Microorganisms and methods for the production of oxygenated compounds from hexoses

Patent metadata

Field	Value
Publication number	US-12006526-B2
Application number	US-202016796417-A
Country	US
Kind code	B2
Filing date	Feb 20, 2020
Priority date	Feb 20, 2019
Publication date	Jun 11, 2024
Grant date	Jun 11, 2024

Abstract

The present application relates to recombinant microorganisms useful in the biosynthesis of monoethylene glycol (MEG), or optionally MEG and one or more co-product, from one or more hexose feedstock. The present application also relates to recombinant microorganisms useful in the biosynthesis of glycolic acid (GA), or optionally GA and one or more co-product, from one or more hexose feedstock. The present application relates to recombinant microorganisms useful in the biosynthesis of xylitol, or optionally xylitol and one or more co-product, from one or more hexose feedstock. Also provided are methods of producing MEG (or GA or xylitol), or optionally MEG (or GA or xylitol) and one or more co-product, from one or more hexose feedstock using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or the products MEG (or GA or xylitol), or optionally MEG (or GA or xylitol) and one or more co-product.

First claim

What is claimed is:

1. A recombinant microorganism that produces glycolic acid (GA) from one or more hexose feedstocks via
D-xylulose-5-phosphate and/or D-ribulose-5-phosphate;
wherein the recombinant microorganism comprises one or more overexpressed enzymes for the conversion of D-xylulose-5-phosphate and/or D-ribulose-5-phosphate to D-xylulose-1-phosphate or D-ribulose-1-phosphate,
wherein the one or more overexpressed enzymes is one or more enzymes having phosphopentomutase activity;
wherein the recombinant microorganism further comprises at least one overexpressed enzyme having transketolase activity or at least one overexpressed enzyme having fructose-6-phosphate phosphoketolase activity;
wherein the recombinant microorganism further comprises a deleted or diminished activity in one or more endogenous enzymes selected from glucose 6-phosphate-1-dehydrogenase (zwf), 6-phosphogluconolactonase (pgl), or 6-phosphogluconate dehydrogenase (gnd); and
wherein the recombinant microorganism is a recombinant E. coli.

2. The recombinant microorganism of claim 1 , wherein one or more co-products is co-produced with GA, wherein the one or more co-products is selected from the group consisting of acetone, isopropanol, propene, and isobutene.

3. The recombinant microorganism of claim 1 , wherein the recombinant microorganism comprises at least one overexpressed enzyme having transketolase activity, wherein the at least one overexpressed enzyme having transketolase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 148 or SEQ ID NO: 150.

4. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises at least one overexpressed enzyme having transaldolase activity, wherein the at least one overexpressed enzyme having transaldolase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 152 or SEQ ID NO: 154.

5. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises at least one overexpressed enzyme having ribulose-5-phosphate 3-epimerase activity, wherein the at least one overexpressed enzyme having ribulose-5-phosphate 3-epimerase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 158.

6. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises at least one overexpressed enzyme having ribose-5-phosphate isomerase activity, wherein the at least one overexpressed enzyme having ribose-5-phosphate isomerase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 156 or SEQ ID NO: 253.

7. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises a deleted or diminished activity in at least one endogenous enzyme selected from glyceraldehyde 3-phosphate dehydrogenase (gapA), phosphoglycerate kinase (pgk) and/or phosphoglycerate mutase (gpmA/gpmM).

8. The recombinant microorganism of claim 1 , wherein the recombinant microorganism comprises at least one overexpressed enzyme having fructose-6-phosphate phosphoketolase activity, wherein the at least one overexpressed enzyme having fructose-6-phosphate phosphoketolase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 212, SEQ ID NO: 214, SEQ ID NO: 216, or SEQ ID NO: 218.

9. The recombinant microorganism of claim 8 , wherein the recombinant microorganism further comprises at least one overexpressed enzyme having phosphate acetyltransferase activity, wherein the at least one enzyme having phosphate acetyltransferase activity comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 220 or SEQ ID NO: 222.

10. The recombinant microorganism of claim 1 , wherein the recombinant microorganism further comprises a deleted or diminished activity in an endogenous 6-phosphofructokinase enzyme (pfkA/pfkB).

