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(12) United States Patent (10) Patent No.: US 9,631,211 B2 Scholten Et Al

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(12) United States Patent (10) Patent No.: US 9,631,211 B2 Scholten Et Al USOO9631211 B2 (12) United States Patent (10) Patent No.: US 9,631,211 B2 Scholten et al. (45) Date of Patent: *Apr. 25, 2017 (54) BACTERIAL STRAIN AND FERMENTATIVE 8,883,466 B2 * 1 1/2014 Scholten .............. C12N 9/OOO6 PROCESS FOR PRODUCING SUCCNC 435/145 ACD 2007/0042481 A1 2/2007 Lee et al. 2008/0293101 A1 11/2008 Peters et al. 2009/O1378.25 A1 5/2009 Bauduin et al. (71) Applicant: BASF SE, Ludwigshafen (DE) 2009. O155869 A1 6/2009 Buelter et al. 2010, 004.4626 A1 2/2010 Fischer et al. (72) Inventors: Edzard Scholten, Mannheim (DE); 2010, O159542 A1 6, 2010 Scholten et al. Dirk Dagele, Vogtsburg (DE); Stephan 2010, O159543 A1 6, 2010 Scholten et al. Haefner, Speyer (DE); Hartwig 2010/0324258 A1 12/2010 Zelder et al. Schroder, Nussloch (DE) 2011/0300.589 A1 12/2011 Schroder et al. (73) Assignee: BASF SE, Ludwigshafen (DE) FOREIGN PATENT DOCUMENTS EP O805208 A1 5, 1997 (*) Notice: Subject to any disclaimer, the term of this EP 1842843 A1 10, 2007 patent is extended or adjusted under 35 EP 2202294 A1 6, 2010 EP 2204443 A1 T 2010 U.S.C. 154(b) by 224 days. JP 200811714 1, 2008 This patent is Subject to a terminal dis WO WO-02/00846 A1 1, 2002 claimer. WO WO-03/04O690 A2 5, 2003 WO WO-2005/052135 A1 6, 2005 WO WO-2006/034156 A2 3, 2006 (21) Appl. No.: 14/045,096 WO WO-2006/066839 A2 6, 2006 WO WO-2008/013405 A1 1, 2008 (22) Filed: Oct. 3, 2013 WO WO-2009/024294 A1 2, 2009 (65) Prior Publication Data OTHER PUBLICATIONS US 2014/OO3O778 A1 Jan. 30, 2014 Loughney et al. (Nucleic Acids Res., vol. 10, No. 5, 1982, pp. 1607-1624).* Related U.S. Application Data Database Em-Pro Mannheimia succiniproducens MBEL55E Sep. (62) Division of application No. 12/673.714, filed as 18, 2004, XPO02498827 Accession No. AEO 16827 nt 1495.543 151059 nt 2233952-223546899.7% identity with Seq ID 1 abstract. application No. PCT/EP2008/0067.14 on Aug. 14, Dharmadi, Y., et al., “Anaerobic Fermentation of Glycerol by 2008, now Pat. No. 8,574,875. Escherichia coli: A New Platform for Metabolic Engineering.” (30) Foreign Application Priority Data Biotechnology and Bioengineering, vol. 94, No. 5, pp. 821-829 (2006). Hong, S.H. et al., “The genome sequence of the capnophilic rumen Aug. 17, 2007 (EP) ..................................... O7114574 bacterium Mannheimia succiniciproducens' Nature Biotechnology, vol. 22, No. 10, (Oct. 2004), pp. 1275-1281, XP002498825 ISSN: (51) Int. Cl. 1087-0156 table 1. CI2P I 7/10 (2006.01) Janssen, P. H., "Characterization of a Succinate-fermenting anaero CI2P 7/46 (2006.01) bic bacterium isolated from a glycolate-degrading mixed culture.” CI2N L/32 (2006.01) Arch Microbiol (1991), vol. 155: pp. 288-293. CI2R I/OI (2006.01) Lee, J., “Biological conversion of lignocellulosic biomass to etha (52) U.S. Cl. nol.” Journal of Biotechnology, vol. 56 (1997), pp. 1-24. CPC .................. CI2P 7/46 (2013.01); CI2N I/32 (2013.01): CI2R I/01 (2013.01) (Continued) (58) Field of Classification Search None Primary Examiner — Hope Robinson See application file for complete search history. (74) Attorney, Agent, or Firm — Marshall, Gerstein & Borun LLP (56) References Cited U.S. PATENT DOCUMENTS (57) ABSTRACT 4.550,185 A 10/1985 Mabry et al. 5,504,004 A 4, 1996 Guettler et al. The present invention relates to a novel bacterial strain 5,573.931 A 11/1996 Guettler et al. designated DD1, which has the ability to produce organic 5,723,322 A 3, 1998 Guettler et al. 6,596,521 B1 7/2003 Chang et al. acids, in particular Succinic acid (SA), which was originally 7,063,968 B2 6, 2006 Lee et al. isolated from bovine rumen, and is capable of utilizing 7,192,761 B2 3/2007 Zeikus et al. glycerol as a carbon Source; and variant strains derived there 7,256,016 B2 8, 2007 San et al. from retaining said capability; as well as to methods of 7,262,046 B2 8, 2007 Ka-Yiu et al. 7,470,531 B2 12/2008 Rehberger et al. producing organic acids, in particular Succinic acid by 8,673,598 B2 * 3/2014 Schroder .................. C12N 1/32 making use of said microorganism. 435/121 8,877.466 B2 * 1 1/2014 Scholten .............. C12N 9,0008 435/145 3 Claims, 9 Drawing Sheets US 9,631.211 B2 Page 2 (56) References Cited “Pyruvate formate lyase-activating enzyme 1 Shigella boydii CDC 3083-94, Database NCBI, Accession No. YP 001880903.1, Jan. 5, 2011. OTHER PUBLICATIONS “Formate acetyltransferase 1". Database UniProtKB, Accession No. Lee P.C., et al., “Isolation and characterization of a new Succinic P09373, Feb. 8, 2011. acid-producing bacterium, Mannheimia “Formate acetyltransferase 2'. Database UniProtKB, Accession No. succiniciproducensMBEL55E, from bovine rumen.” Appl P32674, Feb. 8, 2011. Microbiol Biotechnol (2002) vol. 58, pp. 663-668. "Keto-acid formate acetyltransferase', Database UniProtKB, Lee, P.C., et al. “Succinic Acid Production with Reduced By Accession No. P42632, Feb. 8, 2011. 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