Microorganism and method for the production of 1.2-propanediol based on NADPH dependent acetol reductase and improved NADPH supply
The present invention relates to a recombinant microorganism useful for the production of 1,2-propanediol and process for the preparation of 1,2-propanediol. The microorganism of the invention is modified in a way that the 1,2-propanediol production is improved by enhancing NADPH dependent HAR activity.
1. A method for the production of 1,2-propanediol in a fermentative process comprising the steps:
culturing an E. coli microorganism genetically modified for the production of 1,2-propanediol in an appropriate culture medium comprising a carbohydrate as a source of carbon, and
recovering 1,2-propanediol from said culture medium,
wherein said E. Coli microorganism expresses
a Clostridium beijerinckii adh gene coding for a NADPH dependent acetol reductase or
a gldA gene coding for a NADPH dependent glycerol dehydrogenase having at least 90% sequence identity to the sequence set forth in SEQ ID NO:28, wherein the dehydrogenase comprises a glycine, an alanine, or a valine at the amino acid residue corresponding to position 37 of SEQ ID NO: 28.
2. The method of claim 1 , wherein a NADPH availability is increased in the microorganism by at least one of the following genetic modifications:
a pntAB gene operon coding for a nicotinamide nucleotide transhydrogenase is overexpressed, and/or
a pgi gene coding for a phosphoglucose isomerase is attenuated, and/or
a pfkA gene coding for a phosphofructokinase is attenuated, and/or
a zwf gene coding for a glucose-6-phosphate dehydrogenase is overexpressed, and/or
a yjeF gene coding for an ADP-dependent dehydratase is overexpressed, and/or
a gapN gene coding for a NADP-dependent glyceraldehyde-3-phosphate dehydrogenase is overexpressed, and/or
a mutant lpd* gene coding for a NADP-dependent lipoamide dehydrogenase is overexpressed.
3. The method of claim 1 , wherein an endogenous gldA gene coding for an endogenous NADH dependent glycerol dehydrogenase is deleted in the microorganism.
4. The method of claim 1 , wherein the microorganism further comprises a deletion of a yqhD gene coding for a methylglyoxal reductase.
5. The method of claim 1 , wherein said genetically modified microorganism overexpresses the gldA gene coding for the GldA NADPH dependent glycerol dehydrogenase having at least 90% sequence identity to the sequence set forth in SEQ ID NO: 28, wherein the dehydrogenase further comprises a serine or a threonine at the amino acid residue corresponding to position 161 of SEQ ID NO: 28.
6. The method of claim 1 , wherein said genetically modified microorganism overexpresses the gldA gene coding for the GldA NADPH dependent glycerol dehydrogenase having at least 90% sequence identity to the sequence set forth in SEQ ID NO: 28, wherein the dehydrogenase further comprises an alanine, a glycine, or a valine at the amino acid residue corresponding to position 164 of SEQ ID NO: 28.
7. The method of claim 1 , wherein said genetically modified microorganism overexpresses the gldA gene coding for the GldA NADPH dependent glycerol dehydrogenase having at least 90% sequence identity to the sequence set forth in SEQ ID NO: 28, wherein the dehydrogenase further comprises a threonine at the amino acid residue corresponding to position 160 of SEQ ID NO: 28.
8. The method of claim 1 , wherein said genetically modified microorganism overexpresses the gldA gene coding for the GldA NADPH dependent glycerol dehydrogenase having at least 90% sequence identity to the sequence set forth in SEQ ID NO: 28, wherein the dehydrogenase further comprises a serine at the amino acid residue corresponding to position 161 of SEQ ID NO: 28 and an alanine at the amino acid residue corresponding to position 164 of SEQ ID NO: 28.
9. The method of claim 1 , wherein said genetically modified microorganism overexpresses the gldA gene coding for the GldA NADPH dependent glycerol dehydrogenase having at least 90% sequence identity to the sequence set forth in SEQ ID NO: 28, wherein the dehydrogenase further comprises a serine at the amino acid residue corresponding to position 161 of SEQ ID NO: 28, an alanine at the amino acid residue corresponding to position 164 of SEQ ID NO: 28, and a threonine at the amino acid residue corresponding to position 160 of SEQ ID NO: 28.
10. The method of claim 1 , wherein the microorganism further comprises the genes scrK, scrYAB and scrR.
11. The method of claim 1 , wherein the source of carbon is derived from renewable feed-stock.
12. The method of claim 1 , wherein the carbohydrate is selected among the group consisting of glucose, fructose, mannose, xylose, arabinose, galactose, sucrose, cellobiose, maltose, lactose, raffinose, stachyose, maltodextrins, cellulose, hemicellulose, starch, methanol, formaldehyde and glycerol.
13. The method of claim 12 , wherein the carbohydrate is glucose or sucrose.