( 12 ) United States Patent

( 12 ) United States Patent

US010167477B2 (12 ) United States Patent ( 10 ) Patent No. : US 10 , 167, 477 B2 Pharkya (45 ) Date of Patent : Jan . 1 , 2019 ( 54 ) MICROORGANISMS AND METHODS FOR 5 , 143, 834 A 9 / 1992 Glassner et al . 5 , 168 ,055 A 12 / 1992 Datta et al. THE PRODUCTION OF ANILINE 5 , 168 ,056 A 12 / 1992 Frost .. .. 435 / 472 5 , 173 ,429 A 12/ 1992 Gaddy et al. (71 ) Applicant: Genomatica , Inc. , San Diego , CA (US ) 5 , 182 , 199 A 1 / 1993 Hartley 5 , 192 ,673 A 3 / 1993 Jain et al . (72 ) Inventor : Priti Pharkya, San Diego , CA (US ) 5 , 403 , 721 A 4 / 1995 Ward , Jr . et al. 5 ,413 , 922 A 5 / 1995 Matsuyama et al. 5 ,416 , 020 A 5 / 1995 Severson et al . (73 ) Assignee : GENOMATICA , INC . , San Diego, CA 5 ,457 , 040 A 10 / 1995 Jarry et al . (US ) 5 ,478 , 952 A 12 / 1995 Schwartz 5 ,487 ,987 A 1 / 1996 Frost et al. ( * ) Notice : Subject to any disclaimer, the term of this 5 , 504 ,004 A 4 / 1996 Guettler et al. patent is extended or adjusted under 35 5 , 521 ,075 A 5 / 1996 Guettler et al . U . S . C . 154 ( b ) by 0 days. 5 ,573 , 931 A 11/ 1996 Guettler et al. 5 ,616 , 496 A 4 / 1997 Frost et al. 5 ,686 ,276 A 11 / 1997 Lafend et al . ( 21 ) Appl. No. : 15 /914 , 308 5 ,700 , 934 A 12 / 1997 Wolters et al. 5 ,770 ,435 A 6 / 1998 Donnelly et al . (22 ) Filed : Mar. 7 , 2018 5 , 807 , 722 A 9 / 1998 Gaddy et al . 5 ,869 , 301 A 2 / 1999 Nghiem et al. (65 ) Prior Publication Data 5 , 908 , 924 A 6 / 1999 Burdette et al. 5 , 958 ,745 A 9 / 1999 Gruys et al . US 2018 /0195076 A1 Jul . 12 , 2018 6 , 117 ,658 A 1 / 2000 Dennis et al. 6 , 133 ,014 A 10 /2000 Mukouyama et al . Related U . S . Application Data 6 , 136 , 577 A 10 / 2000 Gaddy et al. (60 ) Division of application No . 15 / 141, 002 , filed on Apr. (Continued ) 28 , 2016 , now abandoned , which is a continuation of FOREIGN PATENT DOCUMENTS application No . 14 /664 , 033, filed on Mar. 20 , 2015 , now abandoned , which is a continuation of CN 1 358 841 7 /2002 application No . 12 / 910 ,671 , filed on Oct . 22 , 2010 , EP 0 494 078 7 / 1992 now abandoned . (Continued ) (60 ) Provisional application No. 61 /254 ,630 , filed on Oct. 23 , 2009 . OTHER PUBLICATIONS Abadjieva et al. , “ The Yeast ARG7Gene Product is Autoproteolyzed (51 ) Int . Ci. to Two Subunit Peptides , Yielding Active Ornithine Acetyltransferase ." C12P 13 /00 ( 2006 .01 ) J . Biol. Chem . 275 ( 15 ) :11361 - 11367 ( 2000 ) . C12N 15 / 52 ( 2006 . 01 ) Abe et al ., “ Discovery of amide (peptide ) bond synthetic activity in C12N 15 / 70 ( 2006 .01 ) Acyl - CoA synthetase , " J . Biol . Chem , 283 ( 17 ) :11312 -11321 (2008 ) . C12N 9 /88 ( 2006 .01 ) Aberhart and Hsu , “ Stereospecific hydrogen loss in the conversion C12N 9 / 10 ( 2006 .01 ) of ( H , isobutyrate to B -hydroxyisobutyrate in Pseudomonas putida . (52 ) U . S . CI. The stereochemistry of hydroxyisobutyrate dehydrogenase. ” J . Chem . CPC . .. .. .. C12N 15 /52 (2013 . 