Process for obtaining a texturising protein extract from microalgae

Abstract

The present invention relates to a process for obtaining a protein extract from microalgae. The protein extract obtained has a texturising ability to form a protein network, pure or in a mixture with another protein source, allowing the formation of food gels, extruded protein material or cooked cereal products. The protein extract obtained also has properties of coagulation, pure or in a mixture with another protein source, allowing the formation of food products that require protein coagulation without the formation of a protein network, such as hot emulsions or desserts ("custards"). The invention also relates to the use of the protein extract for replacing the texturising functionalities provided by animal proteins (egg proteins, dairy proteins, red or white meat proteins) within these products, and therefore for replacing sources of animal protein within these products.

Description

Description Title of the invention: Process for obtaining a texturizing protein extract from microalgae.

[0001] This patent application relates to a process for obtaining a protein extract from microalgae. The resulting protein extract exhibits a texturizing capacity for protein network formation, either pure or in mixture with another protein source, enabling the formation of food gels, extruded protein materials, or cooked cereal products. The protein extract also exhibits coagulation properties, either pure or in mixture with another protein source, enabling the formation of food products requiring protein coagulation without protein network formation, such as hot emulsions or custards.The invention also relates to the use of the protein extract to substitute the texturizing functionalities provided by animal proteins (egg proteins, dairy proteins, red or white meat proteins) in these products, and thus to replace the sources of animal proteins in these products. Previous art

[0002] There is a variety of chemical, physical or enzymatic processes known to the skilled person to obtain a protein extract, isolate or concentrate (isolate for a protein content greater than or equal to 80%, concentrate for a protein content between 50% and 80%) from a plant or microbial raw material.

[0003] These processes are combined in a specific sequence with the aim of successively carrying out the operations of grinding the raw material, extracting the protein material, concentrating and / or purifying the extracted protein material.

[0004] The objective of combining extraction, concentration and / or purification steps is to obtain a protein extract with sufficient protein content (in the case of an isolate, greater than 50%) as well as targeted functional and / or nutritional properties.

[0005] Functional properties correspond to the ability of the protein extract to be transformed into a food matrix of which it forms all or part, and to impart specific organoleptic properties to this matrix.

[0006] The organoleptic properties provided by the protein extract are properties of texture, appearance (color, visual texture, etc.), smell, taste and aftertaste.

[0007] The nutritional properties provided by the protein extract may include the essential and non-essential amino acid profile, protein digestibility, and protein digestion kinetics.

[0008] The functional properties of the protein extract depend on the physicochemical properties of the extract after the extraction, concentration and / or purification steps.

[0009] The determining physicochemical properties of the protein extract include: solubility of the protein extract, protein fraction composition, size of protein fractions, tertiary and quaternary structures of the protein fractions, amino acid sequence of the protein fractions.

[0010] The physicochemical properties of the protein extract are impacted by the conditions under which the chemical, physical or enzymatic extraction, concentration and / or purification processes are carried out, as well as the sequence in which these processes are linked.

[0011] Microalgae are a particularly interesting source of protein due to their high protein content, generally between 40 and 60% in dry matter, and their high content of essential amino acids (no deficiency unlike with classic plant protein sources in food).

[0012] Furthermore, these raw materials have a significantly higher productivity per square meter than plant crops, on the order of 10 to 50 g / m². 2 / day for example for the microalga Chlorella vulgaris versus 50 to 500 g / m 2for a single harvest season of legume or wheat crops (i.e., an equivalent productivity of 0.15 to 5 g / m²) 2 / day). The absence of seasonality in microalgae cultivation under controlled systems gives this raw material a significant advantage and strong potential in terms of security of supply.

[0013] Finally, unlike conventional plant-based agricultural systems, microalgae cultivation does not require the use of insecticides or pesticides. It also requires approximately eight times less nitrogen than conventional wheat cultivation (46 g / kg of raw material produced, compared to 360 g / kg).

[0014] However, there are few methods for obtaining protein extracts from microalgae.

[0015] The WO 2020 / 261245 document from NOBLEGEN INC. describes process conditions enabling the production of Euglena sp. flour, concentrate and isolate, as well as food product compositions containing these extracts.

[0016] However, no capacity of the flour, concentrate or The isolate is intended to provide sufficient texturizing capacity to allow it to replace animal proteins (egg proteins, dairy proteins, or red or white meat proteins) in the targeted food products. No protein network formation or protein coagulation capacity is claimed, either on its own or in mixture with another protein source.

[0017] Indeed, the flours, concentrates or isolates from the process are used as a partial substitute for the structuring ingredient they are supposed to replace, for example up to 30% of wheat flour in pasta, or 55% of pea flour in protein extracts.

[0018] Document EP 3622828 from CORBION BIOTECH INC. presents Chlorella sp. compositions in the form of flakes, powders, and flours. These products are characterized by a high content of unsaturated lipids, fiber, and protein, and can be used as a partial replacement for egg or fat in food products. Examples of food product compositions including these products are provided.

[0019] Similarly, the document does not disclose any capacity of Chlorella sp. compositions to provide sufficient texturizing capacity to completely replace animal proteins (egg proteins, dairy proteins, or red or white meat proteins) in the targeted food products. No protein network formation or protein coagulation capacity is claimed, either on its own or in mixture with another protein source.

[0020] Outside the field of microalgae, among existing protein extraction processes, US document 2017 / 0238590 from HAMPTON CREEK, INC. presents process conditions allowing the obtaining of mung bean protein extracts varying in composition (protein content, composition in protein fractions, size, structure and amino acid sequence of the fractions, lipid content, composition in lipid fractions, in particular).

[0021] The document discloses a texturizing capacity of the protein extracts produced and more specifically a capacity of these extracts to provide the main functional properties provided by the egg (solubility, gelation, emulsification, foaming) and therefore to replace the egg in certain food products.

[0022] The document demonstrates how these functional properties are imparted by protein extracts in the following food products: vegan scrambled egg substitutes, soft egg-free cakes, pasta, vegan spreadable cheese, vegan milk substitute, vegan butter substitute, and vegan meat substitute. It presents formulas and preparation methods for all of these products.

[0023] However, document US 2017 / 0238590 is limited to obtaining extracts protein from mung beans and does not mention the possibility of using microalgae as a raw material.

