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.
Claims34
A genetically modified E. coli microorganism comprising:
- a production pathway comprising citramalate synthase for the production of citramalate,
- a conditionally triggered synthetic metabolic valve that silences gene expression of the citrate synthase (gltA), SoxS, and/or glucose-6-phosphate-dehydrogenase (zwf) gene(s);
- or
- a conditionally triggered synthetic metabolic valve that enables selective proteolysis of the citrate synthase (gltA), SoxS, and/or glucose-6-phosphate-dehydrogenase (zwf) enzyme(s);
- wherein the synthetic metabolic valve(s) of 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 increased within the genetically modified microorganism under aerobic or partially aerobic conditions during the stationary phase or non-dividing cellular state to produce an acetyl CoA pool;
- and
- wherein sugar uptake is enhanced within the genetically modified microorganism, when compared to a non-genetically modified microorganism.
The genetically modified microorganism of claim 4, wherein the pyruvate-flavodoxin/ferredoxin oxidoreductase enzyme is encoded by the ydbK.
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
A genetically modified microorganism comprising:
- a production pathway comprising at least one enzyme for producing a product from an acetyl CoA precursor, and
- a conditionally triggered synthetic metabolic valve that silences gene expression of the citrate synthase (gltA), SoxS, and/or glucose-6-phosphate-dehydrogenase (zwf) gene(s);
- and
- a conditionally triggered synthetic metabolic valve that enables selective proteolysis of the citrate synthase (gltA), SoxS, and/or glucose-6-phosphate-dehydrogenase (zwf) enzyme(s);
- and
- deletion of endogenous poxB and pjlB genes;
- wherein the synthetic metabolic valve(s) of the microorganism are conditionally triggered during the 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, 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 increased within the genetically modified microorganism under aerobic or partially aerobic conditions during the stationary phase or non-dividing cellular state to produce an acetyl CoA pool;
- and
- wherein sugar uptake is enhanced within the genetically modified microorganism, when compared to a non-genetically modified microorganism.
(canceled)
-26. (canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
(canceled)
Dependencies are inferred from the claim text.