Production of propanols, alcohols, and polyols in consolidated bioprocessing organisms

Patent metadata

Field	Value
Publication number	US-11634735-B2
Application number	US-202016918415-A
Country	US
Kind code	B2
Filing date	Jul 1, 2020
Priority date	Aug 21, 2009
Publication date	Apr 25, 2023
Grant date	Apr 25, 2023

Abstract

The present invention provides for novel metabolic pathways leading to propanol, alcohol or polyol formation in a consolidated bioprocessing system (CBP), where lignocellulosic biomass is efficiently converted to such products. More specifically, the invention provides for a recombinant microorganism, where the microorganism expresses one or more native and/or heterologous enzymes; where the one or more enzymes function in one or more engineered metabolic pathways to achieve: (1) conversion of a carbohydrate source to 1,2-propanediol, isopropropanol, ethanol and/or glycerol; (2) conversion of a carbohydrate source to n-propanol and isopropanol; (3) conversion of a carbohydrate source to isopropanol and methanol; or (4) conversion of a carbohydrate source to propanediol and acetone; wherein the one or more native and/or heterologous enzymes is activated, upregulated or downregulated.

First claim

The invention claimed is:

1. A recombinant microorganism;
comprising a plurality of heterologous enzymes, wherein said plurality of heterologous enzymes are expressed;
wherein said plurality of heterologous enzymes function in one or more engineered metabolic pathways to convert a carbohydrate source to n-propanol, isopropanol and, optionally, ethanol; and
wherein said plurality of heterologous enzymes is activated or upregulated, and comprises:
a heterologous pyruvate formate lyase,
a heterologous methylglyoxal synthase, and
a heterologous vitamin B12-independent diol dehydratase;
wherein said recombinant microorganism is a yeast.

2. The recombinant microorganism of claim 1 , wherein said carbohydrate source is lignocellulose.

3. The recombinant microorganism of claim 1 , wherein said carbohydrate source is converted to dihydroxyacetone phosphate and glyceraldehyde phosphate.

4. The recombinant microorganism of claim 1 , wherein one of said engineered metabolic pathways comprises conversion of pyruvate to isopropanol.

5. The recombinant microorganism of claim 1 , wherein one of said engineered metabolic pathway comprises conversion of pyruvate to ethanol.

6. The recombinant microorganism of claim 1 , wherein the engineered metabolic pathway that produces isopropanol comprises: (a) conversion of acetyl-CoA to acetate;
(b) conversion of acetyl-CoA to acetoacetyl-CoA; (c) conversion of acetoacetyl-CoA to acetoacetate; (d) conversion of acetoacetate to acetone; and (e) reduction of acetone to isopropanol.

7. The recombinant microorganism of claim 1 , wherein the engineered metabolic pathway that produces ethanol comprises: (a) conversion of acetyl-CoA to ethanol.

8. The recombinant microorganism of claim 1 , wherein said recombinant microorganism is selected from the group consisting of Saccharomyces cerevisiae, Kluyveromyces lactis, Kluyveromyces marxianus, Pichia pastoris, Yarrowia lipolytica , Hansemila polymorphs, Phaffia rhodozyma, Candida ittliis, Arxula adeninivorans, Pichia stipitis, Debaryomyces hansenii, Debaryomyces polymorphic, Schizosaccharomyces pombe, Candida albicans , and Schwanniomyces occidentalis.

9. The recombinant microorganism of claim 3 , wherein one of said engineered metabolic pathways comprises the conversion of dihydroxyacetone phosphate to n-propanol.

10. The recombinant microorganism of claim 9 , wherein said dihydroxyacetone phosphate is converted to methylglyoxal with the heterologous methylglyoxal synthase.

11. The recombinant microorganism of claim 10 , wherein said methylglyoxal is converted to acetol by an aldo-keto reductase or converted to lactaldehyde by a glyoxylate reductase, the heterologous methylglyoxal dehydrogenase or an aldehyde dehydrogenase.

12. The recombinant microorganism of claim 11 , wherein said acetol is converted to propanediol by an aldo-keto reductase or said lactaldehyde is converted to propanediol by an aldehyde reductase.

13. The recombinant microorganism of claim 12 , wherein said propanediol is converted to propanal with the heterologous vitamin B12-independent diol dehydratase.

14. The recombinant microorganism of claim 13 , wherein said propanal is reduced to n-propanol by a dehydrogenase (E.C.1.1.1.202).

15. The recombinant microorganism of claim 6 , wherein acetyl-CoA is converted to acetoacetyl-CoA by a thiolase (E.C.2.3.1.9).

16. The recombinant microorganism of claim 15 , wherein said acetoacetyl-CoA is converted to acetoacetate by a CoA transferase.

17. The recombinant microorganism of claim 16 , wherein said acetoacetate is converted to acetone by an acetoacetate decarboxylase.

18. The recombinant microorganism of claim 17 , wherein said acetone is reduced to isopropanol by an isopropanol dehydrogenase or an alcohol dehydrogenase.

19. The recombinant microorganism of claim 1 , wherein the vitamin B12-independent diol dehydratase is from Clostridium sp., Roseburia sp. or Klebsiella sp.

20. The recombinant microorganism of claim 13 , wherein the vitamin B12-independent diol dehydratase is from Clostridium butyricum, Clostridium glycolicum or Roseburia inulivorans.

Assignees

• Lallemand Hungary Liquidity Man Llc

Inventors

• Mcbride John E
• Rajgarhia Vineet
• Shaw Iv Arthur J
• Tripathi Shital A
• Brevnova Elena
• Caiazza Nicky
• Van Dijken Johannes Pieter
• Froehlich Allan C
• Sillers William Ryan
• Flatt James H

Classifications

• C12P7/18 Primary

  polyhydric · CPC title

• C12P2203/00

  Fermentation products obtained from optionally pretreated or hydrolyzed cellulosic or lignocellulosic material as the carbon source · CPC title

• C12P7/04

  acyclic · CPC title

• C12P7/36

  produced from substrate containing grain or cereal material · CPC title

• Y02E50/10

  Biofuels, e.g. bio-diesel · CPC title