", "mutants rug 3", "mutants rug 4", "mutants rug 5" and "mutants lam" as described in the article by CL HEYDLEY et al. entitled "Developing novel pea starches" Proceedings of the Symposium of the Industrial Biochemistry and Biotechnology Group of the Biochemical Society, 1996, pp. 77-87.

[0033] The term "fava bean" refers to the group of annual plants of the species Vicia faba, belonging to the legume group of the family Fabaceae, subfamily Faboideae, tribe Fabeae. A distinction is made between the Minor and Major varieties. In the present invention, both wild varieties and those obtained through genetic engineering or varietal selection are excellent sources. While legume proteins, particularly those from peas or fava beans, are especially well-suited to the implementation of the invention, it is nevertheless possible to achieve this with other sources of plant protein such as oat, mung bean, lupin, potato, maize, or chickpea protein. Those skilled in the art will be able to make any necessary adaptations. For the purposes of this application, "lupin" refers to the varieties lupin (Lupinus albus L.), blue lupin (Lupinus angustifolius L.), and yellow lupin (Lupinus luteus L.).

[0034] Preferably, the vegetable protein, preferably from a legume, in the composition is selected from peas or broad beans. Even more preferably, the vegetable protein in the composition is derived from peas.

[0035] The term "flour" in the context of the present invention means any composition containing proteins obtained by grinding the seed without any additional separation process.

[0036] The protein content of the vegetable flour used in step a) of the process according to the invention, expressed in grams of protein per 100 grams of dry matter, is from 10% to 39% by weight of protein, preferably between 15% and 35%, even more preferably between 20% and 35%, even more preferably between 20% and 30%. The protein content, expressed in grams of protein per 100 grams of dry matter, can therefore be 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, or 39% by weight of protein, as well as all the ranges that can be obtained using these values ​​as lower and upper limits. Any flour intended for dry separation to increase its protein content, and thus generating a starch-enriched fraction for future use, is included in this definition.The protein content of an enriching vegetable flour, expressed in grams of protein per 100 grams of dry matter, is 10% to 60%, preferably 15% to 35% by weight of protein, even more preferably 20% to 60%, preferably 28% to 36% by weight of protein, even more preferably 30% to 35% by weight of protein. The protein content, expressed in grams of protein per 100 grams of dry matter, can therefore be 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60% by weight of protein as well as all the ranges that can be obtained using these values ​​as lower and upper bounds.The term "enriching vegetable flour" should be understood in this application as a flour used to increase the protein content by mixing with the starch-enriched fraction obtained in step b). Vegetable flours with more than 40% protein exist, such as lupin flours.

[0037] The term "protein concentrate" in the context of the present invention means any composition obtained by applying a separation process to a seed or vegetable flour, preferably a legume flour, preferably a pea or broad bean flour or a mixture thereof, preferably a pea flour, the protein content of which, expressed in grams of protein per 100 grams of dry matter, is 40% to 70%, preferably 45% to 65%, even more preferably 50% to 60%. The protein content expressed in grams of protein per 100 grams of dry matter can therefore be 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70% as well as all the ranges that can be obtained using these values ​​as lower and upper bounds.Preferably, the protein concentrate is obtained in step b) of the application process.

[0038] The term "protein isolate" in the context of the present invention means any composition obtained by applying a separation process to a seed or vegetable flour whose protein content, expressed in grams of protein per 100 grams of dry matter, is 71% to 99%, preferably 75% to 95%, even more preferably 80% to 90%, even more preferably 82% to 88%. The protein content, expressed in grams of protein per 100 grams of dry matter, may therefore be 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, as well as all the ranges that can be obtained using these values ​​as lower and upper bounds. Preferably, the protein isolate is obtained in step d) of the application process.

[0039] The first step a) of the extraction process according to the invention consists of using a vegetable flour, preferably a legume flour, preferably a pea or broad bean flour or a mixture thereof, preferably a pea flour, the protein content of which is preferably from 20% to 39%, preferably 20% to 35%, preferably from 21% to 30%, preferably from 22% to 28% by weight. The protein content expressed in grams of protein per 100 grams of dry matter can therefore be 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39% as well as all the ranges that can be obtained using these values ​​as lower and upper bounds.

[0040] Such flour can typically be obtained commercially or produced from a raw starting material, preferably vegetable seeds, even more preferably pea and / or broad bean seeds.

[0041] For the purposes of this application, "vegetable flour" means a powder obtained by reducing the particle size of a vegetable seed, typically by grinding.

[0042] The plant seed can be of any botanical origin. Preferably, the plant seed will be a pea seed or a broad bean seed, or a mixture of pea and broad bean seeds. Other plant seeds such as soybeans, lupins, and rapeseed are also suitable for the extraction process.

[0043] The seed can then be sorted to remove foreign matter such as pebbles or stem fragments, and cleaned. The seed will preferably be separated from its outer covering (typically called the "hull" in the field). The seed may then undergo one or more pretreatments such as heating or treatment with ultraviolet, aqueous soaking followed by drying, sterilization, pre-grinding, listed here in a non-exhaustive manner.

[0044] A particle size reduction process is then carried out, breaking the seed into multiple finer particles. This particle size reduction aims to obtain a defined Particle Size Distribution (PSD). This PSD is chosen to allow step b) the separation of the plant flour from step a) into a protein concentrate and a starch-enriched fraction using an optimal dry separation process. For this purpose, mills from the POITEMILL company can be used, for example, but not limited to, mills from the company POITEMILL.

[0045] The second step (b) aims to separate the vegetable flour from step (a) into a protein concentrate and a starch-enriched fraction using a dry separation process. Dry separation can be carried out, for example, by a turbo-separation process or an electrostatic separation process.

[0046] A preferred method is air classification, also known as turbo-separation. Turbo-separation uses an air classifier that operates using an upward airflow to classify the different components of flour. The starch-rich and protein-rich particles obtained after milling the grain into flour have different weights and sizes. These differences are used to separate the protein from the other parts of the flour. Air classifiers are well-known and available, for example, from equipment suppliers like Hosokawa-Alpine. An air classification process yields a protein-enriched fraction, also called protein concentrate, and a starch-enriched fraction.

[0047] It will be easy for a person skilled in the art to acquire and configure a turbo-separator to generate starch and protein enriched fractions with the desired yields and concentrations. They can, in particular, refer to the reference article J3160 v1 from the "Techniques de l'ingénieur" collection, entitled "Pneumatic Classification: Parameters of Pneumatic Classification" (Author(s): Pierre BLAZY, El-Aïd J DI D, Publication Date: June 10, 2010).

[0048] Preferably, the dry separation process of step b) will be parameterized to obtain a protein concentrate containing 40% to 70% protein, preferably 45% to 60%, and preferably 50% to 55% protein by weight. The protein content, expressed in grams of protein per 100 grams of dry matter, may therefore be 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70% by weight of protein as well as all the ranges that can be obtained using these values ​​as lower and upper bounds.

