Method for enriching biomass with protein

Abstract

To provide a method for the protein enrichment of biomass of red unicellular algae such as Galdieria, and also to provide biomass obtained using the method.SOLUTION: A method for producing protein-rich biomass of lysed microorganisms comprises the steps of b.1) suspending biomass of lysed microorganisms, b.2) acidifying the resultant suspension to pH of 5 or less, c.) recovering the lysed biomass fraction insoluble at pH of 5 or less, and if need be, steps for repeating b.1), b.2) and c.) on the lysed biomass fraction insoluble at pH of 5 or less recovered in c.).SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a method for enriching biomass of unicellular red algae, such as Galdieria, with proteins and to the biomass obtained by said method. [Background technology]

[0002] Several plant protein sources are known to be used directly in foods for human or animal consumption or as dietary supplements to provide animals and humans with the amino acids necessary for their metabolism, and these protein sources are intended to be a source of amino acids available to animals or humans once ingested as food.

[0003] The most well-known plant protein source used in animal feed is soybeans, generally in the form of meal (the solid residue remaining after oil extraction). However, the use of soybean meal has several drawbacks related to its origin. Soybean meal is generally imported from countries where intensive soy cultivation is practiced at the expense of other plants that provide biodiversity. Furthermore, many countries encourage the cultivation of genetically modified (GM) soybean varieties, which are known to be mixed with non-GM soybeans in meal, which does not help meet the growing demand for plant-based food products that are free of genetically modified organisms (GMOs).

[0004] Other sources of plant protein, particularly Spirulina or Chlorella, are known to be used as human dietary supplements.

[0005] However, like Chlorella, Spirulina has the disadvantage of low productivity and the inability to cultivate high-yielding fermentation cultures. While these cultures can meet localized and limited demand for traditional dietary supplements, they do not achieve the goal of widespread, commercially viable industrial production of a quality dietary protein source that can replace common sources such as soy in foods for animal and human consumption.

[0006] Microalgae such as Galdieria have also been studied for their nutritional properties (Graziani et al., 2013). Protein extracts from microalgae such as Aurantiochytrium, which are known to produce oils rich in polyunsaturated fatty acids, have also been described (WO2017 / 132407, WO2017 / 019125, WO2016 / 015013). However, these extractions were performed on defatted biomass, i.e., after extraction and removal of the lipid fraction from the cultured biomass. This is also performed to recover Chlorella proteins (WO2016 / 120548, WO2016 / 00194).

[0007] For example, methods are known for producing protein-rich microbial biomass through the selection of strains particularly suited to protein overexpression and / or the implementation of cultivation methods that promote this overexpression and enrich the biomass with protein. However, biomass derived from genetically modified microorganisms (GMOs) is not suitable for certain food and feed markets, including aquaculture (particularly in Europe).

[0008] There is interest in proposing protein-rich non-GMO biomass sources as alternatives to traditional protein sources such as soy, with the goal of widespread, commercially viable industrial production. Summary of the Invention

[0009] The present invention relates to a method for producing protein-rich biomass, the method comprising: b.1. suspending the lysed microbial biomass; b.2. acidifying the resulting suspension to a pH of 5 or less; c. Recovering the dissolved biomass fraction that is insoluble at a pH of 5 or less; If necessary, repeating steps b.1, b.2, and c. on the dissolved biomass fraction insoluble at pH 5 or less recovered in step c.; Includes:

[0010] The biomass fraction recovered in step c. may then be dried.

[0011] The invention also relates to the biomass thus obtained, the use of said biomass in food products for human or animal consumption, and to food products and food compositions containing said biomass. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 shows the protein content (% of amino acids in dry matter) of Spirulina biomass and Galdieria sulfuraria strain UTEX2919 biomass before and after one cycle of treatment (HP1) or four cycles of treatment (HP4) according to the present invention. [Figure 2] FIG. 1 shows the amino acid composition (g / 100 g dry matter) of Spirulina biomass and Galdieria sulfuraria strain UTEX2919 biomass before and after one cycle of treatment (HP1) or four cycles of treatment (HP4) according to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0013] The present invention provides a method for producing lysed microbial protein-rich biomass, the method comprising: b.1. suspending the lysed microbial biomass; b.2. acidifying the resulting suspension to a pH of 5 or less; c. Recovering the dissolved biomass fraction that is insoluble at a pH of 5 or less; If necessary, repeating steps b.1, b.2, and c. on the dissolved biomass fraction insoluble at pH 5 or less recovered in step c.; The present invention relates to a method, comprising:

