Means and methods for itaconic acid production
AssigneeRheinisch-Westfaelische Technische Hochschule (Rwth) Aachen
InventorsMichael Boelker, Wolfgang Buckel, Sandra Hartmann, Lars M. Blank, Nick Wierckx, Elena Geiser
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Abstract
The present invention relates to a method of producing itaconic acid. Further the present invention relates to nucleic acids encoding an aconitate-delta-isomerase (ADI) and trans-aconitate decarboxylase (TAD) and uses of such nucleic acids. Provided is additionally a recombinant host cell engineered to overexpress nucleic acids of the present invention. Furthermore an expression cassette and a vector are provided which include the respective nucleic acid.
Claims26
A host cell genetically engineered to overexpress at least one polynucleotide sequence selected from one of (a) and (b):
- (a) a polynucleotide sequence selected from the group consisting of a polynucleotide sequence having at least 85% sequence identity with SEQ ID NO:
- 1, a polynucleotide sequence having at least 85% sequence identity with SEQ ID NO:7, a polynucleotide sequence encoding a protein having at least 85% sequence identity with SEQ ID NO:2, and a polynucleotide sequence encoding a protein having at least 85% sequence identity with SEQ ID NO:10, said polynucleotide sequence encoding a protein having aconitate-delta-isomerase (ADI) activity;
- and
- (b) a polynucleotide sequence selected from the group consisting of a polynucleotide sequence having at least 85% sequence identity with SEQ ID NO:
- 6, and a polynucleotide sequence encoding a protein having at least 85% sequence identity with SEQ ID NO:
- 13, said polynucleotide sequence encoding a protein having trans-aconitate decarboxylase (TAD) activity;
- in comparison to a comparable host cell which has not been genetically altered, wherein the host cell is a cell of a fungus or yeast.
A method of producing itaconic acid, comprising
- (a) culturing a recombinant host cell which is genetically engineered to overexpress at least one polynucleotide sequence selected from one of (i) and (ii):
- (i) a polynucleotide sequence selected from the group consisting of a polynucleotide sequence having at least 85% sequence identity with SEQ ID NO:
- 1, a polynucleotide sequence having at least 85% sequence identity with SEQ ID NO:7, a polynucleotide sequence encoding a protein having at least 85% sequence identity with SEQ ID NO:2, and a polynucleotide sequence encoding a protein having at least 85% sequence identity with SEQ ID NO:10, said polynucleotide sequence encoding a protein or fragment thereof having aconitate-delta-isomerase (ADI) activity;
- and
- (ii) a polynucleotide sequence selected from the group consisting of a polynucleotide sequence having at least 85% sequence identity with SEQ ID NO:
- 6, and a polynucleotide sequence encoding a protein having at least 85% sequence identity with SEQ ID NO:
- 13, said polynucleotide sequence encoding a protein or fragment thereof having trans-aconitate decarboxylase (TAD) activity,
- in comparison to a comparable host cell which has not been genetically altered under conditions to allow said host cell to overexpress said polynucleotide sequence (i) or (ii), thereby said host cell is capable of converting cis-aconitate via trans-aconitate to itaconic acid;
- and
- (b) obtaining itaconic acid, wherein
- the host cell is a cell of a fungus or yeast.
Dependencies are inferred from the claim text.