# US 2020/0399526 A1 · Co-cultivation of a myxobacterium and acinetobacter for enhanced production of emulsan > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399526 A1 - Document ID: US20200399526A1 - Status: Published application - Kind: A1 - Application number: 16/905,170 - Filed: 2020-06-18 - Published: 2020-12-24 - Earliest claimed: 2019-06-20 - Assignees: Locus IP Company, LLC - Applicants: Locus IP Company, LLC - Inventors: SEAN FARMER; KEN ALIBEK; YAJIE CHEN - Web page: https://patentgrep.com/patent/US20200399526A1 ## CPC - C09K 8/582: characterised by the use of bacteria - C12N 1/20: Bacteria; Culture media therefor - C12P 5/02: acyclic {(C12P5/007 takes precedence)} - C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil - C12P 19/04: Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds - C12P 19/26: Preparation of nitrogen-containing carbohydrates - C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione - C12P 39/00: Processes involving microorganisms of different genera in the same process, simultaneously - C09K: MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE - 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)} ## Abstract Methods are provided for enhanced production of one or more microbial biopolymers, the methods comprising co-cultivating Stigmatella aurantiaca and Acinetobacter venetianus. In certain embodiments, the one or more biopolymers are emulsan. In certain embodiments, other microbial growth by-products are produced, such as biosurfactants. Microbe-based products produced according to the subject methods are also provided, as well as their uses in, for example, oil and gas recovery, agriculture, and health care. ## Claims ### Claim 1 A method for enhanced production of one or more microbial growth by-products, the method comprising co-cultivating a first microorganism and a second microorganism in a fermentation reactor, wherein the first microorganism is a myxobacterium and the second microorganism is a strain of Acinetobacter, and wherein a greater concentration of the one or more microbial growth by-products is achieved than would be achieved if the first and second microorganisms were cultivated individually. ### Claim 2 (depends on 1) The method of claim 1, wherein the myxobacterium is a Stigmatella spp. bacterium ### Claim 3 (depends on 2) The method of claim 2, wherein the Stigmatella spp. bacterium is S. aurantiaca. ### Claim 4 (depends on 1) The method of claim 1, wherein the Acinetobacter is a strain of A. venetianus. ### Claim 5 (depends on 4) The method of claim 4, wherein the strain is RAG-1. ### Claim 6 (depends on 1) The method of claim 1, wherein the myxobacterium is S. aurantiaca and the Acinetobacter is A. venetianus. ### Claim 7 (depends on 1) The method of claim 1, wherein the Acinetobacter produces one or more growth by-products. ### Claim 8 (depends on 1) The method of claim 1, wherein the one or more growth by-products are biopolymers. ### Claim 9 (depends on 8) The method of claim 8, wherein at least one of the one or more biopolymers is emulsan. ### Claim 10 (depends on 1) The method of claim 1, wherein the one or more growth by-products are biosurfactants. ### Claim 11 (depends on 10) The method of claim 10, wherein the biosurfactants are glycolipids. ### Claim 12 (depends on 10) The method of claim 10, wherein the biosurfactants are lipopeptides. ### Claim 13 (depends on 1) The method of claim 1, wherein co-cultivating the first and the second microorganisms comprises: inoculating the fermentation reactor with the first microorganism and inoculating the fermentation reactor with the second microorganism, wherein the fermentation reactor comprises a liquid nutrient medium; incubating the first and second microorganisms in the reactor under conditions favorable for growth and production of the one or more microbial growth by-products; extracting the one or more growth by-products from the reactor; and, optionally, purifying the one or more growth by-products. ### Claim 14 (depends on 13) The method of claim 13, wherein the liquid nutrient medium comprises sucrose, casein hydrolysate, magnesium sulfate, dipotassium phosphate, monopotassium phosphate, calcium chloride, ammonium sulfate, trace metals, and water. ### Claim 15 (depends on 13) The method of claim 13, further comprising suspending a particulate anchoring carrier in the liquid nutrient medium. ### Claim 16 (depends on 15) The method of claim 15, wherein the first and/or second microorganism attaches to the particulate anchoring carrier and accumulates thereon in the form of a biofilm to form a plurality of bacterial-carrier masses. ### Claim 17 (depends on 13) The method of claim 13, further comprising adding an aqueous base solution comprising 15 to 25% NaOH and/or an aqueous acid solution comprising 15 to 25% citric acid to the reactor. ### Claim 18 (depends on 13) The method of claim 13, further comprising feeding 7% of clear liquefied paraffin into the reactor after 24 hours of incubation. ### Claim 19 (depends on 13) The method of claim 13, further comprising feeding 6% oil-based defoamer into the reactor every 24 hours. ### Claim 20 (depends on 1) The method of claim 1, wherein the first microorganism stimulates enhanced production of the one or more growth by-products by the second microorganism. ### Claim 21 A composition comprising two or more microorganisms and one or more microbial growth by-products, said microorganisms comprising Stigmatella aurantiaca and Acinetobacter venetianus, and said one or more microbial growth by-products comprising at least one biopolymer. ### Claim 22 (depends on 21) The composition of claim 21, wherein the biopolymer is emulsan. ### Claim 23 (depends on 21) The composition of claim 21, wherein the Acinetobacter is strain RAG-1.