Methods And Compositions For Improved Production Of Fatty Acids And Derivatives Thereof
The invention relates to compositions and methods, including polynucleotide sequences, amino acid sequences, and engineered host cells for producing fatty acids and derivates of fatty acids such as acyl-CoA, terminal olefins, fatty aldehydes, fatty alcohols, alkanes, alkenes, wax esters, ketones and internal olefins through altered expression of the transcription factor, fadR.
1 .- 46 . (canceled)
47 . A recombinant host cell which is genetically engineered to overexpress a FadR polypeptide and to overexpress a phosphopantetheinyl transferase (PPTase), wherein the recombinant host cell produces a fatty aldehyde, a fatty alcohol, a fatty acid or a fatty ester when cultured in a culture medium under conditions permissive for the production thereof.
48 . The recombinant host cell of claim 47 , wherein the FadR polypeptide is encoded by a fadR gene obtained from Escherichia, Salmonella, Citrobacter, Enterobacter, Klebsiella, Cronobacter, Yersinia, Serratia, Erwinia, Pectobacterium, Photorhabdus, Edwardsiella, Shewanella , or Vibrio.
49 . The recombinant host cell of claim 47 , wherein the FadR polypeptide comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO: 1.
50 . The recombinant host cell of claim 49 , wherein the FadR polypeptide comprises a mutation at amino acid 219 of SEQ ID NO: 1.
51 . The recombinant host cell of claim 50 , wherein the mutation is a substitution of the amino acid 219 of SEQ ID NO: 1 with an asparagine residue.
52 . The recombinant host cell of claim 47 , wherein the overexpression of the FadR polypeptide is caused by a heterologous promoter and/or a ribosome binding site operably linked to a polynucleotide sequence which encodes the FadR polypeptide.
53 . The recombinant host cell of claim 47 , wherein the host cell is further engineered to overexpress
(a) a carboxylic acid reductase,
(b) a thioesterase, and
(c) an alcohol dehydrogenase.
54 . The recombinant host cell of claim 53 , wherein the carboxylic acid reductase is carB (SEQ ID NO: 8); the thioesterase is tesA (SEQ ID NO: 11); and the alcohol dehydrogenase is YjgB (SEQ ID NO: 10) or AlrAadp1 (SEQ ID NO: 9).
55 . The recombinant host cell of claim 47 , wherein the PPTase is EntD from E. coli MG1655 (SEQ ID NO: 12).