[0049] Preferably, the dry separation process of step b) will be parameterized to obtain a starch-enriched fraction containing 50% to 90% starch by weight, preferably 60% to 88% starch by weight, preferably 80% to 88% starch by weight, and even more preferably 86% starch by weight. The starch content, expressed in grams of starch per 100 grams of dry matter, could therefore be 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%. 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, as well as all the ranges that can be obtained using these values ​​as lower and upper bounds.

[0050] Even more preferably, the protein extraction yield of the dry separation process of step b), expressed as the total amount of protein in the protein concentrate relative to the total amount of protein used in a vegetable flour, preferably a legume flour, preferably a pea or broad bean flour or a mixture thereof, preferably a pea flour, is 10% to 30%, preferably 15% to 25%. The protein extraction yield of the dry separation process in step b) can therefore be 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30% and all the ranges that can be obtained using two of these values ​​as upper and lower bounds.

[0051] Step c) aims at mixing the starch-enriched fraction obtained in step b) with an enriching vegetable flour having a protein content preferably of 20% to 60%, preferably of 28% to 36%, even more preferably of 30% to 35% by weight in a mass ratio of starch-rich fraction / vegetable flour ranging from 70 / 30 to 30 / 70, preferably ranging from 55 / 45 to 35 / 65.

[0052] The mixing is carried out using equipment familiar to those in the trade, including tanks, pumps, agitators, and scales. The equipment is chosen depending on whether the compounds are to be mixed in solid or liquid form.

[0053] For solid-form mixing, equipment such as drum mixers, convective mixers, fluidized bed mixers, or static mixers is used to homogenize the powders. The resulting solid mixture must then be suspended in aqueous solution in step d1) described below.

[0054] For liquid mixing, a tank equipped with an agitator and piping for supplying an aqueous solution is sufficient. The use of a A homogenizer and a heating system are, however, possible. In these cases, the weight ratio of dry mix (enriching flour and starch-rich fraction) to aqueous solution can range from 0.1 to 2. The target dry matter content is 10% to 30% by weight of wet mix, preferably 15% to 25% by weight of wet mix, and even more preferably 17% to 22%. Step c) yields an aqueous suspension that can directly supply the following step d).

[0055] In one particular embodiment, the mixture obtained in liquid or solid form includes the addition of compounds other than the starch-enriched fraction obtained in step b) and the enriching flour. These compounds may include, for example and without limitation, flavorings or flavor modifiers, pH-modifying agents, including buffers or reagents, or salts.

[0056] The enriching vegetable flour used for this purpose has a protein content of 20% to 60%, preferably 20% to 35%, preferably 28% to 35%, even more preferably 30% to 35% by weight. The protein content of the vegetable flour used in step c) can therefore be 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60% as well as all the ranges that can be made with two of these values ​​as lower and upper bounds.

[0057] In a particular embodiment, the enriching flour has a protein content of 20% to 27% by weight. The use of enriching flour with a protein content of 20% to 27% by weight makes it possible to carry out the invention by obtaining a process which, as will be explained and exemplified in this application, generates a protein concentrate fraction, a protein isolate fraction, a starch-rich fraction, a fiber-rich fraction, and a soluble fraction containing albumin and fiber. In this inventive process, there is no longer a generation of a so-called "heavy" fraction containing starch but also proteins.

[0058] In a specific method, the enriching flour has a protein content of 28% to 35% by weight. The use of an enriching vegetable flour with a protein content of 28% to 35% by weight is even more advantageous because, as will be explained and illustrated in this application, less flour is used, the quantity of starch produced is similar to a conventional wet process, and the protein extraction yield is improved.

[0059] In a particular form, fortified flour has a protein content of 36% to 60% by weight. The use of fortified flour with a protein content of 36% to A 60% protein content by weight is similar to using a fortified flour with a protein content of 28% to 35% by weight. It also allows for a significant reduction in starch production.

[0060] The mass ratio of starch-rich fraction to enriching vegetable flour is 70 / 30 to 30 / 70, preferably 55 / 45 to 35 / 65. This ratio should be understood as the relative mass ratio of the two fractions mixed. For example, a mixture of 500 kg of a starch-rich fraction and 500 kg of vegetable flour is characterized by a starch-rich fraction / vegetable flour mass ratio of 50 / 50. This ratio could therefore be 70 / 30, 69 / 31, 68 / 32, 67 / 33, 66 / 34, 65 / 35, 64 / 36, 63 / 37, 62 / 38, 61 / 39, 60 / 40, 59 / 41, 58 / 42, 57 / 43, 56 / 44, 55 / 45, 54 / 46, 53 / 4, 52 / 48, 51 / 49, 50 / 50, 49 / 51, 48 / 52, 47 / 53, 46 / 54, 45 / 55, 44 / 56, 43 / 57, 42 / 58, 41 / 59, 40 / 60, 39 / 61, 38 / 62, 37 / 63, 36 / 64, 35 / 65, 34 / 66, 33 / 67, 32 / 68, 31 / 69, 30 / 70, as well as all the ranges accessible using two of these ratios as upper and lower bounds. Preferably, the ratio can be from 66 / 34 to 34 / 66, and even more preferably from 60 / 40 to 40 / 60.

[0061] The mixture obtained in step c) is preferably characterized by a protein content of 20% to 35%, preferably 20% to 30% by weight.

[0062] Step d) aims at separating the mixture obtained in step c) into a protein isolate and a starch-enriched fraction using a wet separation process.

[0063] The wet separation process of step d) is preferably selected from a protein precipitation process at isoelectric pH or an ultrafiltration process.

[0064] If the mixture prepared in step c) is in solid form, a first substep d1) involves suspending the mixture obtained in step c) in an aqueous solvent. The weight ratio of the quantity of mixture prepared in step c) to the quantity of aqueous solvent in substep d1) can range from 0.1 to 2. The aqueous solvent can be water and may optionally include additives such as antifoaming agents, salts, or bacteriostatic agents. The resulting aqueous suspension can then be used in the subsequent stages of the process.

[0065] In one variation, the pH of the aqueous suspension obtained in liquid form during step c) or substep d1) is then adjusted to between 8.0 and 10.0; preferably between 8.5 and 9.5. This adjustment can be made by adding a base such as sodium hydroxide, lime, or potassium hydroxide, preferably sodium hydroxide. In another variation, the pH is not adjusted at this step. Adjusting the pH optimizes protein extraction in the subsequent stages of the process. The recovery yield is thus optimized.

[0066] Substep d2) consists of extracting at least one so-called insoluble fraction from the aqueous suspension obtained in liquid form during step c) or substep d1) by solid-liquid separation, preferably by decantation or filtration. According to one variant, before carrying out the solid-liquid separation stage, it is possible to perform a pH adjustment stage for the aqueous suspension. Thus, the solid-liquid separation can take place after adjusting the aqueous suspension to a pH ranging from 6 to 9, preferably from 8 to 9, and most preferably from 8.5 to 9. This pH adjustment stage can be carried out in a stirred tank. This stage can vary in length, lasting, for example, from 1 to 240 minutes, generally from 5 to 60 minutes. To perform the pH adjustment, it is possible to add any type of acid and / or base, organic or inorganic, or mixtures thereof.Examples of acids include hydrochloric acid, sulfuric acid, citric acid, or mixtures thereof. Examples of bases include sodium hydroxide, potassium hydroxide, or lime, and mixtures thereof. The addition of base or acid, as well as pH measurement, can be performed online. The base and / or acid can be in aqueous solution. Advantageously, before this solid-liquid separation, the aqueous suspension is cooled to a temperature below 15°C. This temperature can range from 4 to 14°C, for example, from 10 to 12°C. This cooling step can be carried out using known techniques, such as passing the aqueous suspension of ground pea or broad bean seeds through a heat exchanger.