01 ) ; C12N 9 / 1096 Soc . rPerkin116 : 1404 - 1406 ( 1979 ) . ( 2013 .01 ) ; C12N 9 / 88 (2013 .01 ) ; C12N 15 / 70 Abiko et al ., “ Localization of NAD - isocitrate dehydrogenase and (2013 .01 ) ; C12P 13 /001 ( 2013. 01 ) ; C12Y glutamate dehydrogenase in rice roots : candidates for providing 206 /01085 ( 2013 .01 ) ; C12Y 401/ 01024 carbon skeletons to NADH - glutamate synthase , ” Plant Cell Physiol. (2013 .01 ) ; C12Y 401/ 03038 (2013 .01 ) 46 : 1724 - 1734 (2005 ) . (58 ) Field of Classification Search (Continued ) CPC . .. C12Y 206 /01085 ; C12N 9 / 1096 ; C12P 13 /001 Primary Examiner — Delia M Ramirez See application file for complete search history . ( 74 ) Attorney , Agent, or Firm — Kagan Binder , PLLC (56 ) References Cited (57 ) ABSTRACT A non - naturally occurring microbial organism having an U . S . PATENT DOCUMENTS aniline pathway includes at least one exogenous nucleic acid 3 ,513 , 209 A 5 / 1970 Clement encoding an aniline pathway enzyme expressed in a suffi 3 ,912 ,586 A 10 / 1975 Kaneyuki et al. cient amount to produce aniline . The aniline pathway 3 , 965 , 182 A 6 / 1976 Worrel 4 , 048 , 196 A 9 / 1977 Broecker et al. includes ( 1 ) an aminodeoxychorismate synthase, an amin 4 ,082 , 788 A 4 / 1978 Mims odeoxychorismate lyase , and a 4 - aminobenzoate carboxy 4 , 190 , 495 A 2 / 1980 Curtiss lyase or ( 2 ) an anthranilate synthase and an anthranilate 4 , 301 , 077 A 11 / 1981 Pesa et al. decarboxylase . A method for producing aniline , includes 4 , 371 ,614 A 2 / 1983 Anderson et al . .. .. .. 435 / 108 culturing these non -naturally occurring microbial organisms 4 ,624 , 920 A 12 / 1986 Inoue 4 ,652 ,685 A 3 / 1987 Cawse et al. under conditions and for a sufficient period of time to 4 , 871 ,667 A 10 / 1989 Imada et al. produce aniline . 5 , 079 , 143 A 1 / 1992 Klein et al . 5 , 143 , 833 A 9 / 1992 Datta 8 Claims, 2 Drawing Sheets US 10 ,167 ,477 B2 Page 2 (56 ) References Cited 2009 / 0075351 A1 3 / 2009 Burk et al . 2009 / 0305364 A112 / 2009 Burgard et al . U . S . PATENT DOCUMENTS 2010 /0009419 AL 1 / 2010 Burk et al. 2010 / 0099925 Al 4 / 2010 Kharas 6 , 159 ,738 A 12/ 2000 Donnelly et al . 6 , 194 , 572 B1 2 / 2001 Buijs et al . FOREIGN PATENT DOCUMENTS 6 ,214 , 592 B1 4 / 2001 Crouzet et al. 6 , 274 ,790 B1 8 / 2001 Kunst et al . EP 1 473 368 11 /2004 6 , 280 , 986 B1 8 / 2001 Hespell et al. EP 2017 344 1 / 2009 RE37 , 393 E 9 / 2001 Donnelly et al. 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