Loading patent…
AssigneesZhejiang University of Science and Technology, Enzymaster (Ningbo) Bio-Engineering Co., Ltd
InventorsJUN HUANG, Chunyan Liu, Lehe Mei, Haibin Chen, Changjiang LV, Sheng Hu, Hongpeng Wang, Weirui Zhao, Fangfang Fan, Ye Li, Linka Yu, Yifeng Zhou
Estimating the term…
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.
A ω-transaminase mutant obtained by DNA synthetic shuffling combined mutation, wherein the ω-transaminase mutant is obtained by 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, and the mutation site of the ω-transaminase mutant is any one of:
Dependencies are inferred from the claim text.