[0067] Generally, the majority of the protein fraction is present in the soluble part of the aqueous suspension of crushed pea or broad bean seeds, and the fraction rich in starch and fiber is the insoluble part.

[0068] A typical process involves the following steps in sequence: - Passing the aqueous suspension from step c) or substep d1) through one or more centrifugal separators, preferably horizontal decanters, in order to generate a protein-enriched supernatant and a decanter enriched in insoluble particles including starch and insoluble fibers. - Rectification to pH ranging from 8 to 10, preferably 9, of the decanter enriched with insoluble particles including starch and insoluble fibers. - Passing the rectified, insoluble-particle-enriched decantation to pH 8 to 10, preferably 9, through one or more centrifugal separators, preferably horizontal decanters, to generate a second protein-enriched supernatant and a second insoluble-particle-enriched decantation including starch and insoluble fibers. - Mixture of the two protein-enriched supernatants

[0069] It is also possible to separate the constituents of the insoluble fraction, and for example recover a first insoluble fraction richer in starch and a second insoluble fraction richer in fiber.

[0070] Thus, according to one variant of the process, a typical process includes the following sequence of steps: - Passing the aqueous suspension through one or more hydrocyclones allows the production of a decanter rich in starch and a supernatant rich in insoluble fibers and proteins. - Passing the supernatant rich in insoluble fibers and proteins through one or more centrifugal separators, preferably horizontal decanters, in order to generate a protein-enriched supernatant and a decanter enriched in insoluble particles including insoluble fibers. - Rectification to pH ranging from 8 to 10, preferably 9, of the decanter enriched with insoluble particles including starch and insoluble fibers. - Passing the decanter enriched with insoluble particles, rectified to pH ranging from 8 to 10, preferably 9, through one or more centrifugal separators, preferably horizontal decanters, in order to generate a second supernatant enriched with proteins and a second decanter enriched with insoluble particles including insoluble fibers - Mixture of the two protein-enriched supernatants

[0071] The term "starch-rich fraction and fiber-rich fraction," or "insoluble fraction," generally refers to a fraction comprising at least 50% by weight of starch and / or fiber. Methods for quantifying starch and fiber are known to those skilled in the art, and specific methods are described later. These fractions are recovered conventionally using known separation methods. Solid-liquid separation can be achieved, in particular, by means of at least one separation step with a decanter, such as a decanter centrifuge, a centrifuge, or hydrocyclones. The process can also recover one or more fiber- and / or starch-enriched fractions, which are removed from the suspension, and recover the protein fraction required for subsequent processing of the invention.

[0072] This step is key in differentiating it from concentrates obtained by dry separation. The protein isolate obtained at the end of this step contains between 0% and 0.5% starch by weight, unlike concentrates obtained by turbo-separation, which contain between 2% and 15% starch by weight. This step will have a direct impact on the final composition, as will be discussed later in this application.

[0073] Following substep d2), the protein isolate will enter substep d3), which aims to extract the proteins, typically globulins and / or albumins, and purify them by eliminating other soluble compounds such as sugars, soluble fibers or salts. Two pathways are particularly well known.

[0074] In a first preferred mode typically called "isoelectric pH precipitation," substep d3) consists of precipitating the proteins at their isoelectric pH followed by recovery of the precipitated proteins by decantation, centrifugation, or filtration, preferably by centrifugation. In this mode, the protein isolate undergoes an adjustment step to the protein's isoelectric pH, preferably from 4.0 to 6.0, generally from 4.5 to 5.5, preferably 5. This step can be carried out by adding inorganic or organic acid and / or base, for example, by adding hydrochloric acid and sodium hydroxide. The pH is then raised by adding a basic aqueous solution.

[0075] Even more preferably, the step of adjusting the protein to the isoelectric pH is associated with a heating step at a temperature of 40°C to 100°C, preferably from 50°C to 90°C, even more preferably from 60°C to 80°C.

[0076] It is advantageous to perform thermal flocculation of the proteins by adjusting the protein-rich fraction to a pH value corresponding to the isoelectric point (pI) of said proteins, for example, a pH value of approximately 4.5 for pea globulins. The proteins can then be flocculated at a temperature preferably between 40 and 70 °C for a duration preferably of 5 seconds to 30 minutes, preferably 10 to 30 minutes.

[0077] This flocculation time / temperature diagram allows for a protein recovery yield of 65 to 85% by weight of extracted protein / total protein. A professional will know how to adjust pH, time, and temperature to optimize this separation according to the botanical origin of the proteins.

[0078] Preferably, the isoelectric pH precipitation step of the proteins is carried out by adjusting the pH between 4 and 6, preferably 5, followed by heating to a temperature of 50°C 7.2°C to 60°C 7.2°C, preferably from 52°C 7.2°C to 58°C 7.2°C, even more preferably from 54°C 7.2°C to 56°C 7.2°C.

[0079] The contact time ranges from 10 to 30 minutes, preferably from 15 to 25 minutes, and even more preferably 20 minutes. The aim here is to separate the initial plant proteins, preferably legume, preferably pea, broad bean, or a mixture thereof, preferably pea, from the other constituents of the supernatant from step d3). It is crucial to carefully control the time / temperature parameters.

[0080] Preferably, heating is achieved by indirect steam injection, for example into a double jacket fitted to a stirred tank.

[0081] The step following protein precipitation at their isoelectric point consists of recovering the precipitated proteins by decantation, centrifugation, or filtration, preferably by centrifugation. This separates the solid fractions, which have concentrated the proteins, from the liquid fraction, called the soluble fraction, which has concentrated the sugars, particularly galacto-oligosaccharides, amino acids, peptides, particularly the Pa1b peptide, and salts.

[0082] In a preferred mode, the step of recovering precipitated proteins by decantation, centrifugation or filtration, preferably by centrifugation, also aims at recovering the liquid fraction called the soluble fraction.

[0083] This soluble fraction is typically used in animal feed or industrial fermentation. It can also serve as a basis for extracting the albumins, galacto-oligosaccharides, and / or peptides of interest it contains.

[0084] The extraction process according to the invention reduces the amount of albumins and salts contained in the soluble fraction, thus simplifying the valorization of this co-product. The reduced salt content is particularly beneficial from an ecological perspective, as it decreases salt discharge into the environment.

[0085] The soluble fractions can be purified by decantation, centrifugation, and / or filtration to extract an albumin-enriched fraction and a galacto-oligosaccharide-enriched fraction. The galacto-oligosaccharide-enriched fraction can also undergo fermentation and / or enzymatic treatment to defructosylate the galacto-oligosaccharides.

