PROCESS FOR PRODUCING ITACONIC ACID UNDER ANAEROBIC CONDITIONS

• US 20180016604A1
• Filed: 02/16/2016
• Published: 01/18/2018
• Est. Priority Date: 02/16/2015
• Status: Active Grant

• Current Assignee: Covestro NV (Covertro Deutschland AG)
• Original Assignee: DSM IP Assets BV (DSM-Firmenich AG)
• Inventors: WEUSTHUIS, Ruud, VUORISTO, Kiira, EGGINK, Gerrit, SANDERS, Johan Pieter Marinus
• Granted Patent: US 10,443,077 B2
• Time in Patent Office: Days
• CPC Class Codes: C12N 15/52 Genes encoding for enzymes or proenzymes
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 7/44 Polycarboxylic acids
• CPC Class Codes: C12Y 401/01006 Aconitate decarboxylase (4.1.1.6)

Abstract

The present invention relates to a method for the production of itaconic acid, which method comprises fermenting a recombinant cell capable of producing itaconic acid in a suitable fermentation medium, thereby to produce itaconic acid, wherein: (a) the recombinant cell: (i) overexpresses cis-aconitate decarboxylase; and (ii) overexpresses a part of the citric acid cycle and/or has reduced activity of a native metabolic route to acetate and/or lactate; and, optionally, (b) the fermentation is carried out under anaerobic conditions.

Claims

1. A method for the production of itaconic acid, which method comprises fermenting a recombinant cell capable of producing itaconic acid in a suitable fermentation medium, thereby to produce itaconic acid, wherein:

(a) the recombinant cell;

(i) overexpresses cis-aconitate decarboxylase; and

(ii) overexpresses a part of the citric acid cycle and/or has reduced activity of a native metabolic route to acetate and/or lactate, wherein overexpression of a part of the citric acid cycle is overexpression of a citrate synthase and/or overexpression of an aconitase and wherein reduced activity of a native metabolic route to acetate is reduced activity of a phosphate acetyltransferase and/or wherein reduced activity of a native metabolic route to lactate is reduced activity of a lactate dehydrogenase;

and, optionally,

(b) the fermentation is carried out under anaerobic conditions, wherein fermentation carried out under anaerobic conditions means a fermentation process run in the absence of oxygen or in which substantially no oxygen is consumed or a fermentation process run under oxygen limited, or micro-aerobical, conditions; and

, optionally

(c) wherein glutamate production is repressed, preferably wherein repression of glutamate production is carried out by carrying out the fermentation under nitrogen free conditions.

2. A method according to claim 1, wherein the cis-aconitate decarboxylase is one derivable from or found in Aspergillus terreus.

3. A method according to claim 1 wherein production of itaconic acid is accompanied by either ethanol and formate production or H₂ and CO₂ production.

4. A method according to any one of the preceding claim 1, wherein glutamate production is repressed.

5. A method according to claim 1, wherein the recombinant cell is a prokaryotic or a eukaryotic cell.

6. A method according to claim 5, wherein the cell is a bacterial cell, preferably E. coli, or a yeast cell, preferably S. cerevisiae.

7. A method according to claim 1 which comprises recovery of the itaconic acid.

8. A method for the production of a pharmaceutical, cosmetic, food, feed or chemical product which method comprises:

(a) preparing itaconic acid by a method according to claim 1 and;

(b) preparing the pharmaceutical, cosmetic, food, feed or chemical product using said itaconic acid.

9. A recombinant cell capable of producing itaconic acid, wherein the recombinant cell:

(i) overexpresses cis-aconitate decarboxylase; and

(ii) overexpresses a part of the citric acid cycle and/or has reduced activity of a native metabolic route to acetate and/or lactate.

10. A recombinant cell according to claim 9.

11. A method for the production of itaconic acid, which method comprises fermenting a recombinant cell capable of producing itaconic acid in a suitable fermentation medium, thereby to produce itaconic acid, wherein the recombinant cell is one according to claim 9.

12. A method according to claim 11, wherein the fermentation is carried out under anaerobic conditions.