Metabolic pathways with increased carbon yield
The present invention relates to the conversion of a carbon source into acetyl phosphate with increased carbon yield. In particular, the invention provides metabolically engineered micro-organisms capable of producing acetyl phosphate from a carbon source with increased carbon yield, which micro-organisms have been transformed with at least one exogenous nucleic acid encoding a phosphoketolase having sedoheptulose-7-phosphate phosphoketolase activity and which are further genetically modified to have eliminated transketolase activity. The invention also provides methods for the production of chemicals using said micro-organisms.
1. A metabolically engineered micro-organism capable of increased acetyl phosphate conversion from a carbon source compared to a corresponding non-metabolically engineered micro-organism, wherein the micro-organism comprises at least one exogenous nucleic acid encoding a phosphoketolase having sedoheptulose-7-phosphate phosphoketolase activity, wherein the phosphoketolase having sedoheptulose-7-phosphate phosphoketolase activity is a Bifidobacterium longum phosphoketolase comprising the sequence set forth in SEQ ID NO: 2, and wherein the micro-organism is further genetically modified to have eliminated transketolase activity.
2. The micro-organism according to claim 1 , wherein the micro-organism is genetically modified to have eliminated transaldolase activity.
3. The micro-organism according to claim 2 , wherein the micro-organism is genetically modified to express or overexpress at least one enzyme selected from the group consisting of an enzyme having fructose-6-phosphoketolase and/or xylulose-5-phosphoketolase activity, a sedoheptulose-1,7-bisphosphate aldolase, a sedoheptulose-1,7-bisphosphatase, a ribose-5-phosphate isomerase, a ribulose-5-phosphate epimerase and a triose phosphate isomerase.
4. The micro-organism according to claim 3 , wherein the micro-organism is genetically modified to express or overexpress a sedoheptulose-1,7-bisphosphatase.
5. The micro-organism according to claim 1 , wherein the micro-organism is genetically modified to express or overexpress at least one enzyme selected from the group consisting of an enzyme having fructose-6-phosphoketolase and/or xylulose-5-phosphoketolase activity, a transaldolase, a ribose-5-phosphate isomerase, a ribulose-5-phosphate epimerase, a triose phosphate isomerase, a fructose 1,6-bisphosphate aldolase, and a fructose 1,6-bisphosphatase.
6. The micro-organism according to claim 5 , wherein the micro-organism is genetically modified to express or overexpress a fructose-1,6-bisphosphatase.
7. The micro-organism according to claim 1 , wherein the micro-organism is genetically modified to have reduced or eliminated activity of one or more of a phosphoglycerate kinase, a phosphofructokinase and a glucose-6-phosphate dehydrogenase.
8. The micro-organism according to claim 1 , wherein the micro-organism is genetically modified to have reduced or eliminated activity of an alkaline phosphatase.
9. The micro-organism according to claim 1 , wherein the micro-organism is further genetically modified to express or overexpress at least one enzyme of a pathway that converts acetyl-phosphate into a molecule of interest, said molecule of interest being selected from the group consisting of polyketides, polyhydroxyalcanoates, hydroxyacids, fatty alcohols, fatty acids, amino acids, acetone, isopropanol, butanol, isobutanol, isobutene, and propene.
10. The micro-organism according to claim 1 , wherein the micro-organism is a bacterium or a yeast.
11. The micro-organism according to claim 10 , wherein the microorganism is a yeast.
12. The micro-organism according to claim 1 , wherein the micro-organism is E. coli or S. cerevisae.
13. The micro-organism according to claim 12 , wherein the microorganism is S. cerevisae.
14. The micro-organism according to claim 1 , wherein the phosphoketolase is encoded by a nucleic acid comprising the sequence set forth in SEQ ID NO: 3.
15. A method for the production of a molecule of interest, said molecule of interest being derived from acetyl-CoA and selected from the group consisting of polyketides, polyhydroxyalcanoates, hydroxyacids, fatty alcohols, fatty acids, amino acids, acetone, isopropanol, butanol, isobutanol, isobutene, and propene, wherein said method comprises:
culturing a metabolically engineered micro-organism according to claim 1 in the presence of a suitable carbon source that can be metabolized by said micro-organism.