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(12) United States Patent (10) Patent No.: US 9,017,983 B2 Burgard Et Al

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(12) United States Patent (10) Patent No.: US 9,017,983 B2 Burgard Et Al US009017983B2 (12) United States Patent (10) Patent No.: US 9,017,983 B2 Burgard et al. (45) Date of Patent: Apr. 28, 2015 (54) ORGANISMS FOR THE PRODUCTION OF 5,521,075 A 5/1996 Guettler et al. 13-BUTANEDOL 5,573.931 A 11/1996 Guettler et al. 9 5,616,496 A 4/1997 Frost et al. 5,661,026 A 8/1997 Peoples et al. (75) Inventors: Anthony P. Burgard, Bellefonte, PA 5,686,276 A 1 1/1997 Lafend et al. (US); Mark J. Burk, San Diego, CA 5,700.934. A 12/1997 Wolters et al. (US); Robin E. Osterhout, San Diego, 5,770.435 A 6/1998 Donnelly et al. t sS); Priti Pharkya, San Diego, CA (Continued) FOREIGN PATENT DOCUMENTS (73) Assignee: Genomatica, Inc., San Diego, CA (US) CN 1358 841 T 2002 (*) Notice: Subject to any disclaimer, the term of this EP O 494 O78 7, 1992 patent is extended or adjusted under 35 E. s 29: U.S.C. 154(b) by 654 days. GB 1230276 4f1971 GB 1314126 4f1973 (21) Appl. No.: 12/772,114 GB 1344557 1, 1974 GB 1512751 6, 1978 (22) Filed: Apr. 30, 2010 JP 50 OO6776 1, 1975 WO WO 82,03854 11, 1982 (65) Prior Publication Data (Continued) US 2010/033.0635A1 Dec. 30, 2010 OTHER PUBLICATIONS Related U.S. Application Data Abadjieva et al., “The Yeast ARG7 Gene Product is Autoproteolyzed (60) Provisional application No. 61/174,473, filed on Apr. to Two Subunit Peptides, Yielding Active Ornithine 30, 2009 Acetyltransferase.” J. Biol. Chem. 275(15): 11361-11367 (2000). (Continued) (51) Int. Cl. 2. E.2 28:8 Primary Examiner — Tekchand Saidha CI2N 9/04 3:08: Assistant Examiner — Md. Younus Meah (52) U.S. Cl (74) Attorney, Agent, or Firm — Jones Day CPC. CI2PC12Nooooo 7/18 (2013.01): (2013.01). CI2N C1215/52 (2013.01);ioioi157 (7) ABSTRACT (2013.01) A non-naturally occurring microbial organism includes a (58) Field of Classification Search microbial organism having a 1,3-butanediol (1,3-BDO) path None way having at least one exogenous nucleic acid encoding a See application file for complete search history. 1,3-BDO pathway enzyme expressed in a sufficient amount to produce 1,3-BDO. The pathway includes an enzyme selected (56) References Cited from a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP dehydrogenase, a 2-amino-4-hydroxypentanoate ami U.S. PATENT DOCUMENTS notransferase, a 2-amino-4-hydroxypentanoate oxidoreduc 3,513,209 A 5, 1970 Clement tase (deaminating), a 2-oxo-4-hydroxypentanoate decar 3,912,586 A 10/1975 Kaneyuki et al. boxylase, a 3-hydroxybutyraldehyde reductase, an AKP 3,965, 182 A 6, 1976 Worrel 4,048,196 A 9, 1977 Broecker et al. aminotransferase, an AKP oxidoreductase (deaminating), a 4,082,788 A 4, 1978 Mims 2,4-dioxopentanoate decarboxylase, a 3-oxobutyraldehyde 4,190.495 A 2f1980 Curtiss reductase (ketone reducing), a 3-oxobutyraldehyde reductase 4,301,077 A 11, 1981 Pesa et al. (aldehyde reducing), a 4-hydroxy-2-butanone reductase, an 4,624,920 A 11, 1986 Inoue 4,652,685 A 3, 1987 Cawse et al. 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A method 5,457,040 A 10/1995 Jarry et al. for producing 1,3-BDO, includes culturing such microbial 5,478,952 A 12/1995 Schwartz 5,487.987 A 1/1996 Frost et al. organisms under conditions and for a sufficient period of time 5,504,004 A 4, 1996 Guettler et al. to produce 1,3-BDO. 5,512.465 A 4, 1996 Matsuyama et al. 5,512,669 A 4/1996 Sinskey et al. 5 Claims, 7 Drawing Sheets US 9,017,983 B2 Page 2 (56) References Cited 2007/0042476 A1 2, 2007 Lee et al. 2007/OO72279 A1 3, 2007 Meynial-Salles et al. 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