Engineered microorganisms having resistance to ionic liquids

Patent metadata

Field	Value
Publication number	US-9290768-B2
Application number	US-201313935325-A
Country	US
Kind code	B2
Filing date	Jul 3, 2013
Priority date	Jul 3, 2012
Publication date	Mar 22, 2016
Grant date	Mar 22, 2016

Abstract

The present invention provides for a method of genetically modifying microorganisms to enhance resistance to ionic liquids, host cells genetically modified in accordance with the methods, and methods of using the host cells in a reaction comprising biomass that has been pretreated with ionic liquids.

First claim

What is claimed is:

1. A method of increasing the yield from a reaction in which soluble sugars are a source of carbon, the method comprising incubating a microorganism with biomass pretreated with an ionic liquid, wherein the biomass pretreated with an ionic liquid provides a source of soluble sugars as a carbon source, wherein the microorganism comprises a heterologous gene encoding a Major Facilitator Superfamily 1 (MFS 1) polypeptide operably linked to a promoter, wherein said MFS 1 polypeptide has an amino acid sequence with at least 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:2, and wherein said MFS 1 polypeptide facilitates resistance of the microorganism to said ionic liquid as compared to a microorganism without said heterologous gene.

2. The method of claim 1 , wherein said biomass is incubated with said microorganism in a fermentation reaction that produces an alcohol.

3. The method of claim 1 , wherein said MFS 1 polypeptide comprises the amino acid sequence of SEQ ID NO:2.

4. The method of claim 1 , wherein said heterologous gene is operably linked to a tet repressor sequence.

5. The method of claim 4 , wherein said microorganism is genetically modified to express a tet repressor protein that binds to the tet repressor sequence.

6. The method of claim 1 , wherein said microorganism is a bacteria.

7. The method of claim 6 , wherein said bacteria is Escherichia coli.

8. The method of claim 1 , wherein said microorganism is a yeast or a filamentous fungi.

9. The method of claim 1 , wherein said ionic liquid comprises an anion, wherein said anion is Cl— or acetate.

10. The method of claim 1 , wherein said heterologous gene encoding said MFS 1 polypeptide is integrated into the genome of said microorganism.

11. The method of claim 1 , wherein said microorganism is generated by transforming said microorganism with an expression cassette, wherein the said expression cassette comprises said heterologous gene encoding said MFS 1 polypeptide.

Assignees

• L Livermore Nat Security Llc

Inventors

• Ruegg Thomas Lawrence
• Thelen Michael P

Classifications

• C07K14/265 Primary

  Enterobacter (G) · CPC title

• C12N1/22

  Processes using, or culture media containing, cellulose or hydrolysates thereof · CPC title

• C12N15/70 Primary

  Vectors or expression systems specially adapted for E. coli · CPC title

• C12P7/10

  substrate containing cellulosic material · CPC title

• Y02E50/16

  Cross-Sectional Technologies · mapped topic