[0014] According to the invention, the microorganisms are essentially microorganisms that accumulate water-soluble glycogen, such as yeasts or protists, which accumulate water-soluble glycogen during growth, in particular under industrial conditions of fermentation culture with biomass accumulation.

[0015] Advantageously, the method of the present invention is particularly suitable for biomass of phycobiliprotein-producing microorganisms, and advantageously adds value to the biomass remaining after extraction of the phycobiliproteins, i.e., a by-product of the production of these molecules of interest.

[0016] These microorganisms and methods for culturing them are well known to those skilled in the art and are described in particular in WO2017 / 050917, WO2017 / 050918 and WO2017 / 093345.

[0017] In particular, unicellular red algae (URA) are mentioned. According to the present invention, "URA" refers to the Rhodophytes, in particular to the subphylum Cyanidiophytina, preferably to the class Cyanidiophyceae, in particular to the order Cyanidiales, more particularly to the family Cyanidiaceae or Galdieriaceae, and even more particularly to the genera Cyanidioschyzon, Cyanidium or Galdieria.

[0018] Preferably, the URA is selected from the species Cyanidioschyzon merolae 10D, Cyanidioschyzon merolae DBV201, Cyanidium caldarium, Cyanidium daedalum, Cyanidium maximum, Cyanidium partitum, Cyanidium rumpens, Galdieria daedala, Galdieria maxima, Galdieria partita or Galdieria sulphuraria, more preferentially Galdieria sulphuraria.

[0019] Preferably, the method of the invention is carried out with a lysed biomass in which at least 70% of the cells have been lysed, more preferentially at least 80% of the cells have been lysed, even more preferentially 90% of the cells have been lysed, even more preferentially 95% of the cells have been lysed.

[0020] According to one embodiment of the invention, the method of the invention comprises a preliminary step a. of lysing the pre-cultured microbial biomass.

[0021] The lysis step a. may be carried out by any cell lysis means known to those skilled in the art, in particular by grinding or enzymatic lysis. Preferably, the cell lysis is carried out by grinding, which is particularly preferred for the lysis of URA, in particular Galdieria sulphuraria species.

[0022] Thus, step a. can be carried out while the cells are suspended in water, in the fermentation must or in a reconstituted suspension, or on so-called "dry" biomass, i.e. on fresh biomass separated from the fermentation medium, or on biomass after drying and storage, e.g. after freeze-drying.

[0023] Advantageously, lysis by comminution is carried out on "dry" cell biomass, in particular on fresh cell biomass separated from the fermentation medium after cultivation, by any separation method known to those skilled in the art, in particular by filtration or centrifugation.

[0024] According to another embodiment of the invention, lysis by grinding is performed on a cell biomass that has previously been freeze-dried or dried for storage.

[0025] The suspension step b.1. makes it possible to obtain a suspension of dissolved biomass.

[0026] The suspension obtained in step b1 generally contains 1% to 40% or more dry matter, preferably 20% to 30% dry matter.

[0027] Step b.2. is carried out by adding an acid to obtain a suspension in which the pH of the aqueous phase of the suspension is less than or equal to 5. Preferably, a pH of less than or equal to 4 is sought.

