Nitrilase mutant and application thereof in the synthesis of 1-cyanocyclohexyl acetic acid
US12428656B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Dec 11, 2020 |
| Grant date | Sep 30, 2025 |
| Priority date | — |
| Expiry date | Nov 30, 2042 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12R2001/19
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides a nitrilase mutant and application thereof in the synthesis of 1-cyanocyclohexyl acetic acid, the nitrilase mutant is obtained by mutating one or two of the amino acids at position 180 and 205 of the amino acid sequence shown in SEQ ID No. 2. In the present invention, by semi-rational design and protein molecular modification, the specific enzyme activity of the nitrilase double mutant AcN-G180D/A205C was increased by up to 1.6 folds, and the conversion rate>99%. And the reaction time was shortened to a quarter of the original using the recombinant Escherichia coli containing the nitrilase mutant to hydrolyze 1-cyanocyclohexylacetonitrile at high temperature (50° C.). Therefore, the mutants obtained by the present invention have a good application prospect in efficiently catalyzing 1-cyanocyclohexylacetonitrile to synthesize gabapentin intermediate, 1-cyanocyclohexyl acetic acid.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.