Recombinant cell and method of producing itaconic acid
A recombinant cell and a method of producing itaconic acid using such recombinant cell. The recombinant cell is of the genus Methylorubrum and includes a first polynucleotide sequence as defined in SEQ ID NO: 1 or a homologue thereof operably linked to a regulatory sequence.
1. A recombinant cell comprising a first polynucleotide encoding a protein or fragment thereof having cis-aconitic acid decarboxylase (CAD) activity for producing itaconic acid from a carbon substrate, wherein the first polynucleotide comprises SEQ ID NO: 1 operably linked to a regulatory sequence, and the recombinant cell is of the genus Methylorubrum.
2. The recombinant cell of claim 1 , further comprising a second polynucleotide, wherein the second polynucleotide comprises SEQ ID NO: 2.
3. The recombinant cell of claim 1 , wherein the carbon substrate is selected from the group consisting of methanol, methylamine, formate, pyruvate, succinate, lactate and acetate.
4. The recombinant cell of claim 1 , wherein the carbon substrate is selected from the group consisting of acetate, succinate and methanol.
5. The recombinant cell of claim 1 , wherein the carbon substrate is methanol.
6. The recombinant cell of claim 1 , wherein the recombinant cell is Methylorubrum extorquens.
7. The recombinant cell of claim 1 , wherein the recombinant cell is Methylorubrum extorquens AM1.
8. The recombinant cell of claim 1 , wherein the regulatory sequence is a promoter comprising SEQ ID NO: 3.
9. The recombinant cell of claim 1 , wherein the CAD is one derivable from or found in Aspergillus terreus.
10. A method of producing itaconic acid using a recombinant cell, wherein the recombinant cell comprises a first polynucleotide encoding a protein or a fragment thereof having cis-aconitic acid decarboxylase (CAD) activity for producing itaconic acid from a carbon substrate, wherein the first polynucleotide comprises SEQ ID NO: 1, wherein said method comprises the step of incubating the recombinant cell in a medium containing a carbon substrate for producing itaconic acid.
11. The method of claim 10 , wherein the recombinant cell further comprises a second polynucleotide, wherein the second polynucleotide comprises SEQ ID NO: 2.
12. The method of claim 10 , wherein the carbon substrate is selected from the group consisting of methanol, methylamine, formate, pyruvate, succinate, lactate and acetate.
13. The method of claim 10 , wherein the carbon substrate is selected from the group consisting of acetate, succinate and methanol.
14. The method of claim 10 , wherein the carbon substrate is methanol.
15. The method of claim 10 , wherein the recombinant cell is Methylorubrum extorquens.
16. The method of claim 10 , wherein the recombinant cell is Methylorubrum extorquens AM1.
17. The method of claim 10 , wherein the regulatory sequence is a promoter comprising SEQ ID NO: 3.
18. The method of claim 10 , wherein the CAD is one derivable from or found in Aspergillus terreus.