[0086] In a second preferred mode typically called "filtration", substep d3) consists of a filtration step of the protein-enriched fraction allowing the separation of the different compounds according to their size, molecular weights.

[0087] The filtration process is preferably an ultrafiltration step of the soluble fraction allowing enrichment in proteins and the removal of a permeate in order to generate a protein-enriched retentate.

[0088] Ultrafiltration refers to the membrane separation method, distinguished from microfiltration or nanofiltration by the size of the particles in suspension or solution that can pass through it. For ultrafiltration, this size ranges from 1 to 100 nanometers (nm).

[0089] Preferably, ultrafiltration is performed with a selected cutoff threshold of 5 kDa to 10 kDa (kDa meaning kilodaltons). The cutoff thresholds can therefore be 5 kDa, 6 kDa, 7 kDa, 8 kDa, 9 kDa and 10 kDa, as well as all ranges formed by these values.

[0090] Preferably, the filtration temperature is from 45°C to 60°C. The filtration temperature can therefore be 45°C, 46°C, 47°C, 48°C, 49°C, 50°C, 51°C, 52°C, 53°C, 54°C, 55°C, 56°C, 57°C, 58°C, 59°C or 60°C, as well as all the ranges formed by these values.

[0091] Preferably, the transmembrane pressure will be between 1 and 5 bar, preferably between 2 and 4 bar. Transmembrane pressure values ​​may be 1 bar, 2 bar, 3 bar, or 4 bar. Transmembrane pressure is a parameter well known to those skilled in the art; it represents the pressure difference across the ultrafiltration membrane.

[0092] Preferably, ultrafiltration is carried out in a tangential flow filtration module.

[0093] Preferably, ultrafiltration is followed by a diafiltration step. Diafiltration consists of a series of successive steps involving the concentration of the ultrafiltration retentate and the addition of water to increase its purity. Preferably, diafiltration is carried out to achieve a minimum protein content of 70% by mass relative to the mass of the retentate.

[0094] Optionally, a step d4) to rectify the pH of the proteins obtained in step d3) by isoelectric precipitation or filtration is carried out between 5.0 and 9.0, preferably between 6.0 and 8.0. This step can be performed by adding inorganic or organic acid and / or base, for example, hydrochloric acid and sodium hydroxide. The pH is generally raised by adding a basic aqueous solution.

[0095] Preferably, an optional substep d5) consists of heat treatment of the proteins obtained in steps d3) or d4). The temperature and time conditions in this step can vary widely, for example, from 70 to 140°C and from 0.1 seconds to several minutes. According to a first variant of this heat treatment step, the temperature ranges from 70 to 90°C and the duration from 0.1 seconds to 30 minutes. According to a second variant of this heat treatment step, the temperature ranges from 90 to 110°C and the duration from 0.1 seconds to 5 minutes. According to yet another variant, this heat treatment step is carried out at a temperature of 110 to 140°C for a time of 0.1 to 30 seconds, preferably from 0.2 to 15 seconds, for example, from 0.3 to 10 seconds. This step may aim to functionalize and / or sanitize the vegetable protein, preferably from legumes, and even more preferably from peas or broad beans.To carry out this heat treatment step, the vegetable protein, preferably from a legume, and even more preferably from peas or broad beans, can be in the form of an aqueous dispersion, preferably having a dry matter content of 10 to 25% by mass relative to the mass of the aqueous dispersion, for example 15 to 20%. Advantageously, The process of the invention comprises, following the heat treatment step, a cooling step of the vegetable protein, preferably a legume protein, and even more preferably a pea or broad bean protein. According to a preferred embodiment, this cooling step is achieved by flash cooling. At the end of this step, the temperature can range from 60 to 100°C, for example from 70 to 90°C. Similarly, this flash cooling step is carried out by applying a vacuum to the aqueous dispersion of vegetable protein, preferably a legume protein, and even more preferably a pea or broad bean protein, the applied vacuum being determined according to the chosen cooling temperature.

[0096] According to one variant of the process, an optional substep d6) consists of shearing the proteins obtained in steps d3), d4), or d5), for example, by passing the aqueous protein dispersion through a high-pressure pump. High-pressure pumps marketed by SPX are examples of such pumps. Preferably, the shearing step is carried out using a high-pressure pump. The shearing step can take place before or after the heat treatment and / or pH raising steps.

[0097] According to another variant, an optional substep d7) consists of the homogenization of the proteins obtained in steps d3), d4), d5) or d6).

[0098] To perform this homogenization step, any type of homogenizer can be used. According to the invention, this refers to equipment comprising a high-pressure pump and a homogenization head, in which the equipment is designed so that the product to be homogenized passes under pressure through this homogenization head. A homogenization head consists of a reduced orifice generally comprising a seat, a valve, and a shock ring. Passing the aqueous dispersion of vegetable protein, preferably from a legume, and even more preferably from peas or broad beans, through the homogenizer thus enables the homogenization of the vegetable protein, preferably from a legume, and even more preferably from peas or broad beans. The homogenization can be low-pressure, high-pressure, or ultra-high-pressure.The homogenization pressure can vary widely, ranging from 1 to 1000 bar, depending on the homogenization technique used, for example, from 20 to 800 bar. In one variant, the homogenization pressure ranges from 20 to 200 bar, for example, from 50 to 150 bar. In another variant, the homogenization pressure ranges from 200 to 800 bar, for example, from 300 to 800 bar. In one variant, the homogenization is single-effect homogenization. In another variant, the homogenization is multi-effect homogenization, for example, double-effect homogenization. Homogenizers that can be used are marketed, for example, by the company GEA or Tetra Pak.

[0099] The homogenization step can take place before or after the heat treatment and / or pH raising steps.

[0100] The process according to the invention may also optionally include an optional substep d8) of drying the proteins obtained in steps d3), d4), d5), d6) or d7).

[0101] Generally, this drying stage is carried out to achieve a dry matter content greater than 80% by weight, preferably greater than 90% by weight, and most preferably greater than 94% by weight of dry matter relative to the weight of the vegetable protein, preferably legume, even more preferably pea or broad bean or a mixture thereof, and even more preferably pea. Any technique well known to those skilled in the art is used for this purpose, such as freeze-drying, flash drying, drum drying, or spray drying. The process may also include a grinding or micronization stage. Spray drying is the preferred technology, particularly multi-effect spray drying.Vegetable protein, preferably from legume, even more preferably from peas or broad beans or a mixture of these, even more preferably from peas, may be in powder form with a particle size d50, which may vary widely, for example from 10 to 500 pm, generally from 50 to 150 pm.

[0102] In the present invention, "D50" refers to the particle size measured in micrometers separating into two number populations containing respectively 50% and 50% by number of the total number of particles in the protein composition.

[0103] To perform this D50 measurement, a laser particle size analyzer is preferably used, and even more preferably the Mastersizer 2000 from Malvern. The parameters used are as follows: Liquid application, dispersion in ethyl alcohol; Refractive index: 1.52; Absorption index: 0.1; no use of ultrasound.

