ω-Transaminase mutant obtained by DNA synthetic shuffling combined mutation and use
US12152255B2 · kind B2 · utility
Assignees
Inventors
Key dates
| Filing date | Dec 6, 2021 |
| Grant date | Nov 26, 2024 |
| Priority date | — |
| Expiry date | Dec 6, 2041 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY02P20/584
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
The present invention discloses a ω-transaminase mutant obtained through DNA synthetic shuffling combined mutation. The ω-transaminase mutant is obtained through point mutation of a wild type ω-transaminase from Aspergillus terrus. The amino acid sequence of the wild type ω-transaminase is shown in SEQ ID NO: 1. The mutation site of the ω-transaminase mutant is any one of: (1) F115L-H210N-M150C-M280C; (2) F115L-H210N; (3) F115L-H210N-E253A-I295V; (4) I77L-F115L-E133A-H210N-N245D; (5) I77L-Q97E-F115L-L118T-E253A-G292D; (6) I77L-E133A-N245D-G292D; and (7) H210N-N245D-E253A-G292D. According to the present invention, forward mutations obtained in the previous stage are randomly combined through a DNA synthetic shuffling combined mutation method. It is verified through experiments that this method can effectively improve the probability of forward mutation and increase experimental efficiency and feasibility, and is capable of obtaining mutant enzymes with thermodynamic stability remarkably superior to that of wild enzymes via screening.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.