Acetyl-CoA carboxylase variants
The disclosure relates to acetyl-CoA carboxylase (ACC) variants and host cells expressing them for the production of malonyl-CoA derived compounds including fatty acid derivatives. Further contemplated are methods of producing increased amounts of malonyl-CoA derived compounds and related cell cultures.
1. A variant operon comprising a genetically modified accBC promoter that controls expression of a biotin carboxyl carrier protein (BCCP), wherein the promoter is selected from any one of SEQ ID NOS: 93, 94, 95, and 96, wherein the operon results in an increase in BCCP expression in a recombinant microbial cell as compared to a corresponding wild type microbial cell, and wherein the operon confers to the recombinant microbial cell improved production of a malonyl-CoA-derived compound when compared to the corresponding wild type microbial cell.
2. The variant operon of claim 1 , wherein the malonyl-CoA-derived compound is a fatty acid derivative selected from the group consisting of a fatty acid, a fatty acid methyl ester (FAME), a fatty acid ethyl ester (FAEE), a fatty alcohol, a fatty amine, a beta hydroxy fatty acid derivative, a bifunctional fatty acid derivative, and an unsaturated fatty acid derivative.
3. The variant operon of claim 2 , wherein the malonyl-CoA-derived compound is FAME.
4. A recombinant microorganism comprising the variant operon of claim 1 .
5. A method for producing a malonyl-CoA-derived compound, the method comprising culturing a recombinant microorganism comprising: (a) a variant biotin carboxyl carrier protein (BCCP) comprising at least one mutation in its amino acid sequence, wherein the variant BCCP comprises a polypeptide sequence selected from the group consisting of SEQ ID NOS: 4, 6, 10, 14, 16, 20, 24, 32, 48, and 70, and wherein expression of the variant BCCP confers to a recombinant cell an increased production of a malonyl-CoA-derived compound when compared to a corresponding wild type cell; and (b) the variant operon of claim 1 , in a fermentation broth containing a carbon source.
6. A method of producing a malonyl-CoA-derived compound, comprising culturing a recombinant microorganism in a fermentation broth containing a carbon source, wherein the recombinant microorganism comprises the variant operon of claim 1 .
7. The method of claim 6 , wherein the malonyl-CoA-derived compound is a fatty acid derivative selected from the group consisting of a fatty acid, a fatty acid methyl ester (FAME), a fatty acid ethyl ester (FAEE), a fatty alcohol, a fatty amine, a beta hydroxy fatty acid derivative, a bifunctional fatty acid derivative, and an unsaturated fatty acid derivative.
8. The method of claim 7 , wherein the malonyl-CoA-derived compound is FAME.
9. A recombinant microorganism having increased expression of a nucleic acid sequence comprising accB or accC or a combination thereof, wherein the increased expression is due to one or more genetically modified accBC promoters that drive expression of the nucleic acid sequence, wherein the genetically modified promoter is selected from any one of SEQ ID NOS: 93, 94, 95, and 96, and wherein the increased expression results in an increased production of a malonyl-CoA-derived compound when the microorganism is cultured with a carbon source.
10. The recombinant microorganism of claim 9 , wherein the malonyl-CoA-derived compound is selected from the group consisting of a fatty acid, a fatty acid methyl ester (FAME), a fatty acid ethyl ester (FAEE), a fatty alcohol, a fatty amine, a beta hydroxy fatty acid derivative, a bifunctional fatty acid derivative, and an unsaturated fatty acid derivative.
11. The recombinant microorganism of claim 10 , wherein the malonyl-CoA-derived compound is FAME.
12. The recombinant microorganism of claim 9 , wherein the microorganism is selected from the group consisting of Escherichia, Bacillus, Cyanophyta, Lactobacillus, Zymomonas, Rhodococcus, Pseudomonas, Aspergillus, Trichoderma, Neurospora, Fusarium, Humicola, Rhizomucor, Kluyveromyces, Pichia, Mucor, Myceliophtora, Penicillium, Phanerochaete, Pleurotus, Trametes, Chrysosporium, Saccharomyces, Stenotrophamonas, Schizosaccharomyces, Yarrowia , and Streptomyces.
13. The microorganism of claim 12 , wherein (a) the Escherichia is Escherichia coli ; and/or (b) the Cyanophyta is selected from the group consisting of Prochiorococcus, Synechococcus, Synechocystis, Cyanothece , and Nostoc punctiforme.
14. The microorganism of claim 13 , wherein the Cyanophyta is selected from the group consisting of Synechococcus elongates PCC7942, Synechocystis sp. PCC6803, and Synechococcus sp. PCC7001.
15. A method of producing a malonyl-CoA-derived compound, comprising culturing a microorganism in a fermentation broth containing a carbon source, wherein the microorganism is a microorganism of claim 9 .