# US 9,518,279 B2 · Bacterial strain Rhodococcus aetherivorans VKM Ac-2610D producing nitrile hydratase, method of its cultivation and method for producing acrylamide > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 9,518,279 B2 - Document ID: US9518279B2 - Status: Granted - Kind: B2 - Application number: 14/140,150 - Filed: 2013-12-24 - Published: 2014-07-03 - Granted: 2016-12-13 - Earliest claimed: 2012-12-27 - Assignees: Kemira Oyj - Applicants: Kemira Oyj - Inventors: Sergey Vladimirovich Kozulin; Tatiana Nicolaevna Kozulina; Alexey Sergeevich Kozulin - Web page: https://patentgrep.com/patent/US9518279B2 ## CPC - C12N 1/20: Bacteria; Culture media therefor - C12N 1/00: Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor - C12N 1/205: Bacterial isolates - C12P 13/02: Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)} - C12Y 402/01084: Nitrile hydratase (4.2.1.84) - C12R 2001/00: Microorganisms {; Processes using microorganisms} - C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales} - 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)} - C12Y: ENZYMES - C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS ## Abstract The invention relates to a bacterial strain belonging to the genus Rhodococcus which is a producer of a nitrile hydratase. The invention also relates to a method for producing acrylamide by hydration of acrylonitrile using a biomass of the bacterial strain and to a method of culturing the bacterial strain. ## Claims ### Claim 1 A method for producing acrylamide, the method comprising hydration of acrylonitrile using a biomass of a bacterial strain belonging to the genus Rhodococcus and having a nitrile hydratase activity, wherein hydration is carried out at a working concentration of acrylonitrile which is no more than 0.5%, and the bacterial strain is Rhodococcus aetherivorans VKM Ac-2610D. ### Claim 2 (depends on 1) The method of claim 1, wherein acrylonitrile is mixed in an aqueous suspension of the biomass of the bacterial strain to form a reaction solution; and hydration is carried out so that the working concentration of acrylonitrile in the reaction solution is maintained at no more than 0.5%. ### Claim 3 (depends on 1) The method of claim 1, wherein hydration is followed by isolation of acrylamide. ### Claim 4 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is at about 100-1000 g, or 200-1000 g, or 200-800 g, or 300-600 g, or about 400-500 g, or less than 1000 g, or less than 500 g, or less than 400 g on a dry weight of strain per 1 ton of a final product. ### Claim 5 (depends on 4) The method of claim 4, wherein the final product contains acrylamide at a concentration of at least 40%; or at least 41%; or at least 42%; or at least 43%; or at least 44%; or 40-55%; or 41-55%; 42-55%; or 43-55%; or 44-55%; or at least 45%; or at least 46%; or at least 47%; or 45-55%; or 45-54%; or 45-53%; or 45-52%; or 45-51%; or 45-50%; or 45-49%; or 46-55%; or 46-54%; or 46-53%; or 46-52%; or 46-51%; or 46-50%; or 46-49%; or 47-55%; or 47-54%; or 47-53%; or 47-52%; or 47-51%; or 47-50%; or 47-49%. ### Claim 6 (depends on 1) The method of claim 1, wherein hydration of acrylonitrile is carried out at a temperature of 10-23° C. ### Claim 7 (depends on 1) The method of claim 1, wherein hydration of acrylonitrile is carried out at a pH of 6.8-8.4. ### Claim 8 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is at about 100-1000 g on a dry weight of strain per 1 ton of a final product. ### Claim 9 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is at about 200-1000 g on a dry weight of strain per 1 ton of a final product. ### Claim 10 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is at about 200-800 g on a dry weight of strain per 1 ton of a final product. ### Claim 11 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is at about 300-600 g on a dry weight of strain per 1 ton of a final product. ### Claim 12 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is at about 400-500 g on a dry weight of strain per 1 ton of a final product. ### Claim 13 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is less than 1000 g on a dry weight of strain per 1 ton of a final product. ### Claim 14 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is less than 500 g on a dry weight of strain per 1 ton of a final product. ### Claim 15 (depends on 1) The method of claim 1, wherein the biomass of the bacterial strain is less than 400 g on a dry weight of strain per 1 ton of a final product. ### Claim 16 (depends on 4) The method of claim 4, wherein the final product contains acrylamide at a concentration of 47-55%. ### Claim 17 (depends on 4) The method of claim 4, wherein the final product contains acrylamide at a concentration of 47-54%. ### Claim 18 (depends on 4) The method of claim 4, wherein the final product contains acrylamide at a concentration of 47-52%. ### Claim 19 (depends on 4) The method of claim 4, wherein the final product contains acrylamide at a concentration of 47-52%. ### Claim 20 (depends on 4) The method of claim 4, wherein the final product contains acrylamide at a concentration of 47-49%.