[0104] The steps of the process can preferably be carried out in this precise order but, according to the needs of the person in the trade, they can be permuted in particular steps d4) to d7). Other optional steps can also be implemented, such as a pH adjustment.

[0105] In a preferred embodiment, the process for producing a protein concentrate and a protein isolate by extraction from plant proteins according to the invention consists of the following successive steps: a. Using a vegetable flour having a protein content preferably of 20% to 39%, preferably of 20% to 35% by dry weight relative to the total dry weight of vegetable flour, b. Separation of the vegetable flour from step a) into a protein concentrate and a starch-enriched fraction using a dry separation process, c. Mixing the starch-enriched fraction obtained in step b) with an enriching vegetable flour having a protein content preferably of 20% to 60%, preferably of 28% to 36% by weight, even more preferably of 30% to 35%, in a mass ratio of starch-enriched fraction / enriching vegetable flour ranging from 70 / 30 to 30 / 70, preferably from 55 / 45 to 35 / 65, d. Separation of the mixture obtained in step c) into a protein isolate, and at least a starch-enriched fraction using a wet separation process consisting of the following successive steps: d1) suspending the mixture obtained in step c) in an aqueous solvent.d2) extraction of at least one so-called insoluble fraction of the aqueous suspension obtained in liquid form during step d1) by solid-liquid separation d3) precipitation of the proteins at their isoelectric pH followed by their recovery of the precipitated proteins by decantation, centrifugation or filtration, d4) rectification of the pH of the proteins obtained during step d3) by isoelectric precipitation or filtration is carried out between 5.0 and 9.0, preferably between 6.0 and 8.0 d5) heat treatment of the proteins obtained in step d4), the temperature ranging from 70 to 140°C and the time ranging from 0.1 seconds to 30 minutes d8) drying of the proteins obtained in step d5).

[0106] A second object of the invention is a protein isolate that can be obtained by an extraction process according to the present application.

[0107] A third object of the invention is a protein concentrate that can be obtained by an extraction process according to the present application.

[0108] A fourth object of the invention is a mixture in powder form consisting of a starch-enriched fraction obtained by means of a dry separation process and a so-called enriching vegetable flour having a protein content of 20% to 60% by weight, preferably 28% to 36% by weight, even more preferably 30% to 35% by weight, in a mass ratio of starch-rich fraction / vegetable flour of 70 / 30 to 30 / 70, preferably 55 / 45 to 35 / 65.

[0109] Preferably, the starch-enriched fraction obtained by a dry separation process contains from 1% to 70% protein, preferably from 10% to 65% protein, and even more preferably from 20% to 60% protein, the protein content being expressed in grams of protein per 100 grams of dry matter. The protein content in the starch-enriched fraction may therefore be 1%, 2%, 3%, 4%, 5%, or 6%. 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36% 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51% 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66% 67%, 68%, 69%, 70% as well as all the ranges that can be obtained with two of these values ​​used as high and low limits.

[0110] A fifth object is a fraction containing at least 75% by weight of starch that can be obtained by the process according to this application.

[0111] In the so-called "dry process" of the prior art, the starch-enriched fractions contain only 70% by weight at most of starch.

[0112] The so-called "wet process" of the prior art allows the production of enriched fractions containing at least 70% by weight, but the starch obtained differs from that obtained in the enriched fractions containing at least 70% by weight that can be obtained by the process according to the present application because the latter contain a part of so-called "damaged" starch from the starch-rich fraction obtained by the dry separation process.

[0113] In the present invention, "starch" will be understood to mean a mixture of two homopolymers, amylose and amylopectin, composed of D-glucose units linked together by α (1-4) and α (1-6) bonds which are at the origin of branches in the structure of the molecule.

[0114] These two homopolymers differ in their degree of branching and polymerization. Amylose is slightly branched with short branches and has a molecular mass of 10,000 to 1,000,000 Daltons. The molecule is composed of 600 to 1,000 glucose molecules. Amylopectin is a branched molecule with long branches every 24 to 30 glucose units, via α(1-6) linkages. Its molecular mass ranges from 1,000,000 to 100,000,000 Daltons, and its branching level is approximately 5%. The total chain can contain 10,000 to 100,000 glucose units. The ratio between amylose and amylopectin depends on the botanical source of the starch.

[0115] Starch is stored in reserve organs and tissues in a granular state, that is, in the form of semi-crystalline granules. This semi-crystalline state is primarily due to amylopectin macromolecules.

[0116] In their native state, starch grains have a crystallinity level of 15 to 45% by weight, depending primarily on their botanical origin and the extraction process used. Granular starch, when placed under polarized light, exhibits a characteristic black cross, known as the "Maltese cross," under a microscope. This phenomenon is due to positive birefringence. This is due to the semi-crystalline organization of the granules: the average orientation of the polymer chains is radial. For a more detailed description of granular starch, see Chapter II entitled "Structure and morphology of the starch grain" by S. Perez, in the book "Initiation to macromolecular chemistry and physical chemistry", First Edition, 2000, Volume 13, pages 41 to 86, French Group for the Study and Application of Polymers.

[0117] For the purposes of this application, "damaged starch" means the portion of starch that has been physically broken or fragmented during grain milling. Damaged starch absorbs up to two to three times more water than undamaged starch, which absorbs only about 40% of its own weight. It is also more susceptible to degradation by amylolytic enzymes (both native and added) than undamaged granules that produce fermentable sugars (maltose and glucose) which promote fermentative growth.

[0118] The amount of damaged starch can be analyzed using several protocols, including the international official method ISO 17715:2013 (amperometric method), near-infrared (NIR) reflectance spectroscopy (an indirect method that compares the light absorption of chemical groups in the IR spectrum), or DSC. The international official method ISO 17715:2013 is preferred in this application.

[0119] Preferably, the fraction containing at least 75% by weight of starch that can be obtained by the process according to this application contains more than 80%, preferably more than 90%, even more preferably more than 95% by weight of starch. The starch content of the fraction containing at least 75% by weight of starch that can be obtained by the process according to this application may therefore be 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% as well as all the ranges that can be obtained with two of these values ​​used as upper and lower bounds.

[0120] A sixth object of the present invention is the use of protein concentrate, protein isolate, or mixture thereof, which can be obtained by an extraction process according to the present application, for the manufacture of food or beverage products, in particular plant-based alternatives to milk, animal meat or fish.

[0121] A seventh object of the present invention is the use of the fraction containing at least 75% by weight of starch that can be obtained by the process according to this application, for the manufacture of food or beverage products, in particular plant-based alternatives to milk, animal meat or fish.

[0122] In general, protein concentrate, protein isolate, or a mixture thereof, as well as the final starch-enriched fraction containing at least 75% by weight of starch, obtainable by an extraction process according to the present invention, can be used in food and beverage products. These products may contain from 0.5% to 100% by weight relative to the total dry weight of the food or beverage, for example, from approximately 1% to approximately 80% by weight relative to the total dry weight of the food or beverage. All intermediate values ​​(i.e., 2%, 3%, 4%, ..., 77%, 78%, 79%, 80% by weight relative to the total weight of the food or beverage) can be used, as well as all intermediate ranges based on these quantities. These food and beverage products can be adapted for vegetarian or vegan populations.