[0024] Furthermore, it is not possible for a person skilled in the art to obtain a protein extract from microalgae exhibiting a protein network-forming texturizing capacity, either pure or in mixture with another protein source, capable of replacing animal proteins (egg proteins, milk proteins, red or white meat proteins) in food gels, coagulated sauces and desserts, extruded protein material, or cooked cereal products, using the process steps indicated by HAMPTON CREEK, INC. US 2017 / 0238590, whether intentionally or randomly.

[0025] Indeed, this is due to the fact that the raw materials mung bean (Vigna radiata) and microalgae differ significantly in their initial composition and histological structure. These elements directly determine the steps of the chemical, physical, or enzymatic processes known to those skilled in the art that must be implemented, as well as their sequence, to enable the grinding of the raw material, followed by the extraction, concentration, and / or purification of the protein extract.

[0026] Indeed, Vigna radiata seeds are composed of approximately 15 to 30% protein, 55 to 65% carbohydrates (including 3 to 6% fiber), and 1 to 7% lipids. Histologically, the Vigna radiata seed consists of a predominantly fibrous, easily detachable seed coat (the "testa") surrounding two cotyledons. The cotyledons are composed of plant cells containing starch granules surrounded by a protein matrix.

[0027] Microalgae, on the other hand, are unicellular or multicellular organisms with higher protein and lipid content, ranging from 30-60% to 2-50% on a dry basis, respectively. Furthermore, the presence of a rigid outer cell wall rich in cellulose, hemicellulose, and / or lignin necessitates specific chemical, physical, or enzymatic steps at the beginning of the protein extract extraction process.

[0028] Thus, there remains a need for a process enabling the obtaining of a protein extract from microalgae, which is adapted to the composition and histological structure of these raw materials, and which allows the obtaining of protein extract(s) capable of substituting the texturizing functionalities provided by animal proteins (egg proteins, dairy proteins, red or white meat proteins) in food products, particularly food gels, coagulated sauces and desserts, extruded protein materials and cooked cereal products.

[0029] The applicant resolved this technical problem by proposing conditions of A process specifically adapted for obtaining protein extracts from microalgae. The resulting protein extracts exhibit a texturizing capacity for protein network formation, either pure or mixed with another protein source, enabling the formation of food gels, extruded protein materials, or cooked cereal products. They also possess a texturizing capacity for coagulation, either pure or mixed with another protein source, allowing the formation of coagulated sauces and desserts. Therefore, the protein extracts produced, alone or mixed with another protein source, can replace the texturizing properties provided by animal proteins (egg proteins, dairy proteins, red or white meat proteins) in these products, and thus replace animal protein sources altogether.The protein extracts produced are in the form of liquid extracts or powders with a water content of less than or equal to 5% w / w (protein isolates). Description of the invention

[0030] Thus, according to a first aspect, the invention relates to a process for obtaining protein extract(s) with texturizing capacity from microalgae.

[0031] According to one embodiment, said process for obtaining protein extract(s) with texturizing capacity from microalgae according to the invention comprises at least the successive steps of: a. Preparation of an aqueous suspension of microalgae with a concentration of between 1% and 20% w / w of microalgae, and a pH of between 7 and 12, at a temperature of between 4 and 25°C; c. Rupture of the rigid outer cell wall of the microalgae, preferably at a temperature of between 4 and 25°C by treatment of the aqueous suspension by application of ultrasound, by high-pressure homogenization or using a ball mill; d. Extraction of the aqueous protein phase from the liquid extract of microalgae with ruptured cell walls, preferably at a temperature of between 4 and 25°C by centrifugation or three-phase separation by solvents; o.Extraction of a protein concentrate from the aqueous protein phase, preferably by isoelectric precipitation, precipitation by variation of ionic strength, tangential ultrafiltration, cryo-precipitation or by a combination of these techniques; p. Dilution of the protein concentrate in an aqueous phase in a ratio of between 1:1 and 1:5 v / v, preferably under the same conditions of pH, ionic strength and temperature used in step o.

[0032] The numbers for these steps are not in direct sequence, for clarity. in defining the optional steps that will follow in the development of this description.

[0033] Preferably, the microalgae used in step a. are: i. in dry form, with a water content between 1 and 10% (w / w); or ii. in frozen form, with a water content between 10 and 90% (w / w);

[0034] Preferably, the microalgae used in step a. have a protein content greater than 45% (w / w) on a dry basis.

[0035] Preferably, in step a., the pH of the aqueous microalgae suspension is adjusted by the use of food-grade sodium hydroxide (NaOH, "caustic soda") with a concentration between 0.5 and 5 mol / L.

[0036] Preferably, cell wall rupture in step c is achieved by application of ultrasound, high pressure homogenization (HPH) or use of a ball mill.

[0037] Preferably, step c, which involves breaking down the rigid outer cell wall of the microalgae in the homogenized suspension obtained in step a or b, is carried out: i. Either by applying ultrasound at a frequency between 10 and 40 kHz, at an amplitude between 30 and 60%, at a power between 50 and 1000 W, for a duration between 5 and 60 minutes, divided or not into 30-second intervals; or ii. Or by high-pressure homogenization (HPH), from 1 to 5 cycles at a pressure between 800 bar and 3000 bar; or iii. Or by using a ball mill, with balls of diameter 0.4 to 2 mm, filling the extraction chamber between 60 and 90%, at a rotation speed of 1000 to 4000 rpm for 5 to 90 minutes.

[0038] Preferably, the extraction of the aqueous protein phase from the liquid extract of broken-walled microalgae obtained in step d. is carried out by centrifugation or three-phase solvent partitioning.

[0039] Preferably, step d, extraction of the aqueous protein phase from the liquid extract obtained in step c, is carried out: i. Either by centrifugation at a speed of between 5000 and 15000 g for 10 to 30 minutes; or ii. Or by three-phase solvent partitioning by mixing the liquid extract obtained in step c with ammonium sulfate in a ratio of between 1:3 and 5:1 (w / v) and t- butanol in a ratio between 1:2 and 10:1 (v / v), for a duration of between 5 and 30 minutes. After decantation, the intermediate phase containing the proteins is recovered.

