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- Nachgewiesen in: USPTO Patent Applications
- Sprachen: English
- Document Number: 20160333308
- Publication Date: November 17, 2016
- Appl. No: 15/112364
- Application Filed: January 09, 2015
- Assignees: University of Delaware (Newark, DE, US)
- Claim: 1. A non-naturally occurring microbe capable of growing in a medium comprising methanol, wherein the methanol contributes to at least 40% of the carbon source for the non-naturally occurring microbe, expressing heterologous methanol dehydrogenase (MDH) and one or more heterologous ribulose monophosophate (RuMP) pathway enzymes.
- Claim: 2. The non-naturally occurring microbe of claim 1, wherein the expression of the one or more RuMP pathway enzymes is under the control of a formaldehyde responsive promoter.
- Claim: 3. The non-naturally occurring microbe of claim 1, wherein the non-naturally occurring microbe expresses heterologous MDH, heterologous 3-hexulose-6-phosphate synthase (HPS), and heterologous 3-hexulose-6-phosphate isomerase (PHI).
- Claim: 4. The non-naturally occurring microbe of claim 1, wherein the non-naturally occurring microbe contains a deletion of a frmRAB operon.
- Claim: 5. The non-naturally occurring microbe of any one of claims 1-4, further expressing one or more heterologous pentose-phosphate pathway (PPP) enzymes.
- Claim: 6. The non-naturally occurring microbe of claim 5, wherein the expression of the one or more heterologous PPP enzymes is under the control of a formaldehyde responsive promoter.
- Claim: 7. The non-naturally occurring microbe of claim 5, wherein the non-naturally occurring microbe expresses heterologous phosphofructokinase (PFK), heterologous fructose bisphosphate aldolase (FBA), heterologous transketolase (TKT), heterologous fructose/sedoheptulose biphosphatase (GLPX), heterologous transaldolase (TAL), heterologous ribose-5-phospate (RPI) and heterologous ribulose phosphate epimerase (RPE).
- Claim: 8. The non-naturally occurring microbe of any one of claims 1-7, further expressing one or more heterologous cyclic formaldehyde dissimilation enzymes.
- Claim: 9. The non-naturally occurring microbe of claim of claim 8, wherein the expression of the one or more heterologous cyclic formaldehyde dissimilation enzymes is under the control of a formaldehyde responsive promoter.
- Claim: 10. The non-naturally occurring microbe of claim of claim 8, wherein the non-naturally occurring microbe expresses heterologous glucose-6-phosphate isomerase (PGI), glucose-6-phosphate-1-dehydrogenase (ZWF), 6-phosphogluconolactonase (PGL), and 6-phosphogluconate dehydrogenase (GND).
- Claim: 11. The non-naturally occurring microbe of claim 8, wherein the non-naturally occurring microbe contains a deletion of a phosphogluconate dehydratase gene (edd).
- Claim: 12. The non-naturally occurring microbe of any one of claims 1-11, further expressing one or more heterologous CO2 fixation pathway enzymes.
- Claim: 13. The non-naturally occurring microbe of claim 12, wherein the expression of the one or more heterologous CO2 fixation pathway enzymes is under the control of a formaldehyde responsive promoter.
- Claim: 14. The non-naturally occurring microbe of claim 12, wherein the non-naturally occurring microbe expresses heterologous carbonic anhydrase (CA), heterologous formate dehydrogenase (FDH), and heterologous formaldehyde dehydrogenase (FLD).
- Claim: 15. The non-naturally occurring microbe of any one of claims 1-14, further expressing heterologous dihydroxyacetone synthase (DHAS) and heterologous dihydroxyacetone kinase (DAK).
- Claim: 16. The non-naturally occurring microbe of any one of claims 1-15, wherein the non-naturally occurring microbe is derived from a microbe selected from the group consisting of facultative aerobic organisms, facultative anaerobic organisms, and anaerobic organisms.
- Claim: 17. The non-naturally occurring microbe of any one of claims 1-15, wherein the non-naturally occurring microbe is derived from a microbe selected from the group consisting of phyla Proteobacteria, Firmicutes, Actinobacteria, Cyanobacteria, Chlorobi, and Deinococcus-Thermus.
