Back to results
US 12,760,336 B2Granted
Methods and compositions for the production of acetyl-CoA derived products
AssigneeDuke University
InventorsMichael David Lynch, Shuai Li
Life of this patent
Estimating the term…
Abstract
The present disclosure is related to genetically engineered microbial strains and related bioprocesses for the production of products from acetyl-CoA. Specifically, the use of dynamically controlled synthetic metabolic valves to reduce the activity of certain enzymes, leads to increased product production in a two-stage process.
Claims17
A genetically modified E. coli microorganism comprising:
- a production pathway comprising citramalate synthase for the production of citramalate,
- a conditional induction of SoxS by depletion of a limiting nutrient from a growth medium in which the genetically modified microorganism is growing;
- and
- a conditionally triggered synthetic metabolic valve that silences gene expression of the glucose-6-phosphate-dehydrogenase (zwf) gene;
- or
- a conditionally triggered synthetic metabolic valve that enables selective proteolysis of the glucose-6-phosphate-dehydrogenase (zwf) enzyme;
- wherein the synthetic metabolic valve(s) and induction of SoxS in the microorganism are conditionally triggered during a stationary phase or non-dividing cellular state;
- wherein, under conditions of depleting of a limiting nutrient from a growth medium in which the genetically modified microorganism is growing, a stationary phase or non-dividing cellular state is induced;
- wherein pyruvate-flavodoxin/ferredoxin oxidoreductase enzyme activity is overexpressed upon depletion of a limiting nutrient from a growth medium in which the genetically modified microorganism is growing during the stationary phase or non-dividing cellular state to produce an acetyl CoA pool;
- and
- wherein PTS-dependent glucose uptake is enhanced within the genetically modified microorganism, when compared to a non-genetically modified microorganism.
8 more elements
A genetically modified E. coli microorganism comprising:
- a production pathway comprising at least one enzyme for producing a product from an acetyl CoA precursor, and
- a conditional induction of SoxS depletion of a limiting nutrient from a growth medium in which the genetically modified microorganism is growing;
- a conditionally triggered synthetic metabolic valve that silences gene expression of the glucose-6-phosphate-dehydrogenase (zwf) gene;
- or
- a conditionally triggered synthetic metabolic valve that enables selective proteolysis of the glucose-6-phosphate-dehydrogenase (zwf) enzyme;
- deletion of endogenous pyruvate oxidase (poxB) and pyruvate-formate lyase (pflB) genes genes;
- wherein the synthetic metabolic valve(s) and induction of SoxS in the microorganism are conditionally triggered during a stationary phase or non-dividing cellular state;
- wherein, under conditions of depleting of a limiting nutrient from a growth medium in which the genetically modified microorganism is growing, a stationary phase or non-dividing cellular state is induced;
- wherein pyruvate-flavodoxin/ferredoxin oxidoreductase enzyme activity is overexpressed upon depletion of a limiting nutrient from a growth medium in which the genetically modified microorganism is growing during the stationary phase or non-dividing cellular state to produce an acetyl CoA pool;
- and
- wherein PTS-dependent glucose uptake is enhanced within the genetically modified microorganism, when compared to a non-genetically modified microorganism.
8 more elements
Dependencies are inferred from the claim text.