Metabolically engineered E. coli for enhanced production of oxaloacetate-derived biochemicals
AssigneeUniversity of Georgia Research Foundation, Inc.
InventorsRavi R. Gokarn, Mark A. Eiteman, Elliot Altman
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Abstract
Metabolic engineering is used to increase the carbon flow toward oxaloacetate to enhance production of bulk biochemicals, such as lysine and succinate, in bacterial fermentations. Carbon flow is redirected by genetically engineering the cells to overexpress the enzyme pyruvate carboxylase.
Claims10
A metabolically engineered E. coli cell transformed with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter, wherein said polynucleotide sequence is expressed and produces an enzymatically active pyruvate carboxylase.
A metabolically engineered E. coli cell transformed with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter;
- wherein the cell comprises enzymatically active pyruvate carboxylase and wherein the polynucleotide sequence is chromosomally integrated.
A method for making a metabolically engineered E. coli cell comprising transforming an E. coli cell with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter to yield a metabolically engineered E. coli cell that comprises enzymatically active pyruvate carboxylase.
A method for making succinate comprising:
- (a) providing a first E. coli cell that produces succinate;
- (b) transforming the first E. coli cell with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter to yield a metabolically engineered E. coli cell;
- (c) culturing the metabolically engineered E. coli cell to permit expression of the pyruvate carboxylase to cause increased production of succinate relative to a second E. coli cell that has not been transformed with the polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter;
- and
- (d) isolating the succinate produced by the metabolically engineered E. coli cell.
A method for making succinate comprising:
- (a) providing a first, metabolically engineered E. coli cell that produces succinate, wherein the metabolically engineered E. coli cell has been transformed with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter;
- (b) culturing the metabolically engineered E. coli cell under conditions that permit expression of the pyruvate carboxylase to cause increased production of succinate relative to a second E. coli cell that has not been transformed with the polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter;
- and
- (c) isolating the succinate produced by the metabolically engineered E. coli cell.
A method for making threonine comprising:
- (a) providing a firsts metabolically engineered E. coli cell that produces threonine, wherein the metabolically engineered E. coli cell has been transformed with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter,
- (b) culturing the metabolically engineered E. coli cell under conditions that permit expression of pyruvate carboxylase to cause increased production of threonine relative to a second E. coli cell that has not been transformed with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter;
- and
- (c) isolating the threonine produced by the metabolically engineered E. coli cell.
A method for making threonine comprising:
- (a) providing first E. coli cell that produces threonine;
- (b) transforming the first E. coli cell with a polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter to yield a metabolically engineered E. coli cell;
- (c) culturing the metabolically engineered E. coli cell to permit expression of the pyruvate carboxylase to cause increased production of threonine relative to a second E. coli cell that has not been transformed with the polynucleotide sequence encoding a pyruvate carboxylase operatively linked to a promoter;
- and
- (d) isolating the threonine produced by the metabolically engineered E. coli cell.
Dependencies are inferred from the claim text.