[0040] Preferably, step o. of protein concentrate extraction is carried out by isoelectric precipitation, precipitation by ionic strength variation, tangential ultrafiltration, cryoprecipitation or by a combination of these techniques;

[0041] Preferably, in step o., the extraction of a protein concentrate from the aqueous protein phase obtained in step d., e., f., i., l. or m. is carried out: i. Either by isoelectric precipitation by mixing the aqueous protein phase obtained in step d., e., f., i., l. or m. with citric acid approved for food contact so as to achieve a pH between 3.5 and 8, more preferably between 3.5 and 5.5; or ii. Or by precipitation by variation of ionic strength by mixing the aqueous protein phase obtained in step d., e., f., i., l. or m. with food salt so as to achieve an ionic strength between 0.15 and 5M; or iii.Either by tangential ultrafiltration using a tangential ultrafiltration unit equipped with a polyethersulfone membrane with a pore size between 50 and 500 microns, with a transmembrane pressure between 0.2 and 3 bars, and a number of passes within the unit between 1 and 5; or iv. Or by cryoprecipitation by cooling the aqueous protein phase obtained in steps d, e, f, i, l or m to a temperature between 0 and 3 °C; or v. Or by a combination of these techniques;.

[0042] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises between step a. and step c. a step b. of homogenizing the aqueous suspension of microalgae obtained in step a. at a temperature between 4 and 25°C.

[0043] Preferably, in step b., the homogenization of the aqueous microalgae suspension is carried out using a rotor-stator homogenizer or disperser (for example a VORTEX agitator or an ULTRA-TURRAX disperser from the manufacturer IK A-Werke, Germany) at a speed between 500 and 5000 rpm, for a period of 10 to 60 seconds.

[0044] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises, after step d, a step e of diluting the protein extract obtained in step d in a aqueous phase in a ratio between 1:1 and 1:10 v / v of the initial volume of the protein extract, at a pH between 7 and 12 and at a temperature between 4 and 25°C.

[0045] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises, after step d. or step e. if present, a step f. of extraction of the carbohydrate fraction contained in the aqueous protein phase obtained in step d. or e. by solid / liquid separation or centrifugation, at a temperature between 4 and 25°C.

[0046] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises the steps of: g. mixing the carbohydrate fraction obtained in step f. in an aqueous phase in a ratio of between 1:2 and 1:10 v / v, at a pH of between 7 and 12 at a temperature of between 4 and 25°C; h. centrifuging and / or filtering the aqueous suspension of carbohydrate fraction obtained in step g., at a temperature of between 4 and 25°C; i. mixing the residual aqueous protein extract obtained in step h. with the residual aqueous protein phase obtained in step d. or f.

[0047] Preferably, during steps e. and g., the pH is adjusted by the use of food-grade sodium hydroxide (NaOH, "caustic soda") with a concentration between 0.5 and 5 mol / L;

[0048] Preferably, in step f, the extraction of the carbohydrate fraction from the aqueous protein phase obtained in step d or e is carried out: i. Either by solid / liquid separation using a decanter centrifuge or disc separator at a speed between 1000 and 15000g; or ii. Or by centrifugation at a speed between 5000 and 15000g for 10 to 30 minutes;

[0049] Preferably, in step h, the aqueous suspension of carbohydrate fraction obtained in step g is treated: i. Either by centrifugation at a speed of between 5000 and 15000g for 10 to 30 minutes; or ii. Or by membrane filtration with a pore size of between 50 and 500 microns; or iii. Or by centrifugation at a speed of between 5000 and 15000g for 10 to 30 minutes followed by membrane filtration with a pore size of between 50 and 500 microns.

[0050] According to one embodiment, said process for obtaining extract(s) protein(s) from microalgae according to the invention comprises the steps of: j. mixing the aqueous protein phase obtained in step d., e., f. or i. with transglutaminase at a concentration of between 5 and 50 units / g of protein, at a temperature of between 25 and 55°C, for 10 to 120 minutes; k. inactivation of the transglutaminase in the mixture obtained in step j. by heating at a temperature of between 60 and 80°C; l. centrifugation of the mixture obtained in step k. at a temperature of between 4 and 55°C.

[0051] Preferably, in step 1, the mixture obtained in step k is centrifuged at a speed between 1000 and 5000g for 5 to 30 minutes.

[0052] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises the steps of: m. purification of the aqueous protein phase obtained in step d., e., f., i. or 1. by dialysis or adsorption on carbon particles, at a temperature between 4 and 25°C; n. centrifugation of the purified aqueous protein phase obtained in step m. at a temperature between 25 and 55°C.

[0053] Preferably, in step m., the purification of the aqueous protein phase obtained in step d., e., f., i. or 1. is carried out: i. Either by dialysis (diafiltration) against osmosis, distilled, demineralized or milli-Q water at a pH between 7 and 12 using a membrane with a pore size between 10 and 100 microns, for 30 minutes to 12 h; or ii. Or by adsorption on carbon particles by mixing the aqueous protein phase obtained in step d., e., f., i. or 1. with food-grade carbon particles, with a size between 100 microns and 5 mm in diameter, in a ratio between 1:1 and 1:20 (w / w), for 4 to 30 minutes;

[0054] Preferably, in step n., the mixture obtained in step m. is centrifuged at a speed between 5000 and 15000g for 10 to 30 minutes.

[0055] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises, after step p., a step q. of centrifugation of the aqueous protein phase obtained in step p., preferably under the temperature conditions used in step o.

[0056] Preferably, in step q, the aqueous protein phase obtained in step p is centrifuged at a speed between 5000 and 15000 g / s for 10 to 30 minutes, under the temperature conditions used in step o. ;

[0057] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises a step r. of filtration of the aqueous protein phase obtained in step p. or q. by ultrafiltration, at a temperature between 4 and 25°C.

[0058] Preferably, in step r, the filtration of the aqueous protein phase obtained in step p or q is carried out by tangential ultrafiltration using a tangential ultrafiltration unit equipped with a polyethersulfone membrane with a pore size between 50 and 500 microns, with a transmembrane pressure between 0.2 and 3 bars, and a number of passes within the unit between 1 and 5.

[0059] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises a step s of diluting the aqueous protein phase obtained in step p., q. or r. with water at a pH between 7 and 12 and at a temperature between 4 and 25°C, so as to obtain a water content between 70 and 95% v / v.

[0060] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises a step t. of flash-pasteurization of the aqueous protein phase obtained in step p., q., r. or s. at a temperature between 70 and 75°C for 15 to 25 seconds, so as to obtain a liquid protein extract with a water content between 65 and 95% v / v.

