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(12) United States Patent (10) Patent No.: US 7,799,545 B2 Burgard Et Al US007799545B2 (12) United States Patent (10) Patent No.: US 7,799,545 B2 Burgard et al. (45) Date of Patent: Sep. 21, 2010 (54) MICROORGANISMS FOR THE 4,082,788 A 4/1978 Mims PRODUCTION OF ADPCACID AND OTHER 4,190.495 A 2/1980 Curtiss COMPOUNDS 4,301,077 A 1 1/1981 Pesa et al. 4,624,920 A 11/1986 Inoue et al. (75)75 Inventors: Anthony P. Burgard, Bellefonte, PA 4,871,6674,652,685 A 10/19893, 1987 ImadaCawse etet al.al. (US); Priti Pharkya, San Diego, CA (US); Robin E. Osterhout, San Diego, (Continued) CA (US) FOREIGN PATENT DOCUMENTS (73) Assignee: Genomatica, Inc., San Diego, CA (US) CN 1358 841 T 2002 *) NotOt1Ce: Subjubject to anyy d1Sclaimer,disclai theh term off thithis (Continued)ontinue patent is extended or adjusted under 35 OTHER PUBLICATIONS U.S.C. 154(b) by 47 days. Alhapel et al., “Molecular and functional analysis of nicotinate (21) Appl. No.: 12/413,355 catabolism in Eubacterium barkeri,’ Proc. Natl. Acad. Sci. USA 103(33):12341-12346 (2006). (22) Filed: Mar. 27, 2009 (Continued) (65) Prior Publication Data Primary Examiner Nashaat T Nashed US 2009/0305364 A1 Dec. 10, 2009 (74) Attorney, Agent, or Firm McDermott Will & Emery • ws LLP Related U.S. Application Data (57) ABSTRACT (60) Provisional application No. 61/040,059, filed on Mar. 27, 2008. The invention provides a non-naturally occurring microbial organism having an adipate, 6-aminocaproic acid or capro (51) Int. Cl. lactam pathway. The microbial organism contains at least one CI2P 7/40 (2006.01) exogenous nucleic acid encoding an enzyme in the respective CI2N L/20 (2006.01) adipate, 6-aminocaproic acid or caprolactam pathway. The (52) U.S. Cl. ................ 435/136: 435/252.3:435/252.33 invention additionally provides a method for producing adi (58) Field of Classification Search ....................... None pate, 6-aminocaproic acid or caprolactam. The method can See application file for complete search history. include culturing an adipate, 6-aminocaproic acid or capro lactam producing microbial organism, where the microbial (56) References Cited organism expresses at least one exogenous nucleic acid encoding an adipate, 6-aminocaproic acid or caprolactam U.S. PATENT DOCUMENTS pathway enzyme in a sufficient amount to produce the respec 3,513,209 A 5, 1970 Clement tive product, under conditions and for a sufficient period of 3,912,586 A 10/1975 Kaneyuki et al. time to produce adipate, 6-aminocaproic acid or caprolactam. 3,965, 182 A 6, 1976 Worrel 4,048,196 A 9, 1977 Broecker et al. 14 Claims, 9 Drawing Sheets } Adiate HO-----o- {} Atipate-caliase k a ..., avir. PP Adipyl-CoA --- scoA &city:-&c., a defydrogenasem A. 39 s w S-(arboxy-2-pentenoyl-CoA HO---- S-3c)a Enoyl-CoA hydratase / 32. 3-ydroxyadipyl-CoA "yr-Yss-CoAHg O 3-isdiriyyacyl-CoA deihydrogenase N. : s: K NA 3-(xeadiyi-CoAw or 'scsC. Y - 3 a. 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