Recombinant host cells comprising phosphoketalase

Patent metadata

Field	Value
Publication number	US-10006058-B2
Application number	US-201615375858-A
Country	US
Kind code	B2
Filing date	Dec 12, 2016
Priority date	Jun 18, 2010
Publication date	Jun 26, 2018
Grant date	Jun 26, 2018

Abstract

The present invention is related to recombinant host cells comprising: (i) at least one deletion, mutation, and/or substitution in an endogenous gene encoding a polypeptide that converts pyruvate to acetaldehyde, acetyl-phosphate or acetyl-CoA; and (ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity. The present invention is also related to recombinant host cells further comprising (iii) a heterologous polynucleotide encoding a polypeptide having phosphotransacetylase activity.

First claim

What is claimed is:

1. A method for the production of isobutanol comprising growing a recombinant host cell comprising:
(i) at least one genetic modification in an endogenous gene encoding a polypeptide having pyruvate decarboxylase activity, wherein the at least one genetic modification reduces or eliminates pyruvate decarboxylase activity;
(ii) a heterologous polynucleotide encoding a polypeptide having phosphoketolase activity, wherein the heterologous polynucleotide encoding a polypeptide having phosphoketolase activity is xpk1 from Lactobacillus plantarum , xpkA from Lactobacillus pentosus MD363, or 6-phosphate phosphoketolase from B. lactis;
(iii) at least one genetic modification in an endogenous gene encoding a polypeptide having glycerol-3-phosphate dehydrogenase activity, wherein the at least one genetic modification reduces or eliminates glycerol-3-phosphate dehydrogenase activity; and
(iv) an isobutanol biosynthetic pathway comprising the substrate to product conversions:
(a) pyruvate to acetolactate,
(b) acetolactate to 2,3-dihydroxyisovalerate,
(c) 2,3-dihydroxyisovalerate to 2-ketoisovalerate,
(d) 2-ketoisovalerate to isobutyraldehyde,
(e) isobutyraldehyde to isobutanol;

under conditions wherein isobutanol is produced and optionally recovering isobutanol.

2. The method of claim 1 , wherein the isobutanol is recovered by distillation, azeotropic distillation, liquid-liquid extraction, adsorption, gas stripping, membrane evaporation, pervaporation, vacuum flash, or combinations thereof.

3. The method of claim 1 , wherein the isobutanol is recovered by contacting the fermentation broth with an extractant.

4. The method of claim 3 , wherein the extractant is C 12 to C 22 fatty alcohols, C 12 to C 22 fatty acids, esters of C 12 to C 22 fatty acids, C 12 to C 22 fatty aldehydes, or mixtures thereof.

5. The recombinant host cell of claim 1 , wherein the polypeptide having phosphoketolase activity comprises at least 85% identity to SEQ ID NO: 481.

6. The recombinant host cell of claim 1 , wherein the recombinant host cell is Saccharomyces cerevisiae.

Assignees

• Butamax Advanced Biofuels Llc

Inventors

• Dauner Michael
• Maggio-Hall Lori Ann
• Tomb Jean-Francois

Classifications

• C12Y401/01001

  Pyruvate decarboxylase (4.1.1.1) · CPC title

• C12N9/1029

  transferring groups other than amino-acyl groups (2.3.1) · CPC title

• C12Y401/02009

  Phosphoketolase (4.1.2.9) · CPC title

• C12N9/88

  Lyases (4.) · CPC title

• C12P7/16 Primary

  Butanols · CPC title