[0061] Preferably, in step u., the drying of the aqueous protein phase obtained in step p., q., r., s. or t. is carried out by spraying, with an inlet temperature of the sprayer between 150 and 200°C, and an outlet temperature of the sprayer between 50 and 150°C;

[0062] According to one embodiment, said process for obtaining protein extract(s) from microalgae according to the invention comprises a step u. of drying the aqueous protein phase obtained in step p., q., r., s. or t. by freeze-drying, spray drying, low temperature drying or pressure drying, preferably by spray drying or freeze-drying, so as to obtain a protein extract in the form of a powder with a water content less than or equal to 5% w / w.

[0063] The term "microalgae" in the present invention refers to eukaryotic microalgae, which are characterized by a nucleus, including, for example, chlorophytes, rhodophytes, haptophytes, bacillariophytes, eustigmatophytes, euglenophytes, thraustochytriaceae, or dinophytes, said eukaryotic microalgae being commonly referred to as "microalgae," and prokaryotic microalgae, which do not possess a nucleus, including cyanophytes, ci- later specifically named "cyanobacteria".

[0064] Preferably according to the invention, eukaryotic microalgae are selected from chlorophytes, preferably from Chlorella, Auxenochlorella, Dunaliella, Tetraselmis, Haematococcus, Scenedesmus; eustigmatophytes, preferably Nannochloropsis; euglenophytes, preferably Euglena; rhodophytes, preferably Porphyridium; bacillariophyceae, preferably Phaeodactylum and Odontella, and thraustochytriaceae, preferably Schizochytrium.

[0065] Preferably, cyanobacteria are chosen from spirulina (Arthrospira platensis, Limnospira platensis or Spirulina maxima) and AFA (Aphanizomenon Floes-aquae).

[0066] Even more preferably, said microalgae is chosen from Chlorella, Chlorella sp., Chlorella vulgaris, Chlorella protothecoides, Chlorella sorokiniana, Chlorella pyrenoidosa, Euglena sp., Euglena gracilis, Arthrospira platensis, Limnospira platensis, Spirulina maxima, Tretraselmis sp., Tretraselmis chuii., Nannochloropsis sp., Dunaliella sp., Scenedesmus sp., Aphanizomenon sp., Haematococcus sp.

[0067] Even more preferably, where said microalgae are chosen from Chlorella vulgaris and Tretraselmis chuii.

[0068] According to one embodiment of the invention, said process for obtaining protein extract(s) from microalgae according to the invention therefore comprises the following steps: a. Preparation of an aqueous suspension of microalgae with a concentration of between 1 and 20% w / w of microalgae, and a pH of between 7 and 12, at a temperature of between 4 and 25°C; b. Optionally, homogenization of the aqueous suspension of microalgae obtained in step a. at a temperature of between 4 and 25°C; c. Rupture of the rigid outer cell wall of the microalgae, preferably at a temperature of between 4 and 25°C by treating the suspension obtained in step a. or b. by application of ultrasound, by high-pressure homogenization (HPH) or using a ball mill; d. Extraction of the aqueous protein phase from the liquid extract of microalgae with ruptured cell walls obtained in step c., preferably at a temperature between 4 and 25°C by centrifugation or three-phase separation by solvents; e. Optionally, dilution of the protein extract obtained in step d. in an aqueous phase at a ratio between 1:1 and 1:10 v / v of the initial volume of the protein extract, at a pH between 7 and 12 and at a temperature between 4 and 25°C; f. Optionally, extraction of the carbohydrate fraction contained in the phase. aqueous protein obtained in step d. or e. by solid / liquid separation or centrifugation, at a temperature between 4 and 25°C; g. Optionally, mixing the carbohydrate fraction obtained in step f. in an aqueous phase in a ratio between 1:2 and 1:10 v / v, at a pH between 7 and 12 at a temperature between 4 and 25°C; h. Optionally, centrifugation and / or filtration of the aqueous suspension of carbohydrate fraction obtained in step g., at a temperature between 4 and 25°C; i. Optionally, mixing the residual aqueous protein extract obtained in step h. with the residual aqueous protein phase obtained in step d. or f.; j. Optionally, mixing the aqueous protein phase obtained in steps d., e., f. or i. with transglutaminase at a concentration of between 5 and 50 units / g of protein, at a temperature between 25 and 55°C, for 10 to 120 minutes; k.Optionally, inactivation of transglutaminase in the mixture obtained in step j by heating at a temperature between 60 and 80°C; l. Optionally, centrifugation of the mixture obtained in step k at a temperature between 4 and 55°C; m. Optionally, purification of the aqueous protein phase obtained in steps d, e, f, i, or 1 by dialysis (diafiltration) or adsorption onto charcoal particles at a temperature between 4 and 25°C; n. Optionally, centrifugation of the purified aqueous protein phase obtained in step m at a temperature between 25 and 55°C; o. Extraction of a protein concentrate from the aqueous protein phase obtained in steps d, e, f, i, 1, m. or n., preferably by isoelectric precipitation, precipitation by variation of ionic strength, tangential ultrafiltration, cryoprecipitation or by a combination of these techniques; p. Dilution of the protein concentrate obtained in step o.in an aqueous phase at a ratio of 1:1 to 1:5 v / v, preferably under the same pH, ionic strength and temperature conditions used in step o; q. Optionally, centrifugation of the aqueous protein phase obtained in step p under the temperature conditions used in step o; r. Optionally, filtration of the aqueous protein phase obtained in step p or q by ultrafiltration at a temperature between 4 and 25°C; s. Optionally, dilution of the aqueous protein phase obtained in step p, q or r with water at a pH between 7 and 12 and a temperature between 4 and 25°C, so as to obtain a water content between 70 and 95% v / v; t. Optionally, flash-pasteurization of the aqueous protein phase obtained. at step p., q., r. or s. at a temperature between 70 and 75°C for 15 to 25 seconds, so as to obtain a liquid protein extract with a water content between 65 and 95% v / v; u. Optionally, drying of the aqueous protein phase obtained at step p., q., r., s. or t. by freeze-drying, spray-drying, low-temperature drying or pressure drying, preferably by spray-drying or freeze-drying, so as to obtain a protein extract in powder form with a water content less than or equal to 5% w / w.

[0069] The term "carbohydrate fraction" refers to the portion comprising fibers and starch.

[0070] The term "aqueous suspension of microalgae" refers to a mixture of microalgae and water.

[0071] By "on a dry basis" is meant that the microalga does not contain water, and is therefore in solid form, for example as a powder.