[0123] A particularly interesting use of protein concentrate, protein isolate, or mixtures thereof obtainable by an extraction process according to this application concerns their use in beverages. The plant protein compositions of this application can advantageously be used for the manufacture of beverages, in particular milk alternatives, or in other words, milk substitutes.

[0124] In beverages, the protein content can vary widely and can also be a high-protein drink. The protein content can range, for example, from 1 to 12% by dry weight of the beverage, and more specifically from 3 to 10% by dry weight. Beverages can be of all types and include plant-based milk alternatives or milk substitutes, including barista-style milks and coffee creamers.This can also include other ready-to-drink beverages, acidic or not, such as carbonated drinks (including, but not limited to, carbonated soft drinks), non-carbonated drinks (including, but not limited to, non-carbonated soft drinks such as flavored waters, fruit juices, and sweetened or unsweetened tea or coffee-based drinks), alcoholic beverages such as beers or spirits, smoothies, and beverage concentrates (including, but not limited to, liquid concentrates and syrups, as well as non-liquid concentrates, such as freeze-dried and / or powdered preparations or powder mixes). It should be noted that in beverages, flavorings or masking agents are generally used to reduce the pea or broad bean flavor, the bitter aftertaste of the protein, or to flavor the beverage.

[0125] Food products that may be affected by the introduction of protein concentrate, protein isolate, or a mixture thereof, as well as the fraction containing at least 75% by weight of starch that may be obtained by an extraction process according to this application, include bakery products such as bread products (y including, but not limited to, leavened and unleavened breads, sandwich breads, yeast breads and yeast-free breads such as baking soda breads), breads comprising all types of wheat flour, breads comprising all types of flour other than wheat (such as potato, rice, barley, spelt and rye flours), gluten-free breads; mixes for the preparation of said bread products; sweet baked goods (including, but not limited to, rolls, cakes, pies, pastries, waffles, crepes, muffins, pancakes, and biscuits); mixes for the preparation of 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, as well as fillings for cookies, cakes, pastries, confectionery and other products, such as cream fillings); snack bars (including, but not limited to, energy, cereal, nut, and / or fruit bars).

[0126] Due to the presence of damaged starch, the use of the starch-enriched fraction containing at least 75% by weight of starch obtainable by an extraction process according to this application is particularly advantageous because its water absorption capacity is increased compared to a starch-enriched fraction containing at least 75% by weight of starch obtained by a wet extraction process. The starch-enriched fraction containing at least 75% by weight of starch obtainable by an extraction process according to this application is also more susceptible to hydrolysis by amylase (an enzyme capable of breaking down the glucose chains that make up starch). The use of the fraction containing at least 75% by weight of starch obtainable by an extraction process according to this application thus increases the water absorption potential of the flours.The water absorbed by the damaged starch will be released very slowly, which will improve the freshness and shelf life of the bread.

[0127] This can also include set desserts such as custards, flans, and puddings. Another type of dessert can be frozen desserts (including, but not limited to, frozen dairy desserts such as ice cream – including regular ice cream, soft-serve ice cream, and all other types of ice cream – and frozen non-dairy desserts such as non-dairy ice cream, sorbet, and others).

[0128] Other products conventionally prepared from animal milk may also include protein concentrate, protein isolate, or a mixture thereof, as well as the fraction containing at least 75% by weight of starch, obtainable by an extraction process according to this application to form substitutes. These may be acidified and / or fermented products, for example, lactic acid bacteria, or vegan products. or mesophilic. This can include yogurts (including, but not limited to, full-fat, reduced-fat, and fat-free yogurts, which may be free of milk proteins and lactose). The term "yogurts" also includes soft cheeses and fromage frais. It can also include cheese substitutes such as spreadable, processed, cooked and uncooked pressed cheeses, soft cheeses, stretched-curd cheeses, and blue cheeses; it can 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 vegetable butters or crème fraîche.The use of the fraction containing at least 75% by weight of starch, obtainable by an extraction process according to this application, is of particular interest because it is more susceptible to hydrolysis by amylase (an enzyme capable of breaking down the glucose chains that make up starch). The use of the fraction containing at least 75% by weight of starch, obtainable by an extraction process according to this application, thus facilitates and accelerates lactic acid fermentation.

[0129] Other products that may include protein concentrate, protein isolate, or mixtures thereof, which may be obtained by an extraction process according to this application, also include sauces such as salad dressings or sauces based on mayonnaise or ketchup, or syrups.

[0130] Also, protein concentrate, protein isolate, or a mixture thereof, as well as the starch-enriched fraction containing at least 75% starch by weight, obtainable by an extraction process according to this application, may be incorporated into confectionery products (including, but not limited to, gummy candies, soft candies, hard candies, chocolates, caramels, and gums); sweetened and unsweetened breakfast cereals (including, but not limited to, extruded cereals, flaked cereals, and puffed cereals); and cereal coating compositions for the preparation of breakfast cereals. It may also be used in sweetened spreads (including, but not limited to, jellies, jams, nut butters such as peanut butter, spreads, and other spreadable products).

[0131] Protein concentrate, protein isolate, or mixture thereof, as well as starch-enriched fraction containing at least 75% by weight of starch, which may be obtained by an extraction process according to this application, may also be used as a carrier or for flavor encapsulation.

[0132] Other types of food and beverages not mentioned here, but which typically contain one or more proteins, can also be considered within the framework of the present invention. In particular, animal feed (such as pet food) is explicitly considered.

[0133] Protein concentrate, protein isolate, or a mixture thereof, as well as the starch-enriched fraction containing at least 75% starch by weight, obtainable by an extraction process according to this application, may also be used, possibly after texturizing, in meat substitutes such as emulsified sausages or hamburgers, or in fish or seafood substitutes. It may also be used in egg replacement formulations or for the manufacture of protein products such as tofu or tempeh. Textured proteins generally refer to proteins textured by extrusion, i.e., in particular, dry extrusion (also known as Textured Vegetable Protein) or high-moisture extrusion. The extruders may be single-screw, twin-screw, or multi-screw.In twin-screw extrusion, the extrusion can be co-rotating or counter-rotating. Examples of multi-screw extrusion include the planetary extruder and the ring extruder. Other more specialized technologies include shear cell technology, microextrusion, and 3D printing.

[0134] Food products or beverages can be used in specialized nutrition, for example for specific populations such as babies or infants, children, adolescents, adults, the elderly, athletes, and people with illnesses. These can include nutritional meal replacement formulas, complete nutritional drinks (for example, for weight management), or in clinical nutrition (for example, tube feeding or enteral nutrition).