[0028] The pH is adjusted by adding an acid, either inorganic or organic, strong or weak, in the form of a solid or solution, and the amount of acid added is determined by the pH of the suspension to be treated and the pH value that the skilled person wants to achieve. Among the inorganic acids known to those skilled in the art, hydrochloric acid, sulfuric acid, and phosphoric acid are particularly mentioned. Among the organic acids known to those skilled in the art, acetic acid, citric acid, tartaric acid, and lactic acid are particularly mentioned, with citric acid being preferred. Also included are acidic polyphenols such as rosmarinic acid, tannic acid, digallic acid, quercitic acid, gallotannic acid, acidic tannins such as quercetin, ellagitannin, castalagin, castalin, casuariticin, grandinin, punicaligin, punicalin, robulin A, tellimagrandin II, terflavin B, vescaligin, pendunculagin, casuariin, castrin, vescalin, preferably tannic acid. Preferably, the acid used is a food-approved acid, in particular phosphoric acid, sulfuric acid, citric acid or tannic acid.

[0029] The acidic suspension obtained in step b.2. preferably has a dry matter content of greater than or equal to 1%, advantageously greater than 2%, preferentially between 3 and 30%, more preferentially between 4 and 15%, even more preferentially between 5 and 12%, even more preferentially between 6 and 10%.

[0030] According to a particular embodiment of the invention, steps b.1. and b.2. are consecutive, and step b.2. is carried out, for example, immediately after step b.1., without any other biomass treatment steps, such as a degreasing step. Following the suspension or dilution of the suspension, an acid is added in solid or liquid form.

[0031] According to another particular embodiment of the present invention, steps b.1. and b.2. are carried out simultaneously by adding an appropriate amount of acidic solution to obtain both the desired pH below 5 and the desired dry matter content.

[0032] Depending on the lysed biomass recovered from step a., in particular whether the lysis was carried out in an aqueous suspension of cells or on a so-called "dry" biomass, a person skilled in the art will know how to add the amounts of water and acid required to obtain a suspension with the desired dry matter content and the desired pH, regardless of whether these additions are carried out in two steps or in one step.

[0033] Step c. is carried out by any separation means known to those skilled in the art, in particular by filtration or centrifugation, to separate the solid residue of the lysed cells from the aqueous solution derived from the acidic suspension containing the lysed cells. Filtration is particularly tangential filtration, for example, tangential filtration on a ceramic membrane or an organic membrane such as hollow polyethersulfone fiber. Frontal filtration (plate filter or press filter) may also be used.

[0034] If necessary, the suspension / acidification / recovery cycle (step b.1. / step b.2. / step c.) may be repeated several times on the lysed biomass fraction insoluble at pH 5 or less recovered in step c. in order to deplete components soluble at acidic pH. Advantageously, the suspension / acidification / recovery cycle is repeated once, twice or three times on the lysed biomass recovered in step c.

[0035] The protein-rich biomass fraction recovered in step c. may then be dried by any known drying method.

[0036] The mother liquor of the suspension is typically recovered for processing at acidic pH to extract soluble components, which may be valuable in their own right. This is particularly true for phycocyanin when the microorganism is a phycobiliprotein-producing microorganism as described above. Such a process is described, inter alia, in patent application PCT / EP2018 / 058294, filed March 30, 2018.

[0037] The resulting biomass generally has at least 60% protein (assessed by total nitrogen determination, N x 6.25) on dry matter, preferably more than 65%, more preferentially more than 68%, more than 70% or more than 80% protein.

[0038] The biomass also preferably has a total sugar content of less than 20%, more preferentially less than 10%, less than 5% based on dry matter.

[0039] The biomass obtained also preferably has a glycogen content of less than 10%, more preferentially less than 5%, or less than 1%, and even more preferentially less than 0.1%, based on dry matter.

[0040] Depending on the microorganism used, the dissolved biomass, which is insoluble at a pH of 5 or less, may have a fat content, in particular a lipid content, which may be at least 5%, or at least 10%, or at least 15%, or at least 16%, or even up to 20% of the dry matter. The fat composition also varies depending on the microorganism. For URA, the dissolved biomass according to the invention generally comprises polyunsaturated fatty acids of the omega-3, 6 and 9 series, in particular oleic acid, palmitic acid and α-linolenic acid, which represent at least 30%, preferentially 40%, more preferentially 50% of the fat.