[0072] By "rupture of the rigid outer cell wall" is meant the lysis of the cell membrane of microalgae in order to obtain all of its membrane and intracellular contents.

[0073] By "aqueous protein phase" is meant an aqueous mixture comprising proteins from the microalga.

[0074] For the purposes of this invention, "high pressure homogenization" means that the mixing is carried out in a device capable, by applying significant pressure, of creating microscopic particles that allow substances to be mixed.

[0075] According to the invention, "homogenization" means the process of harmonizing the different elements of a mixture.

[0076] "Filtration" is understood to be a separation process that allows the components of a mixture with a liquid phase and a solid phase to be separated through a porous medium whose size is defined according to the elements to be separated.

[0077] For the purposes of this invention, "centrifugation" refers to a technique that uses centrifugal force to separate the phases of a mixture. The mixture is subjected to a very rapid rotation, and the heavier particles are pushed towards the walls of the container by the centrifugal force, while the lighter particles and liquids remain on the surface (supernatant).

[0078] For the purposes of this invention, "ball milling" refers to the use of a device consisting of a horizontal drum rotated by a motor. The drum is partially filled with the product to be ground, to which the grinding elements (e.g., metal balls) are added.

[0079] The term "three-phase solvent separation" refers to this method of Separation, purification, and concentration are achieved by the sequential addition of a sufficient amount of salt (typically ammonium sulfate) and an organic solvent (primarily β-butanol / β-butanol). An organic solvent such as β-butanol is added to form three-phase layers and to remove low molecular weight compounds such as lipids, phenolic compounds, and certain detergents. After processing the crude extract and decantation, the mixture separates into three distinct phases: the upper solvent (β-butanol) contains nonpolar compounds, which are separated from the lower aqueous phase (containing polar compounds) by a protein precipitate.

[0080] By "protein concentrate" is meant a solid phase comprising microalgae proteins.

[0081] Isoelectric precipitation refers to the process of adjusting the pH of the solution to the isoelectric point of the protein to be extracted, thereby enabling its selective purification. Indeed, at the isoelectric point, a protein tends to become less soluble.

[0082] "Precipitation by variation of ionic strength" is understood to mean increasing the concentration of the mixture in salt, so as to decrease the solubility in water of certain substances such as proteins.

[0083] Tangential flow ultrafiltration refers to a filtration process designed to separate particles in a liquid based on their size. In this type of filtration, the liquid flows parallel to the filter, and it is the fluid pressure that forces the liquid through the filter. As a result, relatively small particles pass through the filter, while larger particles continue through the flow.

[0084] "Cryoprecipitation" refers to the phenomenon of slowly thawing the solution at low temperatures, followed by centrifugation and separation of the supernatant.

[0085] By "residual protein aqueous extract" is meant the proteins remaining in the aqueous extract despite the previous steps.

[0086] "Solid / liquid separation" consists of separating two phases, solid and liquid, of a suspension.

[0087] "Membrane filtration" refers to this physical separation process that takes place in the liquid phase. The aim is to purify, fractionate, or concentrate dissolved or suspended species in a solvent by passing them through a membrane.

[0088] Transglutaminase is an enzyme that catalyzes the formation of covalent bonds between free amine groups, such as those of lysine residues, and the gamma-carboxamide group of glutamine residues. The bonds formed by transglutaminase exhibit high resistance to proteolysis. Transglutaminase forms protein polymers, generally insoluble.

[0089] Transglutaminase inactivation is understood to mean applying heat above 50°C, in order to denature this enzyme and stop its activity.

[0090] By "dialysis" or "diafiltration" is understood this dilution process which involves the elimination or separation of certain components of a liquid containing soluble molecules and under certain filterable conditions (e.g. salts, sugars, small proteins, solvents) in a solution (aqueous in general), based on sorting the molecules according to their size using more or less nanometric permeable filters.

[0091] "Adsorption onto carbon particles" refers to the process of attaching proteins from a liquid phase to the surface of carbon particles. Activated carbon has the greatest physical adsorption strength and the largest adsorption volume of any known natural or synthetic material.

[0092] "Flash pasteurization" or rapid pasteurization is understood to mean passing the mixture through a continuous and controlled flow, and subjecting it to a temperature between 71.5 °C and 74 °C for 15 to 30 seconds, followed by rapid cooling.

[0093] "Spray drying" refers to the process of dehydrating a liquid mixture into a powder form in a stream of hot air.

[0094] Freeze-drying refers to the process of freezing food and then dehydrating it to remove water. A freeze-drying cycle comprises three main phases: freezing, primary drying, and secondary drying.

[0095] By "low temperature drying" we mean drying the product to obtain a solid result, without using excessively high temperatures, for example below 45°C.

[0096] By "pressure drying" is understood that drying is achieved by applying pressure to the liquid, for example by passing it through an orifice or a narrow conduit.

[0097] By "protein network" is meant the presence of chemical interactions of a covalent, hydrophobic, ionic, hydrogen, Van der Waals or steric nature between peptide chains belonging to proteins originating from the same raw material ("pure network") or originating from different raw materials ("mixed network") allowing to form a 3D mesh at the molecular scale capable of structuring the food matrix which contains it, and consequently of giving it textural properties.

[0098] "Gelification" refers to the formation of a food gel, corresponding to the formation of a protein network that traps molecules of water within an aqueous phase that contains it.

[0099] "Coagulation" refers to the denaturation, unfolding and aggregation of peptide chains belonging to proteins originating from the same raw material ("pure coagulation") or from different raw materials ("mixed coagulation") to provide viscosity to the food matrix.

[0100] By "texturizing capacity" is meant the ability of the protein extract to provide a variable level of texturization to the food matrix of which it forms all or part, ranging from a minimal increase in viscosity for use of the protein extract in low concentration to the formation of a viscoelastic protein network capable of providing firmness, rigidity and elasticity to the food matrix for use in higher concentration.

[0101] The term "protein extract" refers to a product obtained in aqueous or solid form, for example as a powder, comprising proteins derived from microalgae. These proteins are between 5 and 600 kDa in size.

[0102] By "protein isolate" is meant an extract composed of proteins at more than 80% of its total mass on a wet basis.

[0103] By “protein concentrate” is meant an extract composed of proteins at a level of 50 to 80% of its total mass on a wet basis.

[0104] The term “flour” is understood to mean a dry and ground form of a raw material including all or part of the composition of that raw material, and with a particle size of less than 300 microns.

