Microorganisms and methods for the production of caprolactone

1. A non-naturally occurring microbial organism, said microbial organism having a caprolactone pathway and comprising heterologous nucleic acids 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 1I;

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

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

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

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

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

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

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

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

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

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

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

(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.

2. The non-naturally occurring microbial organism of claim 1 , wherein said microbial organism comprises two, three, four, five, six or seven heterologous nucleic acids each encoding a caprolactone pathway enzyme.

3. The non-naturally occurring microbial organism of claim 2 , wherein said microbial organism comprises heterologous nucleic acids encoding each of the enzymes of at least one of the pathways selected from (1)-(22).

4. The non-naturally occurring microbial organism of claim 1 , wherein said non-naturally occurring microbial organism is in a substantially anaerobic culture medium.

5. The non-naturally occurring microbial organism of claim 1 , further comprising:

(i) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from an ATP-citrate lyase, citrate lyase, a citryl-CoA synthetase, a citryl-CoA, a fumarate reductase, and an alpha-ketoglutarate:ferredoxin oxidoreductase;

(ii) a reductive TCA pathway comprising at least one exogenous nucleic acid encoding a reductive TCA pathway enzyme, wherein said at least one exogenous nucleic acid is selected from a pyruvate:ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a CO dehydrogenase, and an H 2 hydrogenase; or

(iii) at least one exogenous nucleic acid encoding an enzyme selected from a CO dehydrogenase, an H 2 hydrogenase, or a combination thereof.

6. The non-naturally occurring microbial organism of claim 5 , wherein said microbial organism comprising (i) further comprises an exogenous nucleic acid encoding an enzyme selected from a pyruvate:ferredoxin oxidoreductase, an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, an acetate kinase, a phosphotransacetylase, an acetyl-CoA synthetase, an NAD(P)H:ferredoxin oxidoreductase, ferredoxin, and combinations thereof.

7. The non-naturally occurring microbial organism of claim 5 , wherein said microbial organism comprising (ii) further comprises an exogenous nucleic acid encoding an enzyme selected from an aconitase, an isocitrate dehydrogenase, a succinyl-CoA synthetase, a succinyl-CoA transferase, a fumarase, a malate dehydrogenase, and combinations thereof.

8. The non-naturally occurring microbial organism of claim 5 , wherein said microbial organism comprising (i) comprises three exogenous nucleic acids encoding an ATP-citrate lyase or a citrate lyase, a fumarate reductase, and an alpha-ketoglutarate:ferredoxin oxidoreductase;

wherein said microbial organism comprising (ii) comprises five exogenous nucleic acids encoding a pyruvate:ferredoxin oxidoreductase, a phosphoenolpyruvate carboxylase, a phosphoenolpyruvate carboxykinase, a CO dehydrogenase, and an H 2 hydrogenase; or

wherein said microbial organism comprising (iii) comprises two exogenous nucleic acids encoding a CO dehydrogenase and an H 2 hydrogenase.

9. A method for producing caprolactone, comprising culturing the non-naturally occurring microbial organism of claim 1 under conditions and for a sufficient period of time to produce caprolactone.

10. Culture medium comprising bioderived caprolactone produced by a non-naturally occurring microbial organism of claim 1 .

11. The culture medium of claim 10 , wherein said culture medium is separated from a non-naturally occurring microbial organism having a caprolactone pathway.

12. Bioderived caprolactone produced by a non-naturally occurring microbial organism of claim 1 .

13. A composition comprising said bioderived caprolactone of claim 12 and a compound other than said bioderived caprolactone.

14. A biobased polymer comprising said bioderived caprolactone of claim 12 .

15. The biobased polymer of claim 14 , wherein said polymer comprises a portion of said bioderived caprolactone as a repeating unit.

16. A biobased resin comprising said bioderived caprolactone of claim 12 .

17. The non-naturally occurring microbial organism of claim 1 , wherein the caprolactone is produced at an intracellular concentration of about 0.1 mM to about 200 mM.

18. The method of claim 9 , wherein the caprolactone is produced at an intracellular concentration of about 0.1 mM to about 200 mM.

19. Bioderived caprolactone produced according to the method of claim 9 .

20. The bioderived caprolactone of claim 19 , wherein the bioderived caprolactone is separated from the culture.