Hybrid polyketide synthases

Patent metadata

Field	Value
Publication number	US-11505791-B2
Application number	US-201916700845-A
Country	US
Kind code	B2
Filing date	Dec 2, 2019
Priority date	Oct 29, 2010
Publication date	Nov 22, 2022
Grant date	Nov 22, 2022

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Univ California

Inventors

• Fortman Jeffrey L
• Hagen Andrew
• Katz Leonard
• Keasling Jay D
• Poust Sean
• Zhang Jingwei
• Zotchev Sergey

Classifications

• C12P13/005

  Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids · CPC title

• C12P13/04

  Alpha- or beta- amino acids {(other amino acids C12P13/005)} · CPC title

• C12P7/44

  Polycarboxylic acids · CPC title

• C12N9/93 Primary

  Ligases (6) · CPC title

• C12P17/186

  containing a 2-oxo-thieno[3,4-d]imidazol nucleus, e.g. Biotin · CPC title