Producing alpha-olefins using polyketide synthases
The present invention provides for a polyketide synthase (PKS) capable of synthesizing an α-olefin, such as 1-hexene or butadiene. The present invention also provides for a host cell comprising the PKS and when cultured produces the α-olefin.
1. A non-naturally occurring hybrid polyketide synthase (PKS) that comprises a terminal module comprising a sulfotransferase (ST) domain having at least 70% identity to the amino acid sequence of SEQ ID NO:11; and a thioesterase (TE) domain having at least 70% identity to the amino acid sequence of SEQ ID NO:12; and a loading module from an erythromycin (DEBS) loading domain, wherein the non-naturally occurring hybrid PKS synthesizes an α-olefin.
2. A recombinant nucleic acid encoding the non-naturally occurring hybrid PKS of claim 1 .
3. A method of producing an α-olefin, comprising: providing a host cell comprising the recombinant nucleic acid of claim 2 , and culturing said host cell in a suitable culture medium such that the α-olefin is produced.
4. A non-naturally occurring hybrid PKS that synthesizes 1-butene, wherein the non-naturally occurring hybrid PKS comprises a terminal module comprising an acyltransferase (AT) domain, a ketoreductase (KR) domain, an acyl carrier protein (ACP) domain; an ST domain having at least 90% identity to the amino acid sequence of SEQ ID NO:11; and a TE domain having at least 90% identity to the amino acid sequence of SEQ ID NO: 12; and a propionyl-CoA loading module.
5. The non-naturally occurring hybrid PKS of claim 4 , wherein the terminal module comprises an AT domain, a KR domain, and an ACP domain, each of which domains is from Lyngbya majuscula CurM having the amino acid sequence of SEQ ID NO: 1; an ST domain having the amino acid sequence of SEQ ID NO: 11; and a TE domain having the amino acid sequence of SEQ ID NO: 12.
6. The non-naturally occurring hybrid PKS of claim 5 , wherein the non-naturally occurring hybrid PKS comprises the loading module for propionyl-CoA and a ketosynthase (KS) domain from an erythromycin (DEBS) loading domain.
7. The non-naturally occurring hybrid PKS of claim 4 , wherein the non-naturally occurring hybrid PKS comprises the loading module for propionyl-CoA and a ketosynthase (KS) domain from an erythromycin (DEBS) loading domain.
8. A recombinant nucleic acid encoding the non-naturally occurring hybrid PKS of claim 4 .
9. A host cell comprising the recombinant nucleic acid of claim 8 .
10. The host cell of claim 9 , wherein the host cell when cultured produces 1-butene.
11. A method of producing an α-olefin, comprising: providing the host cell of claim 9 , and culturing said host cell in a suitable culture medium such that 1-butene is produced.
12. The method of claim 11 , further comprising isolating 1-butene.