[0041] A person skilled in the art will know how to adapt the conditions for carrying out the method according to the invention on the recovered lysed biomass, in particular the pH of the suspension and the number of repetitions of the suspension / acidification / recovery cycle, in order to obtain a biomass containing the desired amount of proteins, sugars, glycogen and lipids, in particular the biomass described above.

[0042] The weight percent of protein may be expressed relative to the protein itself or to the amino acids contained in said protein. According to the present invention, protein means any protein, peptide or amino acid that is insoluble at an acidic pH of 5 or less.

[0043] The present invention also relates to the biomass defined above, which is obtainable by the method according to the invention.

[0044] This biomass is advantageously - at least 60% protein, preferably more than 65%, more preferentially more than 68%, more than 70% or more than 80% protein, less than 20% total sugars, preferentially less than 10%, more preferentially less than 5% total sugars; less than 10% glycogen, preferably less than 5%, preferentially less than 1% and more preferentially less than 0.1% glycogen, and - Up to 20% fat and The above percentages (%) are expressed relative to the dry matter of the biomass.

[0045] The present invention also relates to the use of said biomass in the fields of cosmetics, medicines and food for human or animal consumption.

[0046] The present invention relates in particular to the use of a protein-rich biomass according to the invention as described above or below for improving the performance of animals, which may be assessed in particular by measuring feed intake, weight gain or feed conversion ratio.

[0047] In animal feed, a distinction can be made between the feeding of livestock, especially livestock in industrial agriculture, and the feeding of farm animals or pets or so-called "leisure" animals, such as fish in aquariums or birds in aviaries or cages or exotic pets.

[0048] "Livestock" refers in particular to herbivorous animals (especially cattle raised for meat, milk, cheese and leather; sheep raised for meat, wool and cheese; goats), pigs, rabbits, poultry (chickens, hens, turkeys, ducks, geese, etc.), equines (ponies, horses, foals), aquatic animals (e.g. fish, shrimp, shellfish (especially oysters and mussels)) intended to support human activity (transport, recreation) or to provide food for humans. However, a distinction can be made between the feeding of fish up to the juvenile stage and the feeding of farmed fish, including the feeds and feed compositions intended therefor.

[0049] These domestic animals are to be distinguished from domestic animals, pets or leisure pets, including mammals, ruminants or non-ruminants, birds or fish. In particular, domestic animals include dogs and cats.

[0050] The present invention also relates to a food product or food composition for humans or animals, comprising the biomass according to the invention described above. By "food product" is meant any composition that can be used for human or animal consumption.

[0051] According to the present invention, the food product may comprise only dried or wet, treated or untreated biomass, or may be a mixture of dried or wet, treated or untreated biomass with any other additive, vehicle or carrier used in the field of food for human or animal consumption, such as food preservatives, colorants, flavor enhancers, pH adjusters, or pharmaceutical additives such as growth hormones, antibiotics, etc.

[0052] The present invention relates in particular to animal feed, more particularly to livestock feed, which is usually in the form of powder, granules or soup, to which the biomass according to the invention has been incorporated. "Animal feed" refers to anything that can be used to feed animals.

[0053] In intensive livestock production, the feed may contain a nutrient base and nutritional additives in addition to the algal biomass. Thus, the majority of the animal feed formulation consists of a "nutrient base" and algal biomass. In one example, this nutrient base is composed of a mixture of grains, proteins, and fats of animal and / or plant origin.

[0054] Nutritional bases for animals are adapted to feed these animals and are well known to those skilled in the art. In the present invention, these nutritional bases include, for example, corn, wheat, peas, and soybeans. These nutritional bases are adapted to the needs of different target animal species. These nutritional bases may already contain nutritional additives such as vitamins, mineral salts, and amino acids.

[0055] Additives used in animal feed may be added to improve certain properties of the feed, for example to enhance flavor, make the feed material more digestible, or to protect the animals, and are often used in large-scale intensive livestock production.

[0056] The additives used in animal feed are divided into the following subcategories in detail (source: EFSA): - technical additives: e.g. preservatives, antioxidants, emulsifiers, stabilizers, pH adjusters and silage additives; - organoleptic additives: e.g. flavourings, colourings; - Nutritional additives: for example, vitamins, amino acids and trace elements; - Zootechnical additives: e.g. digestive aids, intestinal flora stabilizers; - Anticoccidial and antihistomonasal (antiparasitic) agents.