[0105] By "histological structure" is meant the organization of plant tissues in relation to each other within the plant raw material, whether it comes from plants or algae.

[0106] According to a second aspect, the invention relates to a protein extract with texturizing capacity as obtained by the process according to the invention.

[0107] Preferably, said protein extract is in the form of a liquid extract or a powder with a water content less than or equal to 5% w / w (protein isolate).

[0108] According to a third aspect, the invention relates to the use of the protein extract obtained according to the process according to the invention as a substitute for texturizing animal proteins in food products.

[0109] The extract replaces the texturizing functionalities provided by animal proteins, preferably egg proteins, dairy proteins, red or white meat proteins in food products.

[0110] According to one embodiment, the protein extract obtained by the process according to the invention is introduced into the composition of a food product at a level of between 1% and 80% of the total composition of the food product. [OR I] Preferably, said food products are chosen from among the gels food products, coagulated sauces and desserts, extruded protein materials and cooked cereal products.

[0112] In the description and the following examples, unless otherwise stated, percentages are percentages by weight, and value ranges expressed as "between ... and ..." include the specified lower and upper bounds. The examples below are provided for illustrative purposes only and are not intended to limit the scope of the invention.

[0113] In the description and in the examples, the term v / v means a "volume to volume" ratio, and signifies a percentage or quantity in volume relative to a total volume.

[0114] In the description and examples, the term w / w means a "weight-for-weight" ratio, and signifies a percentage or quantity by weight relative to a total weight. Examples

[0115] Example 1: Process for producing protein isolate from Tetraselmis chuii.

[0116] The process according to the invention is used to produce a protein isolate of Tetraselmis chuii.

[0117] The process steps and conditions used to produce the protein isolate are presented in Table 1. The step numbers correspond to the numbers used in the claims and in the description.

[0118] [Table 1]

[0119] Example 2: Process for producing a liquid protein extract of Chlorella vulgaris.

[0120] The process according to the invention is used to produce a liquid protein extract of Chlorella vulgaris.

[0121] The process steps and conditions used to produce the liquid protein extract are presented in Table 2. The step numbers correspond to the numbers used in the claims and in the description.

[0122] [Table 2] _

[0123] Example 3: Comparison of the texturizing capacity of coagulation and gelling of the liquid protein extract of Chlorella vulgaris prepared according to example 2 and of egg, soy and pea proteins.

[0124] The process according to the invention is used to produce a liquid protein extract of Chlorella vulgaris according to example 2.

[0125] The ability of the protein extract to coagulate and then form a protein network, pure or mixed with another protein source in the form of an aqueous food gel, is tested and compared to the ability of egg and pea proteins.

[0126] Two solutions are prepared for each type of protein studied, in water at pH 8 at 25°C: one with 5% (w / v) of protein to test the coagulation capacity, the other with 10% (w / v) of protein to test the gelation capacity.

[0127] The samples tested in this test are presented in Table 3.

[0128] [Table 3]

[0129] Each sample solution presented in Table 3 is prepared in triplicate.

[0130] The ability to coagulate or gel of each sample is measured by monitoring the dissipative (or viscous) modulus G” and the conservative (or elastic) modulus G' during a temperature variation using a rheometer equipped with a coaxial cylinder Couette cell and a Peltier system for temperature regulation.

[0131] The measurement protocol is as follows: temperature rise from 25 to 95°C at a rate of 2.5°C / min (frequency 1Hz, deformation 1%) followed by a plateau at 85°C of 15 minutes then a temperature descent from 95°C to 25°C at a rate of 2.5°C / min.

[0132] To assess coagulation capacity, the evolution of the elastic modulus G” is analyzed to identify a gradual increase in G” indicative of progressive protein aggregation corresponding to coagulation.

[0133] To assess gelling capacity, the evolution of the elastic modulus G' is analyzed to identify a net increase in G' indicating protein network formation (a jump in values ​​to a higher plateau level). The G' values ​​at 95°C and 25°C indicate the gel strength developed under hot and then cold conditions.

[0134] The results of the coagulation and gelation capacity tests are shown in Table 4.

[0135] Liquid protein extract of Chlorella vulgaris has the ability to coagulate in its pure form at a concentration of 5% (v / v) as well as when mixed with soy protein isolate in a 1:1 ratio at a concentration of 5% (v / v) as well as egg white, soy and pea protein solutions.

[0136] Liquid protein extract of Chlorella vulgaris has the ability to gel in its pure form at a concentration of 10% (v / v) and also when mixed with soy protein isolate in a 1:1 ratio at a concentration of 5% (v / v) just like egg white and soy protein solutions.

[0137] Pea protein isolate at 10% (v / v) does not exhibit gelling capacity at this concentration. A concentration higher than 10% (v / v) is required for gelling.

[0138] The elastic moduli of gels formed with the liquid protein extract of Chlorella vulgaris, at 95°C and then after cooling to 25°C, are at least equivalent to those of gels formed with egg white proteins.

[0139] The elastic moduli of the mixed gels formed with the liquid protein extract of Chlorella vulgaris mixed with soy protein isolate, at 95°C and then after cooling to 25°C, are lower than those of pure Chlorella vulgaris gels, but higher than those of pure soy protein isolate gels.

[0140] The process conditions according to example 2 of the present invention therefore make it possible to obtain a liquid protein extract having a texturizing capacity of coagulation and gelation at least equivalent to that of animal proteins (egg white proteins).

[0141] [Table 4]

[0142] Example 4: Comparison of the texturizing capacity of protein network formation within a cooked cereal product of the liquid protein extract of Chlorella vulgaris prepared according to example 2 and of egg, soy and pea proteins.

[0143] The process according to the invention is used to produce a liquid protein extract of Chlorella vulgaris according to example 2.

[0144] The ability of the protein extract to form a protein network, when mixed with wheat proteins in a cooked cereal product, is tested and compared to the ability of egg and pea proteins.

[0145] The chosen baked cereal product is a soft cake, similar to the Anglo-Saxon "layer cake." This type of soft cake is characterized by equal proportions of flour and sugar, and smaller proportions of fat, egg, and water. Baking powder, emulsifier, and salt are also used.

[0146] The application product formula, the order of incorporation of the ingredients and the parameters of the mixing phases (speed, duration) are presented in Table 5.

