Phosphinothricin dehydrogenase mutant, genetically engineered bacterium and one-pot multi-enzyme synchronous directed evolution method
US12152264B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Dec 26, 2020 |
| Grant date | Nov 26, 2024 |
| Priority date | — |
| Expiry date | May 4, 2042 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12R2001/39
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Disclosed are a phosphinothricin dehydrogenase mutant, a recombinant bacterium and a one-pot multi-enzyme synchronous directed evolution method. The phosphinothricin dehydrogenase mutant, with an amino acid sequence as shown in SEQ ID No.1, is obtained by mutating alanine at position 164 to glycine, arginine at position 205 to lysine, and threonine at position 332 to alanine in a phosphinothricin dehydrogenase derived from Pseudomonas fluorescens. The recombinant bacterium is obtained by introducing a gene encoding the phosphinothricin dehydrogenase mutant into a host cell. The host cell can also incorporate a gene encoding a glucose dehydrogenase or a gene encoding a formate dehydrogenase to undergo synchronous directed evolution to achieve double gene overexpression. The one-pot multi-enzyme synchronous directed evolution method of the present invention can screen recombinant bacteria with greatly improved activity. Compared with other catalysis processes such as the transaminase method, the method for preparing L-PPT of the present invention features relatively simple process, high conversion of raw materials of up to 100%, and high stereo selectivity.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.