Recombinant cell producing 2-hydroxyisobutyric acid

Patent metadata

Field	Value
Publication number	US-10174349-B2
Application number	US-200913001204-A
Country	US
Kind code	B2
Filing date	Apr 28, 2009
Priority date	Jun 27, 2008
Publication date	Jan 8, 2019
Grant date	Jan 8, 2019

Abstract

The invention relates to a cell which has been genetically modified so as to be capable of producing more 2-hydroxyisobutyric acid or more polyhydroxyalkanoates containing 2-hydroxyisobutyric acid monomer units than its wild type, characterized in that 2-hydroxyisobutyric acid or polyhydroxyalkanoates containing 2-hydroxyisobutyric acid monomer units are produced via acetoacetyl-coenzyme A as intermediate and 3-hydroxybutyryl-coenzyme A as precursor.

First claim

The invention claimed is:

1. A process tor preparing 2-hydroxyisobutyric acid, the process comprising:
a-1) providing a 3-hydroxybutryrate producing microorganism wherein said microorganism is a mutant of a microorganism which has an activity of an enzyme E 4 which is a polyhydroxyalkanoate synthase capable of catalyzing the conversion of 3-hydroxylbutyryl-coenzyme A to polyhydroxybutyrate wherein said mutant lacks activity of enzyme E 4
a-2) transforming said 3-hydroxybutryrate producing microorganism with a nucleic acid encoding a mutase having at least 95% amino acid sequence identity to the amino acid sequence encoded by the Aquincola tertiaricarbonis icmA and icmB wild type genes to obtain transformed cells comprising overexpressed enzyme E 3 which catalyzes the conversion of 3-hydroxylbutyryl-coenzyme A to 2-hydroxyisobutyryl-coenzyme A,
b) after the transforming in a-2), contacting the transformed cells with a nutrient medium including a carbon source under conditions where 2-hydroxyisobutyric acid is produced from the carbon source; and optionally,
c) purifying the 2-hydroxyisobutyric acid from the nutrient medium,
wherein the 3-hydroxybutryrate producing microorganism is selected from the group consisting of Alcaligenes latus, Bacillus subtilis, Methylobacterium extorquens, Ralstonia eutropha, Rhodospirillum rubrum, Rhodobacter sphaeroides, Paracoccus versutus, Pseudomonas aeruginosa, Pseudomonas putida , and Acinetobacter calcoaceticus.

2. A process for preparing methacrylic acid or a methacrylic ester, comprising:
IA) preparing 2-hydroxyisobutyric acid by the process according to claim 1 , and optionally purifying and/or neutralizing the 2-hydroxyisobutyric acid; and
IB) dehydrating the 2-hydroxyisobutyric acid to obtain methacrylic acid, and optionally esterifying the methacrylic acid to obtain the methacrylic ester.

3. A process for preparing poly(methacrylic) acid or a poly(methacrylic) ester, comprising:
IIIA) preparing methacrylic acid by a process according to claim 2 ; and
IIIB) free-radical polymerizing the methacrylic acid,
wherein carboxyl groups of the methacrylic acid are optionally esterified at least partially prior to or after the free-radical polymerization.

4. The process of claim 1 , wherein a concentration of the 2-hydroxyisobutyric acid produced in the contacting in (b) is up to 44 μg/ml.

5. The process of claim 1 , wherein the contacting in (b) is carried out under aerobic conditions.

6. The process of claim 1 , wherein the process comprises the (c) purifying of the 2-hydroxyisobutyric acid from the nutrient medium.

7. The process of claim 1 , wherein the process does not comprise the (c) purifying of the 2-hydroxyisobutyric acid from the nutrient medium.

8. The process of claim 1 , wherein the 3-hydroxybutryrate producing microorganism is Ralstonia eutropha.

9. The process of claim 1 , further comprising:
introducing air comprising oxygen into the nutrient medium,
wherein the introducing of the air and the contacting in (b) are performed at the same time.

10. The process of claim 1 ,
wherein the contacting in (b) is carried out under aerobic conditions for at least 30 hours.

11. The process of claim 1 , wherein the 3-hydroxybutryrate producing microorganism transformed in a-2) is Ralstonia eutropha PHB-4.

Assignees

• Reinecke Liv
• Schaffer Steffen
• Marx Achim
• Poetter Markus
• Haas Thomas
• Evonik Roehm Gmbh

Inventors

• Reinecke Liv
• Schaffer Steffen
• Marx Achim
• Poetter Markus
• Haas Thomas

Classifications

• C12N9/90 Primary

  Isomerases (5.) · CPC title

• C08F8/14

  Esterification · CPC title

• C07C51/347

  by reactions not involving formation of carboxyl groups · CPC title

• C12P7/625 Primary

  Polyesters of hydroxy carboxylic acids · CPC title

• C12Y504/99002

  Methylmalonyl-CoA mutase (5.4.99.2) · CPC title