Production of itaconic acid

Patent metadata

Field	Value
Publication number	US-9359612-B2
Application number	US-201414147747-A
Country	US
Kind code	B2
Filing date	Jan 6, 2014
Priority date	Mar 5, 2008
Publication date	Jun 7, 2016
Grant date	Jun 7, 2016

Abstract

The invention relates to a nucleic acid sequence encoding an itaconate transporting Major Facilitator Superfamily Transporter (MFST) gene sequence and the protein encoded thereby. Preferably said sequence is the nucleic acid that comprises the sequence of ATEG_09972.1 of Aspergillus terreus or homologues thereof. Overexpression of the protein enhances the production of itaconic acid in micro-organisms.

First claim

The invention claimed is:

1. Recombinant host cells that have been modified to improve production of itaconate,
said cells having been modified only to contain a nucleic acid that encodes protein wherein said encoded protein consists of the amino acid sequence of an itaconate transporter protein that transports itaconate acids from the cytosol to the extracellular medium; and
optionally nucleic acid that encodes protein wherein said encoded protein consists of the amino acid sequence of the enzyme cis aconic acid decarboxylase (CAD), and optionally nucleic acid that encodes protein wherein said encoded protein consists of the amino acid sequence of a protein capable of transporting di/tricarboxylic acids from the mitochondrion,
wherein the itaconate transporter protein has the amino acid sequence of SEQ ID NO:3 or an amino acid sequence at least 95% identical thereto or SEQ ID NO:6, wherein the host cells are heterologous to said nucleic acid.

2. The host cells of claim 1 , wherein nucleic acid encodes a protein having SEQ ID NO:3 or SEQ ID NO:6.

3. The host cells of claim 1 , which are cells of a citrate producing micro-organism.

4. The host cells of claim 3 , wherein the citrate producing microorganism is A. terreus, A. niger, A. itaconicus, A. nidulans, A. oryzae, A. fumigates, Yarrowia lipolytica, Ustilago zeae, Candida sp., Rhodotorula sp., Pseudozyma antarctica, E. coli , or Saccharomyces cerevisiae.

5. The host cells of claim 4 , wherein the citrate producing microorganism is A. terreus or A. niger.

6. The host cells of claim 1 , which are of a lovastatin producing organism.

7. The host cells of claim 6 , wherein the lovastatin producing microorganism is Monascus spp., Penicillium spp., Hypomyces spp., Doratomyces spp., Phoma spp., Eupenicillium spp., Gymnoascus spp., Pichia labacensis, Candida cariosilognicola, Paecilomyces varioti, Scopulariopsis brevicaulis or Trichoderma spp.

8. The host cells of claim 1 , which are also modified with a nucleic acid that encodes protein wherein said encoded protein consists of the amino acid sequence of the enzyme cis aconic acid decarboxylase (CAD), wherein said nucleic acid encoding the enzyme CAD comprises a nucleotide sequence which encodes the amino acid sequence of SEQ ID NO:12.

9. The host cells of claim 1 , which are also modified with a nucleic acid that encodes protein wherein said encoded protein consists of the amino acid sequence of a protein capable of transporting di/tricarboxylic acids from the mitochondrion, wherein the nucleotide sequence encodes a protein of the amino acid sequence SEQ ID NO:14.

10. The host cells of claim 1 , wherein the nucleic acid encoding itaconate transporter comprises SEQ ID NO:2 or a nucleotide sequence at least 95% identical thereto or SEQ ID NO:5.

11. The host cells of claim 10 , wherein the nucleic acid encoding itaconate transporter comprises SEQ ID NO:2 or SEQ ID NO:5.

Assignees

• TNO
• Dutch Dna Biotech B V

Inventors

• Van Der Werf Maria Johanna
• Punt Peter Jan

Classifications

• C12N15/80 Primary

  for fungi · CPC title

• C07K14/38 Primary

  from Aspergillus · CPC title

• C12P7/44

  Polycarboxylic acids · CPC title