Hybrid polyketide synthases
The present invention provides for a polyketide synthase (PKS) capable of synthesizing an even-chain or odd-chain diacid or lactam or diamine. The present invention also provides for a host cell comprising the PKS and when cultured produces the even-chain diacid, odd-chain diacid, or KAPA. The present invention also provides for a host cell comprising the PKS capable of synthesizing a pimelic acid or KAPA, and when cultured produces biotin.
1. At least one recombinant nucleic acid encoding-a non-naturally occurring polyketide synthase (PKS) that synthesizes pimelic acid, wherein the PKS is a hybrid PKS comprising (a) a loading module that loads malonate comprising a malonyl-specific AT domain linked to an acyl carrier protein (ACP) domain, wherein the ACP domain is linked to (b) an extender module comprising a ketosynthase (KS) domain, an acytransferase (AT) domain linked to a dehydratase (DH) domain linked to an enoyl reductase (ER) domain linked to a ketoreductase (KR) domain linked to an acyl carrier protein (ACP) domain; and (c) a further extender modulus, that extends an extended product of (b), comprising a KS domain linked to an AT domain linked to a DH domain linked to a KR domain linked to an ACP domain, wherein the extender module of (b) and the further extender molecule of (c) each extend with malonate; and wherein the extender module of (c) comprises a hydrolytic thioesterase (TE) domain at the C-terminus.
2. A replicon comprising the at least one recombinant nucleic acid of claim 1 , wherein the replicon is capable of stable maintenance in a host cell.
3. The replicon of claim 2 , wherein the replicon is a plasmid or vector.
4. The replicon of claim 3 , wherein the vector is an expression vector.
5. A host cell comprising the at least one recombinant nucleic acid of claim 1 .
6. A host cell comprising the replicon of claim 2 .
7. The host cell of claim 5 , wherein the host cell, when cultured, produces pimelic acid.
8. The host cell of claim 7 , wherein the host cell further comprises one or more nucleic acids encoding pimelyl-CoA synthetase, 8-amino-7-oxononanoate synthase, 7,8-diamino-pelargonic acid (DAPA) synthase, dethiobiotin synthase, and biotin synthase, wherein the PKS synthesizes pimelic acid, and when cultured, the host cell produces biotin.
9. A method of producing pimelic acid comprising: providing the host cell of claim 7 , and culturing said host cell in a suitable culture medium such that pimelic acid is produced.
10. The method of claim 9 , further comprising isolating pimelic acid.
11. A method of producing a biotin comprising: providing the host cell of claim 8 , and culturing said host cell in a suitable culture medium such that the biotin is produced.