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US 2021/0324431 A1Published application
Production Of Odd Chain Fatty Acid Derivatives In Recombinant Microbial Cells
AssigneeGenomatica, Inc.
InventorsGrace J. LEE, John HALIBURTON, Zhihao HU, Andreas W. SCHIRMER, Kevin HOLDEN
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Abstract
Recombinant microbial cells are provided which have been engineered to produce fatty acid derivatives having linear chains containing an odd number of carbon atoms by the fatty acid biosynthetic pathway. Also provided are methods of making odd chain fatty acid derivatives using the recombinant microbial cells, and compositions comprising odd chain fatty acid derivatives produced by such methods.
Claims20
A recombinant microbial cell comprising:
- (a) polynucleotides encoding polypeptides having enzymatic activity effective to produce an increased amount of propionyl-CoA in the recombinant microbial cell relative to the amount of propionyl-CoA produced in a parental microbial cell lacking or having a reduced amount of said enzymatic activity, wherein the polynucleotides encode
- (i) a polypeptide having (R)-citramalate synthase activity, a polypeptide having isopropylmalate isomerase activity, and a polypeptide having beta-isopropylmalate dehydrogenase activity, and/or
- (ii) a polypeptide having aspartokinase activity, a polypeptide having homoserine dehydrogenase activity, a polypeptide having homoserine kinase activity, a polypeptide having threonine synthase activity, and a polypeptide having threonine deaminase activity,
- (b) a polynucleotide encoding a polypeptide having β-ketoacyl-ACP synthase activity that utilizes propionyl-CoA as a substrate and has 80% sequence identity to SEQ ID NO:2, and
- (c) a polynucleotide encoding a polypeptide having fatty acid derivative enzyme activity,
- wherein the recombinant microbial cell produces a fatty acid derivative composition comprising odd chain and even chain fatty acid derivatives when cultured in the presence of a carbon source under conditions effective to express the polynucleotides according to (a), (b), and (c), and
- wherein at least 10% of the fatty acid derivatives in the fatty acid derivative composition are odd chain fatty acid derivatives.
4 more elements
(canceled)
(canceled)
A method of making a recombinant microbial cell which produces a higher titer or higher proportion of odd chain fatty acid derivatives than produced by a parental microbial cell, the method comprising:
- obtaining a parental microbial cell comprising a polynucleotide encoding a polypeptide having β-ketoacyl-ACP synthase activity that utilizes propionyl-CoA as a substrate and a polynucleotide encoding a polypeptide having fatty acid derivative enzyme activity, and
- engineering the parental microbial cell to obtain a recombinant microbial cell which produces or is capable of producing a greater amount of propionyl-CoA than the amount of propionyl-CoA produced by the parental microbial cell when cultured under the same conditions, wherein the step of engineering the parental microbial cell comprises:
- engineering the parental microbial cell to express polynucleotides encoding:
- (a) polypeptides:
- (i) having aspartokinase activity, homoserine dehydrogenase activity, homoserine kinase activity, threonine synthase activity, and threonine deaminase activity;
- and/or
- (ii) polypeptides having (R)-citramalate synthase activity, isopropylmalate isomerase activity, and beta-isopropylmalate dehydrogenase activity;
- and
- (b) a polypeptide having β-ketoacyl-ACP synthase activity that utilizes propionyl-CoA as a substrate and has 80% sequence identity to SEQ ID NO:2;
- and
- (c) a polypeptide having fatty acid derivative enzyme activity;
- wherein the recombinant microbial cell produces a higher titer or higher proportion of odd chain fatty acid derivatives when cultured in the presence of a carbon source under conditions effective to express the polynucleotides, relative to the titer or proportion of odd chain fatty acid derivatives produced by the parental microbial cell cultured under the same conditions.
9 more elements
(canceled)
A method of increasing the titer or the proportion of odd chain fatty acid derivatives produced by a microbial cell, the method comprising:
- obtaining a parental microbial cell which produces fatty acid derivatives, and
- engineering the parental microbial cell to obtain a recombinant microbial cell which produces or is capable of producing a greater amount of propionyl-CoA than the amount of propionyl-CoA produced by the parental microbial cell when cultured under the same conditions, wherein the step of engineering the parental microbial cell comprises:
- engineering the parental microbial cell to express polynucleotides encoding:
- (a) polypeptides:
- (i) having aspartokinase activity, homoserine dehydrogenase activity, homoserine kinase activity, threonine synthase activity, and threonine deaminase activity;
- and/or
- (ii) polypeptides having (R)-citramalate synthase activity, isopropylmalate isomerase activity, and beta-isopropylmalate dehydrogenase activity;
- and
- (b) a polypeptide having β-ketoacyl-ACP synthase activity that utilizes propionyl-CoA as a substrate and has 80% sequence identity to SEQ ID NO:2;
- and
- (c) a polypeptide having fatty acid derivative enzyme activity;
- wherein the recombinant microbial cell produces a higher titer or higher proportion of odd chain fatty acid derivatives when cultured in the presence of a carbon source under conditions effective to produce propionyl-CoA and fatty acid derivatives in the recombinant microbial cell, relative to the titer or proportion of odd chain fatty acid derivatives produced by the parental microbial cell cultured under the same conditions.
9 more elements
Dependencies are inferred from the claim text.