Acetyl-CoA carboxylases

• US 10,155,937 B2
• Filed: 09/19/2016
• Issued: 12/18/2018
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Cargill Incorporated
• Original Assignee: Cargill Incorporated
• Inventors: Liao, Hans, Mercogliano, Christopher Patrick, Wolter, Travis Robert, Louie, Michael Tai Man, Ribble, Wendy Kathleen, Lipscomb, Tanya E. W., Spindler, Eileen Colie, Lynch, Michael D.
• Primary Examiner(s): NOAKES, SUZANNE MARIE
• Application Number:
  US15/269,382
• Publication Number:
  US 20170101638A1
• Time in Patent Office: 820 Days
• Field of Search: None
• CPC Class Codes: C07C 59/01 Saturated compounds having only one carboxyl group and containing hydroxy or O-metal groups
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/0016 with NAD or NADP as acceptor (1.4.1)
• CPC Class Codes: C12N 9/93 Ligases (6)
• CPC Class Codes: C12P 7/42 Hydroxy-carboxylic acids
• CPC Class Codes: C12P 7/52 Propionic acid; Butyric acids
• CPC Class Codes: C12Y 604/01002 Acetyl-CoA carboxylase (6.4.1.2)
• CPC Class Codes: Y02P 20/52 using catalysts, e.g. selective catalysts

Abstract

The present invention provides various combinations of genetic modifications to a transformed host cell that provide increase conversion of carbon to a chemical product. The present invention also provides methods of fermentation and methods of making various chemical products.

Claims

1. A genetically modified microorganism comprising a heterologous gene encoding an acetyl-CoA carboxylase (ACCase) polypeptide fusion wherein the polypeptide fusion comprises at least two subunits of an ACCase enzyme selected from the group consisting of accA-accB, accA-accC, accA-accD, accB-accC, accB-accD, accC-accD, accA-accB-accC, accA-accB-accD, accA-accC-accD, accB-accC-accD and accA-accB-accC-accD, wherein the genetically modified microorganism further comprises a genetic modification to a nucleotide encoding a functional D-lactate dehydrogenase, pyruvate formate-lyase, phosphate acetyltransferase, pyruvate oxidase, methylglyoxal synthase, acetate kinase, phosphotransacetylase-acetate kinase, or any combination thereof.

2. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for D-lactate dehydrogenase comprises IdhA from E. coli.

3. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for pyruvate formate-lyase comprises pflB from E. coli.

4. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for phosphate acetyltransferase comprises pta from E. coli.

5. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for the pyruvate oxidase comprises poxB from E. coli.

6. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for methylglyoxal synthase comprises mgsA from E. coli.

7. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for acetate kinase comprises ackA from E. coli.

8. The genetically modified microorganism of claim 1, wherein the nucleotide encoding for phosphotransacetylase-acetate kinase comprises pta-ack from E. coli.

9. A genetically modified microorganism comprising a heterologous gene encoding an acetyl-CoA carboxylase (ACCase) polypeptide fusion wherein the polypeptide fusion comprises at least two subunits of an ACCase enzyme selected from the group consisting of accA-accB, accA-accC, accA-accD, accB-accC, accB-accD, accC-accD, accA-accB-accC, accA-accB-accD, accA-accC-accD, accB-accC-accD and accA-accB-accC-accD, wherein the genetically modified microorganism is capable of producing a C4 to C18 fatty acid chain product.

10. The microorganism of claim 9, wherein the fatty acid chain product is selected from the group consisting of fatty acid methyl esters and fatty acids.