[0057] In one embodiment, the present invention relates to livestock feed comprising 1 to 60%, preferably 1 to 20%, most preferentially 3 to 8% of dry biomass obtained by the method according to the invention.

[0058] In another embodiment, the present invention relates to livestock feed containing 1 to 40%, preferably 5 to 10%, of undried biomass obtained by the method of the present invention.

[0059] According to a particular embodiment of the invention, the feed is intended for livestock, in particular cattle, sheep, pigs, rabbits, poultry and equines.

[0060] According to another particular form of the invention, the food product is intended for aquatic animals, in particular fish, at least up to the juvenile stage, including farmed and ornamental fish.

[0061] According to another particular embodiment of the invention, the food product is intended for domestic animals, pets and / or leisure animals and exotic pets.

[0062] Finally, according to another embodiment of the invention, the food composition is intended for humans.

[0063] The present invention also relates to a cosmetic, nutraceutical or pharmaceutical composition for humans or animals comprising the biomass according to the invention described above.

[0064] According to the present invention, the cosmetic, nutraceutical or pharmaceutical composition may comprise only dried or wet, converted or unconverted biomass, or may be a mixture of dried or wet, converted or unconverted biomass with any other additive, vehicle or carrier used in the cosmetic or pharmaceutical field, such as preservatives, colorants, pH adjusters, etc.

[0065] The present invention also relates to the use of said biomass in the prevention and treatment and therapy of malnutrition. [Example]

[0066] material and method Dissolved Biomass The biomass treated by the method according to the present invention was biomass of Spirulina (Arthrospira platensis) and biomass of Galdieria sulphuraria strain UTEX2919 (Algal Culture Collection, University of Texas, USA, 205 West 24th Street, Biological Research Building 218, The University of Texas at Austin (A6700), Austin, TX 78712, USA), which were cultivated according to conventional methods for culturing these strains. The lysed biomass has a lysed cell content of 95%. The dry matter content is 10%.

[0067] Operating Procedure The enrichment process involves repeatedly rinsing the ground biomass pellet under acidic pH conditions. To accomplish this, the procedure involves the use of a centrifuge, as described here with a Sorvall RC5B Plus and a Sorvall SLA3000 fixed-angle rotor. The ground biomass is centrifuged at 12,000 x g for 15 minutes to maximize pelleting of the insoluble fraction. The supernatant, known as the "soluble fraction," is removed and replaced with water. After resuspending the pellet, the centrifugation procedure is repeated, monitoring the pH and adjusting it to a value below pH 4.8 as necessary. Repeating this process three to four times maximizes protein and lipid enrichment of the insoluble fraction.

[0068] analysis Total protein content is evaluated by the Kjeldahl method known to those skilled in the art.The amino acid profile by acid hydrolysis is carried out according to the method of ISO 13903:2005;EU152 / 2009.The analysis of lipid content and composition is carried out by GC / FID and internal calibration known to those skilled in the art.The sugar profile is determined by ion chromatography-pulse amperometric detection.

[0069] result The composition of the heavy liquid phase (HP, or insoluble fraction) after the first extraction (HP1) and the heavy liquid phase after the fourth extraction (HP4) was monitored and compared to the composition of the input lysate for Galdieria sulphuraria strain UTEX2919 biomass (Table 1) and Spirulina biomass (Table 2). Percentages (%) are expressed relative to dry matter (DM).

[0070] [Table 1]

[0071] The first suspension / acidification / recovery cycle resulted in a clear increase in the protein content of the lysed biomass (>6%), which was even more pronounced (>16%) after the fourth treatment cycle.

[0072] [Table 2]

[0073] Contrary to URA such as Galdieria, the process of the present invention for Spirulina resulted in a decrease in the protein content of the insoluble dissolved biomass at pH 5 and below (Table 2). For extraction at pH 4, which is lower than the pH of allophycocyanin (APC) and phycocyanin (C-PC), a decrease in the amount of phycocyanin and allophycocyanin in the heavy liquid phase was observed. Since C-PC and APC are insoluble in water at pH 4, the loss observed with HP4 for this biomass is apparently due to the partial decomposition of these molecules at this pH.