11. The recombinant microorganism of claim 1 , wherein the one or more enzymes having phosphopentomutase activity is Pgm3.

12. The recombinant microorganism of claim 1 , wherein the overexpressed enzyme having transketolase activity or the overexpressed enzyme having fructose-6-phosphate phosphoketolase activity enables a conversion of one or more hexose feedstocks to D-xylulose-5-phosphate and/or D-ribulose-5-phosphate without loss of carbon.

13. The recombinant microorganism of claim 1 , wherein GA is produced through the conversion of glycolaldehyde in a C2 pathway and through the conversion of dihydroxyacetone phosphate (DHAP) or pyruvate in a C3 pathway.

14. The recombinant microorganism of claim 1 , wherein GA is produced through the conversion of glycolaldehyde in a C2 pathway and one or more co-products is produced through the conversion of dihydroxyacetone phosphate (DHAP) or pyruvate in a C3 pathway.

15. A method of producing glycolic acid (GA) using a recombinant microorganism of claim 1 , wherein the method comprises cultivating the recombinant microorganism in a culture medium containing one or more hexose feedstocks providing a carbon source until the GA is produced.

16. A method of producing a recombinant microorganism that produces or accumulates glycolic acid (GA) from one or more exogenous hexose feedstocks via one or more pentose-5-phosphate intermediates, comprising:
expressing in the recombinant microorganism one or more enzymes for the conversion of the one or more hexose feedstocks to the one or more pentose-5-phosphate intermediates, wherein the one or more pentose-5-phosphate intermediates is D-xylulose-5-phosphate and/or D-ribulose-5-phosphate;
overexpressing in the recombinant microorganism one or more enzymes for the conversion of D-xylulose-5-phosphate and/or D-ribulose-5-phosphate to D-xylulose-1-phosphate or D-ribulose-1-phosphate, wherein the one or more overexpressed enzymes is one or more enzyme having phosphopentomutase activity;
overexpressing in the recombinant microorganism at least one enzyme having transketolase activity or at least one enzyme having fructose-6-phosphate phosphoketolase activity;
deleting or diminishing activity in the recombinant microorganism of one or more endogenous enzymes selected from glucose 6-phosphate-1-dehydrogenase (zwf), 6-phosphogluconolactonase (pgl), or 6-phosphogluconate dehydrogenase (gnd);
expressing in the recombinant microorganism a C2 pathway comprising one or more enzymes for the production of GA from glycolaldehyde; and
expressing in the recombinant microorganism a C3 pathway comprising one or more enzymes for the production of GA derived from DHAP or pyruvate; and
culturing the recombinant microorganism in a culture medium containing the one or more hexose feedstocks to produce or accumulate GA,
wherein glycolaldehyde and DHAP (or pyruvate) are intermediates produced in the C2 pathway, and wherein GA is produced in both the C2 and C3 pathways.

17. A method of producing a recombinant microorganism that produces or accumulates glycolic acid (GA) and one or more co-products from one or more exogenous hexose feedstocks via one or more pentose-5-phosphate intermediates, comprising:
expressing in the recombinant microorganism one or more enzymes for the conversion of the one or more hexose feedstocks to the one or more pentose-5-phosphate intermediates, wherein the one or more pentose-5-phosphate intermediates is D-xylulose-5-phosphate and/or D-ribulose-5-phosphate;
overexpressing in the recombinant microorganism one or more enzymes for the conversion of D-xylulose-5-phosphate and/or D-ri

Assignees

• Braskem Sa

Inventors

• Koch Daniel Johannes
• Parizzi Lucas Pedersen
• Galzerani Felipe

Classifications

• C12R2001/19

  Escherichia coli · CPC title

• C12N1/205

  Bacterial isolates · CPC title

• C12Y504/02

  Phosphotransferases (phosphomutases) (5.4.2) · CPC title

• C12Y503/01006

  Ribose-5-phosphate isomerase (5.3.1.6) · CPC title

• C12Y501/03001

  Ribulose-phosphate 3-epimerase (5.1.3.1) · CPC title