- Claim: 18. The non-naturally occurring microbe of any one of claims 1-15, wherein the non-naturally occurring microbe is derived from a microbe selected from the group consisting of Escherichia, Bacillus, Clostridium, Enterobacter, Klebsiella, Enterobacteria, Mannheimia, Pseudomonas, Acinetobacter, Shewanella, Ralstonia, Geobacter, Zymomonas, Acetobacter, Geobacillus, Lactococcus, Streptococcus, Lactobacillus, Corynebacterium, Streptomyces, Propionibacterium, Synechocystis, Synechococcus, Cyanobacteria, Chlorobi, and Deinococcus.
- Claim: 19. The non-naturally occurring microbe of any one of claims 1-15, wherein the non-naturally occurring microbe is E. coli.
- Claim: 20. A method for producing a metabolite, comprising growing a non-naturally occurring microbe of any one of claims 1-19 in a medium comprising methanol, wherein the methanol contributes to at least 40% of the carbon source for the non-naturally occurring microbe, whereby the metabolite is produced.
- Claim: 21. The method of claim 20, wherein the metabolite is selected from the group consisting of 4-carbon chemicals, diacids, 3-carbon chemicals, higher carboxylic acids, alcohols of higher carboxylic acids, and polyhydroxyalkanoates.
- Claim: 22. The method of claim 20, wherein the metabolite is n-butanol.
- Claim: 23. The method of any one of claims 20-22, further comprising modifying the heterologous MDH or any one of the one or more heterologous RuMP pathway enzymes, whereby the production of the metabolite is improved.
- Claim: 24. The method of any one of claims 20-23, further comprising fixing CO2.
- Claim: 25. The method of any one of claims 20-24, wherein the non-naturally occurring microbe is grown at a temperature higher than 37° C.
- Claim: 26. The method of any one of claims 20-25, wherein the non-naturally occurring microbe is grown anaerobically.
- Claim: 27. The method of any one of claims 20-26, whereby at least 40% of the carbon in the metabolite is derived from the methanol.
- Claim: 28. The method of any one of claims 20-27, wherein the metabolite is an amino acid or tricarboxylic acid (TCA) intermediate having a carbon at the fourth position derived from the methanol.
- Claim: 29. A method for making a non-naturally occurring microbe capable of growing in a medium comprising methanol, wherein the methanol contributes to at least 40% of the carbon source for the non-naturally occurring microbe, comprising expressing heterologous methanol dehydrogenase (MDH) and one or more heterologous ribulose monophosophate (RuMP) pathway enzymes in a non-methylotrophic microbe, whereby the non-naturally occurring microbe is obtained.
- Claim: 30. The method of claim 29, wherein the non-naturally occurring microbe expresses the heterologous MDH, heterologous 3-hexulose-6-phosphate synthase (HPS), and heterologous 3-hexulose-6-phosphate isomerase (PHI).
- Claim: 31. The method of claim 29 or 30, further comprising expressing one or more heterologous pentose-phosphate pathway (PPP) enzymes in the non-methylotrophic microbe.
- Claim: 32. The method of claim 31, wherein the non-methylotrophic microbe expresses heterologous phosphofructokinase (PFK), heterologous fructose bisphosphate aldolase (FBA), heterologous transketolase (TKT), transaldolase (TAL), heterologous fructose/sedoheptulose biphosphatase (GLPX), heterologous ribose-5-phosphate (RPI), and heterologous ribulose phosphate epimerase (RPE).
- Claim: 33. The method of any one of claims 29-32, further comprising expressing one or more heterologous cyclic formaldehyde dissimilation enzymes.
- Claim: 34. The method of claim 33, wherein the non-naturally occurring microbe expresses heterologous glucose-6-phosphate isomerase (PGI), glucose-6-phosphate-1-dehydrogenase (ZWF), 6-phosphogluconolactonase (PGL), and 6-phosphogluconate dehydrogenase (GND).
- Claim: 35. The method of any one of claims 29-34, further comprising expressing one or more heterologous CO2 fixation pathway enzymes in the non-methylotrophic microbe.
- Claim: 36. The method of claim 35, wherein the non-methylotrophic microbe expresses heterologous carbonic anhydrase (CA), heterologous formate dehydrogenase (FDH), and heterologous formaldehyde dehydrogenase (FLD).
- Claim: 37. The method of any one of claims 29-36, further comprising expressing heterologous dihydroxyacetone synthase (DHAS) and dihydroxyacetone kinase (DAK).
- Current International Class: 12; 12; 12; 12; 12
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