[0074] The protein content of the different biomasses is shown in Figure 1. A closer look at the amino acid composition of the different products, shown in Figure 2, reveals an overall increase in all amino acids in the heavy liquid phase, with amounts of most amino acids, except alanine, equal to or greater than those of spirulina.

[0075] Considering the FAO recommendations on essential amino acids for human nutrition, and comparing the compositions of the different heavy liquid phases, it is observed that their amounts are all above the recommended values.

[0076] After several extractions, phycocyanin (C-PC and APC) still remained in the heavy liquid phase, accounting for approximately 3% of the dry mass. Phycocyanin is known to be a molecule with strong antioxidant properties. The fatty acid composition revealed that omega-3, omega-6, and omega-9 unsaturated fatty acids, equivalent to over 50% of the fat mass, were present in the form of polar lipids, accounting for approximately 5-20% of the dry mass of the heavy liquid phase (Table 3).

[0077] [Table 3]

[0078] Sugars, like fiber, are below the detection limit of the method of the present invention (Table 4).

[0079] [Table 4]

[0080] The method according to the invention was repeated on Galdieria sulphuraria strain UTEX2919 biomass cultivated according to the method described in patent application WO2017 / 050917.

[0081] The starting biomass contains more than 70% protein by DM. After three rounds of treatment according to the invention, a biomass containing more than 80% protein is obtained, an increase of more than 14%.

[0082] Successive extractions were performed on three biomasses with different glycogen contents. It is noteworthy that almost all of the soluble glycogen in the biomass was found in solution after the first extraction.

[0083] [Table 5]

[0084] The protein-rich heavy liquid phase thus obtained is then enzymatically treated with a protease to obtain a protein hydrolysate, increasing the solubility and digestibility of the product.

[0085] The recommended amount of protein varies for each person at different stages of life and with regard to daily energy expenditure (ANSES, 2007). For healthy adults, the recommended dietary allowance (RDA) for protein is 0.83 g / kg / day. However, these requirements vary and are increased for pregnant women, the elderly and certain athletes.

[0086] The protein-rich fraction produced by the method of the present invention may be ingested directly in the form of a cold-pressed tablet, a powder in capsule form, or as a powder to be mixed with other ingredients. The fraction may also be incorporated into other food preparations, such as those used in the manufacture of pancakes, cookies, other pastries, or cereal bars, or may be incorporated into any preparation as a substitute for conventional flour, in order to enrich the food product thus prepared with protein.

[0087] The protein fractions can also be envisaged for use in preparations based on milk (animal or vegetable), raw milk or fermented milk, such as cream desserts, yogurt, cheese, etc. By adding the protein fractions, it is possible to obtain protein-rich milk or cheese preparations while reducing the amount of fat. The protein fractions can also be envisaged for making preparations based on raw cheese, pressed or unpressed cheese, or processed cheese. The algae can be incorporated into the mass of the preparation, or simply on the outside of the rind if the rind of the preparation is edible.

[0088] This protein fraction may also be incorporated into preparations for use as meat substitutes such as veggie burgers, or into surimi-style protein sticks, in which case the proteins may be textured or used as is.

[0089] Another example use is in powder form to be added to beverages such as smoothies or shakes.

[0090] Composition example: Algae protein enriched flour: Composition of 1 kg of flour: 800g flour (10-12% protein) 200g protein-enriched seaweed extract (65-80% protein) That is, 16-19% protein per 100g of enriched flour compared to 10-12% protein in regular Type 55 flour.

