# US 11,560,542 B2 · Process for producing phycocyanin-rich biomass via URA culturing > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,560,542 B2 - Document ID: US11560542B2 - Status: Granted - Kind: B2 - Application number: 17/059,979 - Filed: 2019-05-27 - Published: 2021-07-29 - Granted: 2023-01-24 - Earliest claimed: 2018-05-31 - Assignees: Fermentalg - Applicants: Fermentalg - Inventors: Olivier Cagnac; Axel Athane; Marion Champeaud - Web page: https://patentgrep.com/patent/US11560542B2 ## CPC - C12N 1/12: Unicellular algae; Culture media therefor (as new plants A01H13/00) - C12P 17/165: {Heterorings having nitrogen atoms as the only ring heteroatoms} - A01G 33/00: Cultivation of seaweed {or algae} - A23L 5/46: using dyes or pigments of microbial or algal origin - C07K 14/405: from algae - C12N 1/32: Processes using, or culture media containing, lower alkanols, i.e. C1 to C6 - C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione - C12P 23/00: Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes (containing heterorings C12P17/00) - A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs - C12N 2500/34: Sugars - Y02A 40/80: in fisheries management - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)} - A01G: HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING (picking of fruits, vegetables, hops or the like A01D46/00; propagating unicellular algae C12N1/12) - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) - A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION - Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE ## Abstract The invention relates to the cultivation of unicellular red algae (URA) for producing biomass for the production of products of interest, such as dry biomass or compounds or mixtures of compounds of interest extracted from the biomass produced, particularly food pigments or colouring agents. The invention more particularly relates to the industrial production of said biomass, which must satisfy an economic equilibrium of profitability, with both an increase in productivity (quantity of biomass and of compounds of interest in the biomass) and an economically acceptable production cost. ## Claims ### Claim 1 A process for production of a phycocyanin-rich biomass, the process comprising: (i) cultivating unicellular red algae (URA) in mixotrophic or heterotrophic mode on a culture medium comprising a carbon source, the carbon source comprising glucose, wherein the cultivating comprises a first step of growth to produce biomass and a second step of accumulation of phycocyanin, and wherein glycerol is added in the culture medium, and (ii) recovering the biomass, wherein the weight ratio of glycerol to carbon source comprising glucose (Gly/Glu) after addition of glycerol is of at least 1/15. ### Claim 2 (depends on 1) The process according to claim 1, wherein the URA is of the genera Cyanidioschyzon, Cyanidium or Galdieria. ### Claim 3 (depends on 1) The process according to claim 1, wherein the carbon source in the culture medium is selected from the group consisting of: glucose, lactose, fructose or glucose-containing polysaccharides, and mixtures thereof. ### Claim 4 (depends on 1) The process according to claim 1, wherein the carbon source is lactose from milk permeate. ### Claim 5 (depends on 1) The process according to claim 1, wherein the culture medium comprises between 0.05 g/L and 200 g/L of the carbon source comprising glucose. ### Claim 6 (depends on 1) The process according to claim 1, wherein the cultivating is in mixotrophic mode, and further comprises continuous or discontinuous illumination. ### Claim 7 (depends on 1) The process according to claim 1, wherein the addition of glycerol to the culture medium is carried out at the beginning of the culture. ### Claim 8 (depends on 1) The process according to claim 1, wherein the weight ratio of glycerol to carbon source comprising glucose (Gly/Glu) is at least 1/14. ### Claim 9 (depends on 1) The process according to claim 1, wherein the weight ratio of glycerol to carbon source comprising glucose (Gly/Glu) is at least 1/10. ### Claim 10 (depends on 1) The process according to claim 1, wherein the weight ratio of glycerol to carbon source comprising glucose (Gly/Glu) is at least 1/5. ### Claim 11 (depends on 1) The process according to claim 1, wherein the weight ratio of glycerol to carbon source comprising glucose (Gly/Glu) is from 1/5 to 1/3. ### Claim 12 A process for preparing phycocyanins, the process comprising: (i) cultivating unicellular red algae (URA) in mixotrophic or heterotrophic mode on a culture medium comprising a carbon source, the carbon source comprising glucose, wherein the cultivating comprises a first step of growth to produce biomass and a second step of accumulation of phycocyanin, and wherein glycerol is added in the culture medium, (ii) recovering the biomass, wherein the weight ratio of glycerol to carbon source comprising glucose (Gly/Glu) after addition of glycerol is of at least 1/15, and (iii) extracting the phycocyanins from the biomass. ### Claim 13 (depends on 1) The process according to claim 1, wherein the URA is selected among 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. ### Claim 14 (depends on 1) The process according to claim 1, wherein the addition of glycerol to the culture medium is carried out once the biomass has reached a density greater than 20 g/L dry matter. ### Claim 15 (depends on 1) The process according to claim 1, wherein the biomass recovered has a phycocyanin content of between 1 and 250 mg/g dry matter. ### Claim 16 (depends on 1) The process according to claim 1, wherein the biomass recovered has a phycocyanin content of between 0.5 and 100 mg/g dry matter. ### Claim 17 (depends on 1) The process according to claim 1, wherein the biomass recovered has a carotenoid content between 0.1 and 10 mg/g dry matter.