# US 11,297,868 B2 · Method of generating bacterial compositions > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 11,297,868 B2 - Document ID: US11297868B2 - Status: Granted - Kind: B2 - Application number: 16/304,697 - Filed: 2017-05-29 - Published: 2019-07-18 - Granted: 2022-04-12 - Earliest claimed: 2016-05-29 - Assignees: The State of Israel, Ministry of Agriculture & Rural Development, Agricultural Research Organization (ARO) (Volcani Center); Yissum Research Development Company of the Hebrew University of Jerusalem - Applicants: The State of Israel, Ministry of Agriculture & Rural Development, Agricultural Research Organization (ARO) (Volcani Center); Yissum Research Development Company of the Hebrew University of Jerusalem Ltd. - Inventors: Moshe Shemesh; Ram Reifen - Web page: https://patentgrep.com/patent/US11297868B2 ## CPC - A23L 33/135: Bacteria or derivatives thereof, e.g. probiotics - A61K 35/742: Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes - A61K 35/747: Lactobacilli, e.g. L. acidophilus or L. brevis - A61P 1/14: Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents - A61P 3/06: Antihyperlipidemics - A61P 31/04: Antibacterial agents - A61P 37/02: Immunomodulators - A61P 39/02: Antidotes - C12N 1/20: Bacteria; Culture media therefor - C12N 1/205: Bacterial isolates - C12P 39/00: Processes involving microorganisms of different genera in the same process, simultaneously - C12R 2001/125: Bacillus subtilis {; Hay bacillus; Grass bacillus} - C12R 2001/25: Lactobacillus plantarum - A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - 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)} - C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS ## Abstract A method of preparing a bacterial composition is disclosed. The method comprises: (a) in vitro co-culturing beneficial bacteria with biofilm-producing bacteria in a growth substrate under conditions that generate a biofilm which comprises the beneficial bacteria and the non-pathogenic bacteria; and (b) isolating the biofilm from the growth substrate. ## Claims ### Claim 1 A method of preparing a bacterial composition comprising: (a) in vitro co-culturing beneficial bacteria with biofilm-producing bacteria in a growth substrate under conditions that generate a biofilm which comprises said beneficial bacteria and said biofilm-producing bacteria; and (b) isolating said biofilm from said growth substrate, thereby preparing the bacterial composition. ### Claim 2 (depends on 1) The method of claim 1, wherein said biofilm-producing bacteria are non-pathogenic bacteria. ### Claim 3 (depends on 1) The method of claim 1, wherein said biofilm-producing bacteria are of the Bacillus genus. ### Claim 4 (depends on 3) The method of claim 3, wherein said biofilm-producing bacteria are of the B. subtilis species. ### Claim 5 (depends on 1) The method of claim 1, wherein said beneficial bacteria are probiotic bacteria. ### Claim 6 (depends on 5) The method of claim 5, wherein said probiotic bacteria are of the Lactobacillales order. ### Claim 7 (depends on 5) The method of claim 5, wherein said biofilm-producing bacteria are of the B. subtilis species. ### Claim 8 (depends on 5) The method of claim 5, wherein said probiotic bacteria are of the L. plantarum species. ### Claim 9 (depends on 1) The method of claim 1, wherein said beneficial bacteria are genetically modified to express a therapeutic polypeptide. ### Claim 10 (depends on 1) The method of claim 1, wherein said beneficial bacteria are used in bioremediation. ### Claim 11 (depends on 1) The method of claim 1, wherein said biofilm-producing bacteria express genes of the KinD-Spo0A pathway. ### Claim 12 (depends on 11) The method of claim 11, wherein said growth substrate is MRS. ### Claim 13 (depends on 1) The method of claim 1, wherein said growth substrate comprises a growth medium. ### Claim 14 (depends on 13) The method of claim 13, wherein said growth medium is selected from the group consisting of LB, LBGM, milk and MRS. ### Claim 15 (depends on 1) The method of claim 1, wherein when said biofilm-producing bacteria are of the Bacillus genus and said beneficial bacteria are of the Lactobacillales order, said growth substrate is LBGM, milk or MRS. ### Claim 16 (depends on 15) The method of claim 15, wherein said conditions comprise a pH of about 6.5-8. ### Claim 17 (depends on 16) The method of claim 16, wherein said conditions comprise a pH of 6.8-7.5. ### Claim 18 (depends on 1) The method of claim 1 wherein said growth substrate comprises acetoin. ### Claim 19 (depends on 1) The method of claim 1, further comprising dehydrating said biofilm following said isolating. ### Claim 20 (depends on 1) The method of claim 1, wherein said beneficial bacteria comprise no more than 50 bacterial species. ### Claim 21 (depends on 1) The method of claim 1, wherein said biofilm-producing bacteria are a single species of biofilm-producing bacteria.