Polypeptide tag, highly soluble recombinant nitrilase and application thereof in synthesis of pharmaceutical chemicals
US11807883B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Apr 26, 2021 |
| Grant date | Nov 7, 2023 |
| Priority date | — |
| Expiry date | Apr 26, 2041 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12R2001/19
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention provides a polypeptide tag and its application in the synthesis of pharmaceutical chemicals, the recombinant nitrilase was obtained by connecting a polypeptide tag to the N-terminus of the amino acid sequence of the nitrilase; wherein amino acids at both ends of the polypeptide tag are uncharged glycine G, and the rest are a random combination of any one or more of glycine G, histidine H, glutamic acid E, aspartic acid D, lysine K and arginine R; The activity of the recombinant nitrilase in the preparation of 1-cyanocyclohexyl acetic acid is up to 3034.7 U/g dcw, the polypeptide tag significantly improves the soluble expression of nitrilase, and the whole cell catalyst hydrolyzes 1M substrate with the same concentration 30 minutes faster than the mother enzyme. The method provided by the present invention can also be used for the biocatalytic reaction of other pharmaceutical intermediates as the substrate catalyzed by the nitrilase, improving the activity of the whole cell catalyst in reaction, and also improving the solubility of other types of nitrilases and the activity of the corresponding whole cell catalysts.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.