Recombinant expression vector applicable to rapid screening for recombinant strain and application
US-12037632-B2 · Jul 16, 2024 · US
US9290772B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-9290772-B2 |
| Application number | US-201314137785-A |
| Country | US |
| Kind code | B2 |
| Filing date | Dec 20, 2013 |
| Priority date | Feb 18, 2008 |
| Publication date | Mar 22, 2016 |
| Grant date | Mar 22, 2016 |
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The invention relates to a nucleic acid sequence encoding an Aspergillus mitochondrial tricarboxylic acid transporter that can be used in the production of itaconic acid in micro-organisms. Preferably said transporter protein is the protein encoded by the nucleic acid which is located on a chromosome segment of A. terreus that also harbors other genes involved in the itaconic acid biosynthesis and the lovastatin biosynthesis. Also, vectors, hosts and transformed micro-organisms are part of the invention.
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The invention claimed is: 1. A host cell which has been modified to contain a heterologous gene encoding a protein that transports di/tricarboxylate from the mitochondrion to the cytosol, wherein said protein has an amino acid sequence at least 95% identical to ATEG — 09970.1 of SEQ ID NO:3. 2. The host cell of claim 1 , wherein said protein is a tricarboxylate transporter. 3. The host cell of claim 1 , wherein said protein transports cis-aconitate, citrate or isocitrate. 4. The host cell of claim 1 , wherein said host cell is from of a citrate producing microorganism. 5. The host cell of claim 1 , wherein said gene comprises 1) a nucleic acid sequence encoding a mitochondrial tricarboxylic acid transporter from A. terreus, A. niger, A. itaconicus, A. nidulans, A. oryzae , or A. fumigates , or 2) a nucleic acid sequence which encodes the amino acid sequence of ATEG — 09970.1 of SEQ ID NO:3. 6. The host cell of claim 1 , wherein a nucleic acid encoding the enzyme cis-aconitic acid decarboxylase (CAD) and/or a nucleic acid encoding a Major Facilitator Superfamily (MFS) transporter is co-introduced. 7. The host cell of claim 4 , wherein the citrate producing micro-organism 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. 8. The host cell of claim 1 , which is of a lovastatin producing microorganism. 9. The host cell of claim 6 , wherein the CAD is encoded by the nucleotide sequence comprised in ATEG — 09971.1 (SEQ ID NO:5). 10. The host cell of claim 6 , wherein the MFS transporter is encoded by the nucleotide sequence comprised in ATEG — 09972.1 (SEQ ID NO:13). 11. The host cell of claim 8 , wherein the lovastatin producing micro-organism is from 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. 12. The host cell of claim 7 , which is of A. terreus or A. niger. 13. The host cell of claim 1 , wherein the encoding nucleotide sequence is expressed from a vector comprising a promoter capable of driving expression of said sequence.
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