[0135] Protein concentrate, protein isolate, or a mixture thereof, obtainable by an extraction process according to this application, may be used as the sole source of protein, but may also be used in combination with other additional proteins, whether plant or animal. These additional proteins may be hydrolyzed or non-hydrolyzed. Generally, these additional proteins are in the form of concentrates or isolates. Concentrates are distinguished from isolates according to their protein content: concentrates generally have a protein content of 50% to 65% by weight relative to the weight of the concentrate, and isolates have a protein content greater than 80% by weight relative to the weight of the isolate, respectively.The term "plant protein" refers to all proteins derived from cereals, oilseeds, legumes, and tuberous plants, as well as all proteins derived from algae and microalgae or fungi, used alone or in mixtures, chosen from the same family or from different families. By "legume," we mean... Generally, the family of dicotyledonous plants in the order Fabales. Several legumes are important cultivated plants, including soybeans, beans (particularly mung beans), chickpeas, broad beans, peanuts, cultivated lentils, cultivated alfalfa, various clovers, broad beans, carob, licorice, and lupins. The additional legume protein may be selected from these legumes or be a pea or broad bean protein other than that of the invention. In this application, the term "cereals" refers to cultivated plants of the grass family that produce edible grains, for example, wheat, oats, rye, barley, maize, sorghum, or rice. Tubers may include carrots, cassava, konjac, potatoes, Jerusalem artichokes, and sweet potatoes. Oilseed plants are generally plants that produce seeds from which oil is extracted.Oilseed crops can be chosen from sunflower, rapeseed, peanut, sesame, pumpkin, or flax. Animal proteins can be, for example, egg or milk proteins, such as whey protein, casein, or caseinates. The pea or broad bean protein composition of the invention can thus be used in combination with one or more of these proteins or amino acids to improve the nutritional properties of the final product, for example, to improve the PDCAAS (Protein Digestibility Corrected Amnio Acid Score) of the protein or to provide other functionalities.

[0136] Protein concentrate, protein isolate, or a mixture thereof, as well as the starch-enriched fraction containing at least 75% starch by weight, obtainable by an extraction process according to this application, may also be used in the manufacture of pharmaceutical products or in fermentation, for example, for the production of fungal metabolites or cell culture metabolites. The use of the fraction containing at least 75% starch by weight, obtainable by an extraction process according to this application, is of particular interest because it is more susceptible to hydrolysis by amylase (an enzyme capable of breaking down the glucose chains that make up starch). The use of the fraction containing at least 75% starch by weight, obtainable by an extraction process according to this application, thus facilitates and accelerates fermentation processes.

[0137] An eighth object according to the invention consists of the soluble fraction that can be obtained using the extraction process according to the present application.

[0138] The soluble fraction that can be obtained using the extraction process according to this application differs from soluble fractions obtained solely using a single wet process in that its protein content, mainly albumins, and its salt content, mainly potassium, is reduced.

[0139] The invention and its advantages will now be illustrated in the detailed embodiments described in the examples section below. It should be noted that these examples are not exhaustive of the present invention. Examples

[0140] Example 1: Conventional turbo-separation process

[0141] 98.0 kilograms (kg) of pea flour containing 24.0% protein are processed in a Netzsch CFS1000-HD air classifier.

[0142] The process generates 19.6 kg of a light fraction containing 52.0% protein and 78.4 kg of a heavy fraction containing 14.0% protein.

[0143] The protein extraction yield relative to the raw flour (in other words, the amount of protein in the protein concentrate fraction relative to the initial amount of protein in the raw flour) is 44%.

[0144] Example 2: Classical process known as "isoelectric precipitation"

[0145] 96.0 kg of pea flour containing 24.0% protein are suspended in water at 20%. The suspension is then sent to a series of horizontal decanters and centrifugal separators generating 42.3 kg of a starch fraction containing 4% protein.

[0146] The protein-rich supernatant is adjusted to pH 5.0 and then heated for 20 min at 50°C. The treated supernatant is then sent to a centrifugal separator which generates 20.0 kg of a protein isolate fraction containing 85.0% protein.

[0147] The protein extraction yield compared to raw flour is 77%.

[0148] Example 3: Process according to the invention with a mixture of high-protein flour and heavy fraction

[0149] 60.3 kg of pea flour containing 24% protein are sent into a Netzsch CFS1000-HD air classifier to perform dry separation.

[0150] The process generates 12.1 kg of a light fraction called concentrate containing 52.0% by weight of protein and 48.2 kg of a heavy starch-rich fraction containing 14.0% by weight of protein.

[0151] These 48.2 kg of heavy starch-enriched fraction are mixed with 48.3 kg of enriching vegetable flour produced from peas containing 32% protein by weight.

[0152] 96.5 kg of this mixture containing 24.2% by weight of protein are suspended in water at 20%. The suspension is then sent to a series of horizontal decanters and centrifugal separators generating 42.5 kg of a starch-rich fraction containing 4% by weight of protein.

[0153] The protein-rich supernatant is adjusted to pH 5.0 and then heated for 20 min at 50°C. The treated supernatant is then sent to a centrifugal separator which generates 20.0 kg of a protein isolate fraction containing 85.0% protein by weight.

[0154] The protein extraction yield relative to raw flour is 80.3%.

[0155] The protein recovery yield is therefore higher than the processes in examples 1 and 2 (44% and 77% respectively), while not generating a heavy fraction to be valorized.

[0156] This process therefore allows for less costly industrial use, particularly for animal and human food, as well as in the pharmaceutical and nutraceutical industries, with fewer co-products than one of the methods described in Examples 1 and 2 above. Unlike Example 1, this process does not generate a starch-rich fraction contaminated by the presence of protein, and it generates as much usable starch as the process according to the invention in Example 2.

[0157] In an alternative example, still representative of a process according to the invention, 67.8 kg of heavy, starch-rich fraction are mixed with 36.5 kg of enriching vegetable flour produced from peas containing 35% protein by weight. This enriching vegetable flour is considered to have a high protein content. The wet extraction process is then strictly similar to that described in the first part of this example. The process generates 16.9 kg of concentrate containing 52% protein and 20.1 kg of isolate containing 85% protein, with a protein extraction yield relative to the raw flour of 80.1%. 47.3 kg by weight of starch containing 4% protein by weight are also produced.

[0158] Example 4: Process according to the invention with a mixture of low-protein flour and heavy fraction

[0159] 60.3 kg of pea flour containing 24.0% by weight of protein are sent into a Netzsch CFS1000-HD air classifier.

[0160] The process generates 12.0 kg of a light protein-enriched fraction, also called concentrate, containing 52.0% by weight of protein and 48.2 kg of a heavy starch-enriched fraction containing 14.0% by weight of protein.

[0161] These 48.2 kg of heavy fraction are mixed with 64.4 kg of enriching vegetable flour obtained from peas containing 24% protein by weight.

[0162] 112.6 kg of this mixture containing 24.7% by weight of protein are suspended in water at 20%. The suspension is then sent to a series of horizontal decanters and centrifugal separators generating 51.7 kg of a starch fraction containing 4% by weight of protein.

[0163] The protein-rich supernatant is adjusted to pH 5.0 and then heated for 20 min at 50°C. The treated supernatant is then sent to a centrifugal separator which generates 20.0 kg of a protein isolate fraction containing 85.0% protein by weight.

[0164] The protein extraction yield relative to raw flour is 81.0%.

[0165] The protein recovery yield is therefore similar to the process in Example 3, while not generating a heavy, protein-rich fraction requiring further processing. This process generates more usable starch than the process according to the invention in Example 3.

