(12) United States Patent (10) Patent No.: US 7,645,597 B2 Metz Et Al

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(12) United States Patent (10) Patent No.: US 7,645,597 B2 Metz Et Al USOO7645597B2 (12) United States Patent (10) Patent No.: US 7,645,597 B2 Metz et al. (45) Date of Patent: Jan. 12, 2010 (54) PUFA POLYKETIDE SYNTHASE SYSTEMS CI2N 5/82 (2006.01) AND USES THEREOF CI2P 7/64 (2006.01) (52) U.S. Cl. ....................... 435/134:435/419,435/189: (75) Inventors: James G. Metz, Longmont, CO (US); 435/252.3; 435/320.1; 435/410,536/23.2 Craig A. Weaver, Boulder, CO (US); (58) Field of Classification Search ....................... None William R. Barclay, Boulder, CO (US); See application file for complete search history. James H. Flatt, Colorado Springs, CO (US) (56) References Cited (73) Assignee: Martek Biosciences Corporation, U.S. PATENT DOCUMENTS Columbia, MD (US) 5,130,242 A 7/1992 Barclay et al. (*) Notice: Subject to any disclaimer, the term of this inued patent is extended or adjusted under 35 (Continued) U.S.C. 154(b) by 0 days. FOREIGN PATENT DOCUMENTS (21) Appl. No.: 11/778,608 CA 252O795 10, 2004 (22) Filed: Jul. 16, 2007 (Continued) (65) Prior Publication Data OTHER PUBLICATIONS US 2008/OO44871 A1 Feb. 21, 2008 Abbadi et al., Eur, J. Lipid Sci. Technol. 103: 106-113 (2001). Related U.S. Application Data (Continued) (60) Division of application No. 1 1/676.971, filed on Feb. Primary Examiner Anand U Desai 20, 2007, which is a division of application No. Assistant Examiner William W. Moore 10/810,352, filed on Mar. 26, 2004, now Pat. No. (74) Attorney, Agent, or Firm—Sheridan Ross P.C. 7.211,418, and a continuation-in-part of application No. 10/124,800, filed on Apr. 16, 2002, now Pat. No. (57) ABSTRACT 7.247.461, and a continuation-in-part of application No. 09/231,899, filed on Jan. 14, 1999, now Pat. No. The invention generally relates to polyunsaturated fatty acid 6,566,583. (PUFA) polyketide synthase (PKS) systems, to homologues (60) Provisional application No. 60/457,979, filed on Mar thereof, to isolated nucleic acid molecules and recombinant 26, 2003, provisional appl ication No. 60,284.066. nucleic acid molecules encoding biologically active domains filed O Apr. 16, 2001, provisional application No. of Such a PUFA PKS system, to genetically modified organ 60/298.796, filed on Jun. 15, 2001, provisional appli- isms comprising PUFA PKS systems, to methods of making cation No. 60,323,269, filedons ep 18, 2001 and using Such systems for the production of bioactive mol s1- Y - s • Y-s ecules of interest, and to novel methods for identifying new (51) Int. Cl. bacterial and non-bacterial microorganisms having Such a CI2N 15/53 (2006.01) PUFA PKS system. CI2N 15/74 (2006.01) CI2N 15/79 (2006.01) 21 Claims, 3 Drawing Sheets Orf A Orf B KS CLF AT ER Orf C 166 kDa US 7,645,597 B2 Page 2 U.S. PATENT DOCUMENTS WO WO 2004/087879 10, 2004 WO WO 2006/008099 1, 2006 5,246,841 A 9, 1993 Yazawa et al. WO WO 2006/034228 3, 2006 5,310,242 A 5, 1994 Golder 5,639,790 A 6, 1997 Voelker et al. OTHER PUBLICATIONS 5,672.491 A 9, 1997 Khosia et al. 5,683,898 A 11/1997 Yazawa et al. Allen et al., Appl. Envir. Microbiol. 65(4): 1710-1720 (1999). 5,798,259 A 8, 1998 Yazawa et al. Bateman et al., Nucl. Acids Res., 30(1):276-280 (2002). 5,908,622 A 6/1999 Barclay Bentley et al., Annu. Rev. Microbiol., 53:411-46 (1999). 6,033,883. A 3, 2000 Barr et al. Bisang et al., Nature, 401:502-505 (1999). 6,140,486 A 10, 2000 Facciotti et al. Bork, TIG, 12(10):425-427 (1996). 6,503,706 B1 1/2003. Abken et al. Brenner, TIG, 15(4): 132-133 (1999). 6,566,583 B1 5, 2003 Facciotti et al. Broun et al., Science, 282:1315-1317 (1998). 7,001,772 B2 2/2006 Roessler et al. 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