[0091] Cookies made with algae protein-enriched flour Ingredients per 100g of cookies (7g-9g protein): 45g seaweed protein enriched flour 23g butter 21.7g sugar 5g cocoa powder 4g of eggs 1g of salt 0.3g baking powder

[0092] Bread made with algae protein-enriched flour Ingredients per 100g of bread (10-25g protein): 63.2g seaweed protein enriched flour 34g water 1.8g salt 1g baker's yeast

[0093] Seaweed veggie burger Composition of 100g of burger Water, chickpea flour (26%), seaweed powder (25%), onion, carrot, wheat gluten (3%), lemon juice, yeast extract, potato starch, vegetable oil (sunflower), turmeric, garlic, pH adjuster (potassium chloride), antioxidant, potato fibre, thickener (carrageenan), starch (corn, wheat), emulsifier (guar gum). This equates to 22-33g of protein per burger.

[0094] Algae protein-enriched dairy products Gouda-type cheese-based preparations The protein-enriched seaweed fraction is added to the coagulated milk protein, and the resulting paste is then pressed. The proportion of protein-enriched seaweed fraction can vary from 1% to 50%. In this case, the limiting factor relates to the appearance of the final product, especially its texture and cuttability.

[0095] [Table 6]

[0096] Algae protein-enriched beverage powder Smoothie composition: 93.8g of powder derived from an algae protein enriched fraction 5g natural fruit flavors 0.25g xanthan gum 0.2g guar gum 0.25g of natural sweeteners or flavorings with sweetening power Mix 25g of powder with 200ml of cold water or fruit juice. This equates to 15-18.5g of protein per serving.

[0097] Shake composition: 84.35g of powder derived from algae protein enriched fraction 10g milk powder 5g cocoa 0.2g xanthan gum 0.2g guar gum 0.25g sweetener Mix 25g of powder in 200ml of cold or hot water, which equates to 15-20g of protein per serving.

[0098] References WO2016 / 00194 WO2016 / 120548 WO2016 / 015013 WO2017 / 019125 WO2017 / 050917 WO2017 / 050918 WO2017 / 093345 WO2017 / 050917 WO2017 / 132407 US2017-020834 Graziani & al., Food & Function,vol.4,n°1,2013,144-152.

Claims

1. 1. A method for producing lysed microbial protein-rich biomass, the method comprising: b.

1. suspending the lysed microbial biomass; b.

2. acidifying the resulting suspension to a pH of 5 or less; c. Recovering the dissolved biomass fraction that is insoluble at pH 5 or less; If necessary, repeating steps b.1, b.2 and c on the dissolved biomass fraction insoluble at pH 5 or less recovered in step c. A method comprising:

2. 2. The process according to claim 1, characterized in that the pH of the acidic suspension obtained in step b.

2. is below 4.

5.

3. 3. The process according to claim 1 or 2, characterized in that the dry matter content of the acidic suspension obtained in step b.

2. is between 1% and 30%.

4. 4. The method according to any one of claims 1 to 3, characterized in that steps b.

1. and b.

2. are successive or simultaneous.

5. 5. The method according to any one of claims 1 to 4, characterized in that steps b.

1. and b.

2. are consecutive and step b.

2. is carried out immediately after step b.

1. without further biomass treatment steps, in particular without a defatting step.

6. 6. The method according to any one of claims 1 to 5, characterized in that it comprises a preliminary step a. of lysing a pre-cultured microbial biomass.

7. 7. The method according to any one of claims 1 to 6, characterized in that the biomass recovered in step c. is dried.

8. 8. The method according to any one of claims 1 to 7, characterized in that the microorganism is selected from unicellular red algae (URA) of the families Cyanidiaceae or Galdieriaceae.

9. 9. A protein-rich lysed microbial biomass obtainable by the method according to any one of claims 1 to 8.

10. 10. Biomass according to claim 9, characterized in that it contains at least 60% protein by dry matter and has a total sugar content of at least less than 20% by dry matter and / or a glycogen content of less than 10% by dry matter and / or a fat content of at least 5% by dry matter.

11. 11. A food or food composition for human or animal consumption, characterized in that it comprises a lysed microbial biomass according to claim 9 or 10.

12. 11. A lysed microbial biomass according to claim 9 or 10 for therapeutic use.

13. 11. The lysed microbial biomass according to claim 9 or 10 for use in the prevention and treatment of malnutrition.

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