	US 11,708,592 B2
	Microorganisms and methods for the production of caprolactone
Anthony P. Burgard, Elizabeth, PA (US); Robin E. Osterhout, San Diego, CA (US); Priti Pharkya, San Diego, CA (US); and Mark J. Burk, San Diego, CA (US)
Assigned to GENOMATICA, INC., San Diego, CA (US)
Filed by Genomatica, Inc., San Diego, CA (US)
Filed on Dec. 11, 2020, as Appl. No. 17/120,005.
Application 17/120,005 is a continuation of application No. 16/433,804, filed on Jun. 6, 2019, abandoned.
Application 16/433,804 is a continuation of application No. 15/634,726, filed on Jun. 27, 2017, granted, now 10,351,887, issued on Jul. 16, 2019.
Application 15/634,726 is a continuation of application No. 14/995,069, filed on Jan. 13, 2016, granted, now 9,719,118, issued on Aug. 1, 2017.
Application 14/995,069 is a continuation of application No. 14/596,072, filed on Jan. 13, 2015, granted, now 9,267,162, issued on Feb. 23, 2016.
Application 14/596,072 is a continuation of application No. 13/668,117, filed on Nov. 2, 2012, granted, now 8,940,509, issued on Jan. 27, 2015.
Claims priority of provisional application 61/554,920, filed on Nov. 2, 2011.
Prior Publication US 2021/0363556 A1, Nov. 25, 2021
Int. Cl. C12P 17/08 (2006.01); C12N 15/70 (2006.01); C08G 63/08 (2006.01); C07D 313/04 (2006.01); C12N 15/63 (2006.01); C12P 17/02 (2006.01); C08G 18/42 (2006.01); C12N 5/00 (2006.01); C12N 15/52 (2006.01)

CPC C12P 17/08 (2013.01) [C07D 313/04 (2013.01); C08G 18/4277 (2013.01); C08G 63/08 (2013.01); C12N 5/0018 (2013.01); C12N 15/52 (2013.01); C12N 15/63 (2013.01); C12N 15/70 (2013.01); C12P 17/02 (2013.01)]

20 Claims

1. A non-naturally occurring microbial organism, said microbial organism having a caprolactone pathway and a reductive TCA pathway, wherein said non-naturally occurring microbial organism comprises at least one exogenous nucleic acid encoding a caprolactone pathway enzyme expressed in a sufficient amount to produce caprolactone, wherein said caprolactone pathway comprises a pathway selected from:

(1) 1D, 1A, 1B, and 1C;

(2) 1E, 1B, 1C and 1D;

(3) 1F, 1A, 1B, 1C and 1D;

(4) 1F, 1E, 1B, 1C and 1D;

(5) 1A, 1B and 1G;

(6) 1E, 1B and 1G;

(7) 1F, 1A, 1B and 1G;

(8) 1F, 1E, 1B and 1G;

(9) 1A, 1B, 1C, 1J and 11;

(10) 1E, 1B, 1C, 1J and 11;

(11) 1F, 1A, 1B, 1C, 1J and 11;

(12) 1F, 1E, 1B, 1C, 1J and 11;

(13) 1A, 1B, 1H and 11;

(14) 1E, 1B, 1H and 11;

(15) 1F, 1A, 1B, 1H and 11;

(16) 1F, 1E, 1B, 1H and 11;

(17) 2A, 2B, 2C, 2D, 2E and 2F;

(18) 2A, 2B, 2C, 2D and 2G;

(19) 2A, 2B, 2C, 2D, 2E, 2J and 21;

(20) 2A, 2B, 2C, 2D, 2H and 21;

(21) 4A, 4B, 4C, 4D and 3A; and

(22) 5A, 5B, 5C an 3A,

wherein 1A is an adipyl-COA reductase, wherein 1B is an adipate semialdehyde reductase, wherein 1C is a 6-hydroxyhexanoyl-CoA transferase or a 6-hydroxyhexanoyl-CoA synthetase, wherein 1D is a 6-hydroxyhexanoyl-CoA cyclase or spontaneous cyclization, wherein 1E is an adipate reductase, wherein 1F is an adipyl-CoA transferase, an adipyl-CoA synthetase or an adipyl-CoA hydrolase, wherein 1G is a 6-hydroxyhexanoate cyclase, wherein 1H is a 6-hydroxyhexanoate kinase, wherein 1I is a 6-hydroxyhexanoyl phosphate cyclase or spontaneous cyclization, wherein 1J is a phosphotrans-6-hydroxyhexanoylase, wherein 2A is a 4-hydroxybutyryl-CoA:acetyl-CoA acyltransferase, wherein 2B is a 3-oxo-6-hydroxyhexanoyl-COA reductase, wherein 2C is a 3,6-dihydroxyhexanoyl-CoA dehydratase, wherein 2D is a 6-hydroxyhex-2-enoyl-COA reductase, wherein 2E is a 6-hydroxyhexanoyl-CoA transferase, a 6-hydroxyhexanoyl-CoA synthetase or a 6-hydroxyhexanoyl-CoA hydrolase, wherein 2F is a 6-hydroxyhexanoate cyclase, wherein 2G is a 6-hydroxyhexanoyl-CoA cyclase or spontaneous cyclization, wherein 2H is a phosphotrans-6-hydroxyhexanoylase, wherein 2I is a 6-hydroxyhexanoyl phosphate cyclase or spontaneous cyclization, wherein 2J is a 6-hydroxyhexanoate kinase, wherein 3A is a cyclohexanone monooxygenase, wherein 4A is an adipate semialdehyde dehydratase, wherein 4B is a cyclohexane-1,2-dione reductase, wherein 4C is a 2-hydroxycyclohexanone reductase, wherein 4D is a cyclohexane-1,2-diol dehydratase, wherein 5A is a 2-ketocyclohexane-1-carboxoyl-CoA hydrolase (acting on C—C), wherein 5B is a 2-ketocyclohexane-1-carboxoyl-CoA transferase, a 2-ketocyclohexane-1-carboxoyl-CoA synthetase or a 2-ketocyclohexane-1-carboxoyl-CoA hydrolase, and wherein 5C is a 2-ketocyclohexane-1-carboxylate decarboxylase.