Producing a trimethylpentanoic acid using hybrid polyketide synthases
The present invention provides for a polyketide synthase (PKS) capable of synthesizing trimethylpentanoic acid. The present invention also provides for a host cell comprising the PKS and when cultured produces the trimethylpentanoic acid. The present invention also provides for a method of producing the trimethylpentanoic acid, comprising: providing a host cell of the present invention, and culturing said host cell in a suitable culture medium such that the trimethylpentanoic acid is produced, optionally isolating the trimethylpentanoic acid, and optionally, reducing the isolated trimethylpentanoic acid into a trimethylpentanol or an iso-octane.
1. A method of producing a trimethylpentanoic acid, comprising: providing a prokaryotic host cell and culturing said host cell in a suitable culture medium such that the trimethylpentanoic acid is produced, wherein the host cell comprises a recombinant nucleic acid encoding a hybrid polyketide synthase (PKS) having a module from two or more PKS polypeptides, and further, wherein the hybrid PKS comprises: a loading module comprising an acyltransferase (AT) and an acyl carrier protein (ACP) that uses pivaloyl-CoA as a starter unit, and a module comprising a ketosynthase (KS), methylmalonyl-CoA acyltransferase (mmAT), dehydratase (DH), enoylreductase (ER), ketoreductase (KR), and ACP that uses 2-methylmalonyl-CoA as an extending unit; and a thioesterase (TE).
2. The method of claim 1 , further comprising isolating the trimethylpentanoic acid.
3. The method of claim 2 , further comprising reducing the trimethylpentanoic acid to produce a trimethylpentanol or an iso-octane.
4. The method of claim 1 , wherein the TE is an Ery TE.
5. The method of claim 1 , wherein the module that uses 2-methylmalonyl-CoA as an extending unit comprises KS of Ave Load-Mod1, and an mmAT, DH, ER, KR, and ACP of Ery Mod4 or Nan Mod2.
6. The method of claim 1 , wherein the loading module comprises AT L and ACP L of Ave Load-Mod1.
7. The method of claim 6 , wherein the module that uses 2-methylmalonyl-CoA as an extending unit comprises KS of Ave Load-Mod1, and an mmAT, DH, ER, KR, and ACP of Ery Mod4 or NanMod 2.
8. The method of claim 7 , wherein the TE is an Ery TE.
9. The method of claim 1 , wherein the loading module comprises AT L , ACP L and KS1 of But Load Module of Streptomyces rochei.
10. The method of claim 9 , wherein the module that uses 2-methylmalonyl-CoA as an extending unit comprises an mmAT, DH, ER, KR, ACP of Ery Mod4.
11. The method of claim 10 , wherein the TE is an Ery TE.
12. The method of claim 1 , wherein the host cell comprises a replicon comprising the recombinant nucleic acid encoding the hybrid polyketide synthase, wherein the replicon is stably maintained in the host cell.
13. The method of claim 12 , wherein the replicon is a plasmid or vector.
14. The method of claim 13 , wherein the vector is an expression vector.
15. The method of claim 1 , wherein the host cell further comprises PrpE, and PccA or AccB.
16. The method of claim 1 , wherein the host cell further comprises MCR, MSR, HPCS, HPCD, and ACR.
17. The method of claim 1 , wherein the host cell is an Escherichia coli host cell.