Methods and compositions for producing solvents

Patent metadata

Field	Value
Publication number	US-10119115-B2
Application number	US-201514796253-A
Country	US
Kind code	B2
Filing date	Jul 10, 2015
Priority date	May 17, 2007
Publication date	Nov 6, 2018
Grant date	Nov 6, 2018

Abstract

Described herein are methods, compositions and synthetic biology approaches for solvent production, including but not limited to butanol production. Described herein are recombinant bacteria and yeast strains which may be used in production of a solvent, including but not limited to butanol, from lignocellulosic and other plant-based feedstocks. Described herein are methods of producing solvents, including but not limited to butanol, using bacteria and yeast strains. Described herein are methods of producing organisms that display highly efficient butanol production.

First claim

We claim:

1. A recombinant solventogenic microorganism, wherein the microorganism is transformed with:
(a) a nucleic acid molecule comprising at least one Clostridium beijerinckii Adh (alcohol dehydrogenase) polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 14, wherein the polynucleotide encodes a polypeptide having alcohol dehydrogenase activity;
(b) a nucleic acid molecule comprising at least one Clostridium beijerinckii Bcd (butyryl-CoA dehydrogenase) polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 2, wherein the polynucleotide encodes a polypeptide having butyryl-CoA dehydrogenase activity; and
(c) a nucleic acid molecule comprising at least one Clostridium beijerinckii Buk (butyrate kinase) polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 3, wherein the polynucleotide encodes a polypeptide having butyrate kinase activity;
wherein the recombinant solventogenic microorganism is capable of (i) more efficient butanol production, (ii) faster butanol production, and/or (iii) increased butanol production relative to the same recombinant solventogenic microorganism that is not transformed with the nucleic acid molecules of (a), (b), and (c).

2. The recombinant solventogenic microorganism of claim 1 , wherein the microorganism is further transformed with a polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 4, 5, 6 or combinations thereof, and wherein the polynucleotide encodes a polypeptide having chemotaxis protein activity.

3. The recombinant solventogenic microorganism of claim 1 , wherein expression of the Clostridium beijerinckii Adh polynucleotide is controlled by an inducible or a constitutive promoter.

4. The recombinant solventogenic microorganism of claim 1 , which is a yeast, bacterium, or fungi.

5. The recombinant solventogenic microorganism of claim 4 , wherein the bacterium is an Escherichia or a Clostridium.

6. The recombinant solventogenic microorganism of claim 4 , wherein the bacterium is a Clostridium beijerinckii or Clostridium acetobutylicum.

7. The recombinant solventogenic microorganism of claim 4 , wherein the bacterium is a recombinant Clostridium beijerinckii 8052.

8. The recombinant solventogenic microorganism of claim 1 , which is cellulolytic.

9. The recombinant solventogenic microorganism of claim 1 , wherein the Clostridium beijernickii Adh polynucleotide is a heterologous Adh polynucleotide from Clostridium beijernickii NCIMB 8052.

10. A method of producing butanol comprising culturing the recombinant solventogenic microorganism of claim 1 , such that butanol is produced.

11. A method for making the recombinant solventogenic microorganism of claim 1 , comprising transforming a solventogenic microorganism with:
(a) a nucleic acid molecule comprising at least one Clostridium beijerinckii Adh polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 14, wherein the polynucleotide encodes a polypeptide having alcohol dehydrogenase activity,
(b) a nucleic acid molecule comprising at least one Clostridium beijerinckii Bcd polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 2, wherein the polynucleotide encodes a polypeptide having butyryl-CoA dehydrogenase activity, and
(c) a nucleic acid molecule comprising at least one Clostridium beijerinckii Buk polynucleotide comprising a nucleotide sequence having at least 95% sequence identity to the sequence of SEQ ID NO: 3, wherein the polynucleotide encodes a polypeptide having butyrate kinase activity;
wherein the recombinant solventogenic microorganism is capable of (i) more efficient butanol production, (ii) faster butanol production, and/or (iii) increased butanol production relative to the microorganism prior to the transformation with the nucleic acid molecules of (a), (b), and (c).

12. The method claim 11 , wherein, the recombinant solventogenic microorganism is Clostridium sp., Escherichia sp., Clostridium beijerinckii, Clostridium beijerinckii 8052, Clostridium beijernickii BA101, or Clostridium acetobutylicum.

13. The method of claim 11 , wherein the Clostridium beijerinckii Adh polynucleotide is operably linked to a promoter.

14. The recombinant solventogenic microorganism of claim 1 , wherein the nucleic acid molecule comprising at least one Clostridium beijerinckii Adh polynucleotide encodes an alcohol dehydrogenase protein comprising the amino acid sequence of SEQ ID NO: 15.

15. The method of claim 11 , wherein the nucleic acid molecule comprising at least one Clostridium beijerinckii Adh polynucleotide encodes an alcohol dehydrogenase protein comprising the amino acid sequence of SEQ ID NO: 15.

16. The recombinant solventogenic microorganism of claim 1 , which is non-cellulolytic.

Assignees

• Univ Illinois
• Eastman Renewable Mat Llc

Inventors

• Blaschek Hans P
• Stoddard Steven F
• Shi Zhen

Classifications

• C12N9/001

  acting on the CH-CH group of donors (1.3) · CPC title

• C12P7/16

  Butanols · CPC title

• Y02E50/17

  Cross-Sectional Technologies · mapped topic

• C12R1/145

  Chemistry & Metallurgy · mapped topic

• Y02E50/10

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