[0166] In an alternative, still representative example of a low-protein flour and heavy fraction mixture, 38.0 kg of heavy fraction are mixed with 70.6 kg of enriching vegetable flour produced from peas containing 24% protein by weight. The wet extraction process is then strictly similar to that described in the first part of this example. This process yields 20.1 kg of isolate at 85% protein by weight and 9.5 kg of concentrate at 52% protein by weight. The protein extraction yield relative to the raw flour is 80.4%. 49.5 kg of starch containing 4% protein by weight are also produced.

[0167] Example 5: Process according to the invention with a mixture of very high protein flour and heavy fraction

[0168] 43.4 kg of pea flour containing 24.0% by weight of protein are sent into a Netzsch CFS1000-HD air classifier.

[0169] The process generates 8.7 kg of a protein-enriched light fraction, also called concentrate, containing 52.0% by weight of protein and 34.7 kg of starch-enriched heavy fraction containing 14.0% by weight of protein.

[0170] These 34.7 kg of heavy fraction are mixed with 34.7 kg of enriching vegetable flour, containing 50% protein by weight.

[0171] 69.4 kg of this mixture are suspended in water at 20%. The suspension is then sent to a series of horizontal decanters and centrifugal separators generating 19.9 kg of a starch fraction containing 4% by weight of protein.

[0172] The protein-rich supernatant is adjusted to pH 5.0 and then heated for 20 min at 50°C. The treated supernatant is then sent to a centrifugal separator which generates 20.0 kg of a protein isolate fraction containing 85.0% protein by weight.

[0173] The protein extraction yield compared to raw vegetable flour is 80.0%.

[0174] The protein recovery yield is therefore similar to the process in Example 2, while not generating a heavy fraction for valorization. This process generates less starch for valorization than the process according to the invention in Example 3.

[0175] Example 6: Synthesis and comparison of the different tests:

[0176] Table 1 below summarizes the different tests:

[0177] The processes according to the invention (Examples 3, 4, and 5) thus eliminate the need to produce a heavy, starch-rich fraction contaminated with more than 4% protein by weight. They also increase the protein extraction yield compared to flour. Choosing the protein content of the enriched flour allows for precise control of the amount of starch produced. This enables a person skilled in the art to produce a protein concentrate, a protein isolate, and a starch-enriched fraction in quantities that can be adjusted.

[0178] Example 6: Comparison of the compositions of soluble fractions obtained using a process according to the invention or outside the invention:

[0179] The soluble fractions obtained in examples 2 (outside the invention) and 3 are recovered in order to analyze the protein content (N 6.25), the total ash content, sodium, potassium and phosphates.

[0180] Table 2 below summarizes the different results:

[0181] The soluble fraction according to the invention contains respectively 24% and 20% less protein and potassium than the soluble fraction obtained with a single wet extraction process.

Claims

Demands

1. A process for producing a protein concentrate and a protein isolate by extracting vegetable protein comprising the following steps: a. Using a vegetable flour having a protein content preferably of 20% to 39%, preferably of 20% to 35% by dry weight relative to the total dry weight of the vegetable flour, b. Separating the vegetable flour from step a) into a protein concentrate, and a starch-enriched fraction using a dry separation process, c. Mixing the starch-enriched fraction obtained in step b) with an enriching vegetable flour, the protein content of which is preferably 20% to 60%, preferably 28% to 36%, even more preferably 30% to 35% by weight, in a mass ratio of starch-enriched fraction / so-called enriching vegetable flour ranging from 70 / 30 to 30 / 70, preferably from 55 / 45 to 35 / 65, d.Separation of the mixture obtained in step c) into a protein isolate, and at least a starch-enriched fraction using a wet separation process.

2. A process according to claim 1 wherein the vegetable flour used in step a) is a legume flour, preferably a pea or broad bean flour or a mixture thereof, preferably a pea flour.

3. A method according to any one of claims 1 to 2 wherein the dry separation method of step b) is selected from a list comprising the turbo-separation method and the electrostatic separation method, preferably the turbo-separation method.

4. A process according to claim 3 wherein the dry separation process of step b) is parameterized to obtain a concentrate containing 40% to 70%, preferably 45% to 60%, preferably 50% to 55% by weight of protein.

5. A process according to any one of claims 1 to 4 wherein the mixture obtained in step c) has a protein content of 20% and 35%, preferably between 20% and 30% by weight.

6. A process according to any one of claims 1 to 5, wherein the wet separation process of step d) is selected from an isoelectric pH protein precipitation process or an ultrafiltration process

7. A process according to claim 6, wherein the wet separation process of step d) is a protein precipitation process at pH isoelectric comprising the following steps: d1) Optionally suspending the mixture obtained in step c) in an aqueous solvent, d2) Extracting the insoluble fractions from the mixture obtained in step c) or from the suspension obtained in step d1) by decantation or filtration, d3) Precipitating the proteins at their isoelectric pH followed by recovering the precipitated proteins by decantation, centrifugation, or filtration, preferably by centrifugation, d4) Optionally rectifying the pH of the proteins precipitated in step d3) to between 5.0 and 9.0, preferably between 6.0 and 8.0, d5) Optionally heat-treating the proteins obtained after step d3) or d4), d6) Optionally shearing the proteins obtained in steps d3), d4), or d5), d7) Optionally homogenizing the proteins obtained in steps d3), d4), d5), or d6), d8) Optionally drying proteins obtained during steps d3), d4), d5), d6) or d7).

8. A process according to claim 7 wherein in step d1) the pH of the mixture obtained in step c) or of the suspension of the mixture obtained in step c) in an aqueous solvent obtained is rectified between 8.0 and 10.0, preferably between 8.5 and 9.

5.

9. A process according to claim 7 or 8 wherein the precipitation of proteins at their isoelectric pH of step d3) is carried out at a pH of 4 to 6, preferably 5, followed by heating to a temperature of 50°C 7.2°C to 60°C 7.2°C, preferably 52°C 7.2°C to 58°C 7.2°C.

10. Starch-enriched fraction containing at least 75% by weight of starch capable of being obtained by the process according to any one of claims 1 to 9.

11. Fraction according to claim 10 containing more than 80%, preferably more than 90%, even more preferably more than 95% by weight of starch.

12. A powder mixture consisting of a starch-enriched fraction obtained by means of a dry separation process which can be obtained by the process according to any one of claims 1 to 9 and an enriching vegetable flour having a protein content of 20% to 60%, preferably 28% to 36%, even more preferably 30% to 35% by weight, in a mass ratio of starch-enriched fraction / vegetable flour ranging from 70 / 30 to 30 / 70, preferably from 55 / 45 to 35 / 65.

13. Soluble fraction obtainable by the process according to any one of claims 1 to 9.

14. Use of the starch-enriched fraction according to claim 10 or 11, for the manufacture of food or beverage products, in particular plant-based alternatives to milk, animal meat or fish.

15. Use of the protein concentrate, protein isolate, or mixture thereof, which can be obtained by a process according to any one of claims 1 to 9, for the manufacture of food or beverage products, in particular plant-based alternatives to milk, animal meat or fish.

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