[0147] [Table 5]

[0148] Four cases are tested: a cake made only of wheat flour without additional protein source, a cake made of wheat flour and egg white protein, a cake made of wheat flour and liquid protein extract of Chlorella vulgaris prepared according to example 2, a cake made of wheat flour and pea protein.

[0149] The quantity of wheat flour is constant across the four cases.

[0150] The cakes for each case are made in triplicate.

[0151] A 13% (w / v) protein solution in water at pH 7 at 25°C is prepared for each additional protein source.

[0152] The samples tested in this test are presented in Table 6.

[0153] [Table 6]

[0154] The formula is made using a planetary mixer (for example a 5KSM150 model from the manufacturer Kitchen Aid, USA) equipped with the sheet tool.

[0155] Once the dough is made, it is placed in disposable aluminum molds, each containing 50g of dough. The dough is then left to rest for 30 minutes.

[0156] The cakes are then baked at 180°C for 20 minutes. Once baked, the aluminum molds are placed in individual plastic bags which are heat-sealed (using a Multivac heat sealer from Germany, for example) and then stored until further measurements are taken.

[0157] The ability of the protein extract to form a protein network when mixed with proteins from wheat flour is evaluated by measuring the size distribution of proteins present in cakes after baking by SE-HPLC.

[0158] Three cakes per batch of production are ground up and then freeze-dried, for each of the four cases studied.

[0159] A sequential extraction of proteins according to the nature of their intra and inter bonds is carried out through the sequential use of sodium dodecyl sulfate (breaking of non-covalent bonds: electrostatic, hydrogen, hydrophobic) then of dithioreythritol (breaking of disulfide bonds) combined with the use of ultrasound.

[0160] The ability of the protein extract to form a protein network in mixture with proteins from wheat flour is represented by the proportion of proteins linked by disulfide bridges, since disulfide bridges are responsible for the formation of the mixed protein network.

[0161] The greater the proportion of proteins linked by disulfide bridges within the cake, the stronger the protein network within the cake.

[0162] Protein extracts after sequential extraction are analyzed by SE-HPLC to determine the size and proportion of the protein fraction bound by non-covalent bonds, the protein fraction bound by disulfide bridges, and the inextractable protein fraction.

[0163] The references for the sequential extraction method and the SE-HPLC analysis are as follows:

[0164] Dewaest M, Villemejane C, Berland S et al (2017b) Changes in protein size distribution during wheat flour cake processing. LWT—Food Sci Technol 79:333-341.

[0165] Morel MH, Dehlon P, Autran JC et al (2000) Effects of temperature, sonication time, and power settings on size distribution and extractability of total wheat flour proteins as determined by size-exclusion high-performance liquid chromatography. Cereal Chem 77:685-691.

[0166] For each sample, the area under the curve of the elution peaks corresponding to proteins bound by non-covalent bonds, disulfide bridges, or inextractable proteins is calculated and expressed as a percentage of the total area under the curve. The total area under the curve is also compared between samples.

[0167] The results are presented in Table 7.

[0168] [Table 7]

[0169] Cakes made solely from wheat flour have a total protein content of 3% significantly lower than cakes containing wheat flour in combination with another protein source.

[0170] In cakes made solely from wheat flour, proteins linked by disulfide bonds are predominant. The proportion of inextractable (highly polymerized) proteins is significant.

[0171] Cakes made with wheat flour and egg white protein have a higher proportion of proteins linked by disulfide bonds than cakes made with wheat protein alone. The proportion of inextractable proteins is lower than in cakes made with wheat flour alone, and the same is true for the proportion of proteins linked by non-covalent bonds.

[0172] Cakes made with wheat flour and liquid protein extract of Chlorella vulgaris have a bond type distribution very similar to cakes containing egg proteins.

[0173] The proportion of proteins linked by disulfide bridges in cakes containing liquid protein extract of Chlorella vulgaris is equivalent to, or even greater than, that of cakes containing egg proteins.

[0174] Cakes made with wheat flour and pea protein have the lowest proportion of proteins linked by disulfide bonds of all the cases studied. This is mainly due to a higher proportion of proteins linked by non-covalent bonds.

[0175] The process conditions according to example 2 of the present invention therefore make it possible to obtain a liquid protein extract having a texturizing capacity of forming a mixed protein network with wheat proteins within a cooked cereal product, here a soft cake.

[0176] Example 5: Comparison of the texturizing capacity of protein network formation within a protein matrix extradited from the Tetraselmis chuii protein isolate prepared according to example 1 and egg and pea proteins.

[0177] The process according to the invention is used to produce a protein isolate of Tetraselmis chuii according to example 1.

[0178] The ability of the protein extract to form a protein network within an extradated protein matrix is ​​tested and compared to the ability of egg and pea proteins.

[0179] The chosen application matrix is ​​a protein matrix extracted by a low-water-content cooking-extrusion process. The final extracted protein matrix takes the form of low-water-content blocks (maximum 5%). w / w), with a compacted fibrous structure and some porosity. Their size is between 10 and 30 mm.

[0180] The formula of the application product is shown in Table 8.

[0181] [Table 8]

[0182] Three protein sources are tested: Tetraselmis chuii protein isolate prepared according to example 1, egg white protein, pea protein.

[0183] The protein sources are presented in Table 9.

[0184] [Table 9]

[0185] Three batches of cooking-extrusion of the formula presented in Table 8 are carried out per protein source.

[0186] The cooking-extrusion process is carried out using a twin-screw cooker-extruder of the Evolum model EV032 type (Clextral, France), Coperion ZSK (Coperion, USA) or Thermo Scientific Process 11 (Thermo Scientific, Germany).

[0187] The resulting product is fed into a twin-screw extruder at a rate of 15 kg per minute. The motor driving the extruder screws is set to a speed of 500 rpm. The temperatures of the successive extruder zones are set to 40 / 60 / 80 / 90 / 110 / 130 °C. The extruder's discharge nozzle, through which the product is ejected, is rectangular and measures 20 mm wide and 15 mm high.

[0188] The product obtained at the extruder outlet is cut into 30 mm long portions.

[0189] The extruded protein blocks obtained are then oven-dried at 40°C for 24 hours to ensure a final water content of 5% (w / v) or less. They are then packaged in heat-sealed plastic bags and stored until further measurements are carried out.

[0190] The ability of the protein isolate to form a protein network is evaluated by measuring the size distribution of proteins present in the extruded protein material by SE-HPLC following the method presented in Example 4.

