Diketoreductase mutant and application thereof
The application provides a Diketoreductase (DKR) mutant, its nucleotide coding sequence, and an expression cassette, recombinant vector and host cell containing the sequence, as well as a method for application of the mutant to the preparation of 3R,5S-dicarbonyl compound. An ee value of the obtained 3R,5S-dicarbonyl compound is higher than 99%, and a de value is about 90%. The DKR mutant is a key pharmaceutical intermediate, and particularly provides an efficient catalyst for synthesis of a chiral dicarbonyl hexanoic acid chain of a statin drug.
1. A Diketoreductase (DKR) mutant, comprising the amino acid sequence shown as
SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 3.
2. A method for producing a 3R,5S-dihydroxy compound, comprising the following steps: contacting the DKR mutant according to claim 1 with a diketone compound for a period of time, wherein the diketone compound is a compound with general formula I:
where R 1 is selected from an aromatic group, an alkyl group, a naphthenic base, an alkyl-substituted aromatic base, a halogen-substituted aryl, an aromatic heterocycle, a heterocycloalkyl or an alkyl-substituted heterocycloalkyl and R2 is selected from an alkyl group, a naphthenic base, a haloalkyl group or a halogen naphthenic base.
3. The method for producing the 3R,5S-dihydroxy compound according to claim 2 , wherein the diketone compound is selected from 6-benzyloxy-3,5-dioxo-tert-butyl hexanoate, 6-benzyloxy-3,5-dioxo-neopentyl hexanoate, 6-benzyloxy-3,5-dioxo-methyl hexanoate and 6-benzyloxy-3,5-dioxo-ethyl hexanoate.
4. A recombinant vector, comprising a nucleotide sequence encoding the DKR mutant according to claim 1 .
5. The recombinant vector according to claim 4 , wherein the recombinant vector is a recombinant expression vector.
6. The recombinant vector according to claim 4 , wherein the nucleotide sequence comprises a sequence selected from one of SEQ ID NOS: 9-11.
7. A host cell transformed or transfected with the recombinant vector according to claim 4 .