[0191] 500g of extruded protein material are ground and then freeze-dried per manufacturing batch, for each of the three cases studied.

[0192] A sequential extraction of proteins according to the nature of their intra and inter bonds is carried out through the sequential use of sodium dodecyl sulfate and then dithiroreythritol combined with the use of ultrasound, following the method presented in Example 4.

[0193] The ability of the protein extract to form a protein network within the extruded matrix is ​​represented by the proportion of proteins linked by disulfide bridges.

[0194] The greater the proportion of proteins linked by disulfide bridges within the extruded protein matrix, the stronger the protein network within the cake.

[0195] Protein extracts after sequential extraction are analyzed by SE-HPLC to determine the size and proportion of the protein fraction bound by non-covalent bonds, the protein fraction bound by disulfide bridges, and the inextractable protein fraction.

[0196] The extruded protein material based on Tetraselmis chuii protein isolate has a proportion of disulfide-bound proteins at least equivalent to that of the extruded protein material based on egg white proteins.

[0197] Extruded protein material based on pea protein has the lowest proportion of proteins linked by disulfide bridges.

[0198] The process conditions according to example 1 of the present invention therefore make it possible to obtain a protein isolate having a texturizing capacity for forming a protein network within an extruded protein matrix.

Claims

Demands

1. A process for obtaining protein extract(s) with texturizing capacity from microalgae comprising at least the successive steps of: a. Preparing an aqueous suspension of microalgae with a concentration of between 1% and 20% w / w of microalgae, and a pH of between 7 and 12, at a temperature of between 4 and 25°C; c. Rupture of the rigid outer cell wall of the microalgae, preferably at a temperature of between 4 and 25°C by treating the aqueous suspension by application of ultrasound, by high-pressure homogenization or using a ball mill; d. Extraction of the aqueous protein phase from the liquid extract of microalgae with ruptured cell walls, preferably at a temperature of between 4 and 25°C by centrifugation or three-phase solvent separation; o.Extraction of a protein concentrate from the aqueous protein phase, preferably by isoelectric precipitation, precipitation by variation of ionic strength, tangential ultrafiltration, cryoprecipitation or a combination of these techniques; p. Dilution of the protein concentrate in an aqueous phase in a ratio of between 1:1 and 1:5 v / v, preferably under the same conditions of pH, ionic strength and temperature used in step o. ;.

2. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, comprising between step a. and step c. a step b. of homogenizing the aqueous suspension of microalgae obtained in step a. at a temperature between 4 and 25°C.

3. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to any one of the preceding claims, comprising after step d., a step e. of diluting the protein extract obtained in step d. in an aqueous phase in a ratio of between 1:1 and 1:10 v / v of the initial volume of the protein extract, at a pH between 7 and 12 and at a temperature between 4 and 25°C.

4. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, comprising after step d. or step e. if present, a step f of extraction of the carbohydrate fraction contained in the aqueous protein phase obtained in step d. or e. by solid / liquid separation or centrifugation, at a temperature between 4 and 25°C.

5. A process for obtaining texturizing protein extract(s) from microalgae according to the preceding claim, comprising the steps of: g. mixing the carbohydrate fraction obtained in step f. in an aqueous phase in a ratio of 1:2 to 1:10 v / v, at a pH of 7 to 12 at a temperature of 4 to 25°C; h. centrifuging and / or filtering the aqueous suspension of carbohydrate fraction obtained in step g., at a temperature of 4 to 25°C; i. mixing the residual aqueous protein extract obtained in step h. with the residual aqueous protein phase obtained in step d. or f.

6. A process for obtaining texturizing protein extract(s) from microalgae according to the preceding claim, comprising the steps of: j. mixing the aqueous protein phase obtained in step d, e, f or i with transglutaminase at a concentration of between 5 and 50 units / g of protein, at a temperature of between 25 and 55°C, for 10 to 120 minutes; k. inactivation of the transglutaminase in the mixture obtained in step j by heating at a temperature of between 60 and 80°C; l. centrifugation of the mixture obtained in step k at a temperature of between 4 and 55°C.

7. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, comprising the steps of: m. purification of the aqueous protein phase obtained in step d., e., f., i. or 1. by dialysis or adsorption on carbon particles, at a temperature between 4 and 25°C; n. centrifugation of the purified aqueous protein phase obtained in step m. at a temperature between 25 and 55°C.

8. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to any one of the preceding claims, comprising after step p., a step q. of centrifugation of the aqueous protein phase obtained in step p., preferably under the temperature conditions used in step o.

9. A process for obtaining texturizing protein extract(s) from microalgae according to the preceding claim, comprising a step r. of filtration of the aqueous protein phase obtained in step p. or q. by ultrafiltration, at a temperature between 4 and 25°C.

10. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, comprising a step s of diluting the aqueous protein phase obtained in step p., q. or r. with water at a pH between 7 and 12 and at a temperature between 4 and 25°C, so as to obtain a water content between 70 and 95% v / v.

11. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, comprising a step t. of flash-pasteurization of the aqueous protein phase obtained in step p., q., r. or s. at a temperature between 70 and 75°C for 15 to 25 seconds, so as to obtain a liquid protein extract with a water content between 65 and 95% v / v.

12. A process for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, comprising a step u. of drying the aqueous protein phase obtained in step p., q., r., s. or t. by freeze-drying, spray drying, low temperature drying or pressure drying, preferably by spray drying or freeze-drying, so as to obtain a protein extract in powder form with a water content less than or equal to 5% w / w.

13. A method for obtaining protein extract(s) with texturizing capacity from microalgae according to any one of the preceding claims, wherein said microalgae are selected from: Chlorella, Chlorella sp., Chlorella vulgaris, Chlorella protothecoides, Chlorella sorokiniana, Chlorella pyrenoidosa, Euglena sp., Euglena gracilis, Arthrospira platensis, Limnospira platensis, Spirulina maxima, Tetraselmis sp., Nannochlo-ropsis sp., Dunaliella sp., Scenedesmus sp., Aphanizomenon sp., Haematococcus sp., Tetraselmis chuii.

14. A method for obtaining protein extract(s) with texturizing capacity from microalgae according to the preceding claim, wherein said microalgae are selected from Chlorella vulgaris and Tretraselmis chuii.

15. Use of a protein extract obtained according to the process according to any one of the preceding claims as a substitute for texturizing animal proteins in food products.

Images