US 12,410,413 B2
Transaminase mutant and use thereof
Hao Hong, Morrisville, NC (US); Gage James, Morrisville, NC (US); Yi Xiao, Tianjin (CN); Yulei Ma, Tianjin (CN); Na Zhang, Tianjin (CN); Xuecheng Jiao, Tianjin (CN); Huiyan Mu, Tianjin (CN); Yibing Cheng, Tianjin (CN); and Shan Cao, Tianjin (CN)
Assigned to ASYMCHEM LABORATORIES (TIANJIN) CO., LTD., Tianjin (CN)
Appl. No. 17/769,204
Filed by ASYMCHEM LABORATORIES (TIANJIN) CO., LTD., Tianjin (CN)
PCT Filed Oct. 28, 2019, PCT No. PCT/CN2019/113743
§ 371(c)(1), (2) Date Apr. 14, 2022,
PCT Pub. No. WO2021/081713, PCT Pub. Date May 6, 2021.
Prior Publication US 2023/0227797 A1, Jul. 20, 2023
Int. Cl. C12N 9/10 (2006.01); C12P 13/00 (2006.01)
CPC C12N 9/1096 (2013.01) [C12P 13/001 (2013.01)] 20 Claims
 
1. A transaminase mutant having the amino acid sequence of SEQ ID NO: 1 with the exception of a mutation of more amino acids, wherein the mutation of more amino acids is selected from the group consisting of: T7C+S47C+Q380L+V379L, T7C+S47C+Q380L+V379D, T7C+S47C+Q380L+V379A, T7C+S47C+Q380L+V379V, T7C+S47C+Q78C+A330C+Y89A, T7C+S47C+Q78C+A330C+Y89E, T7C+S47C+V137C+G313C+Y89V, T7C+S47C+Q380L+V379L+S156P, T7C+S47C+Q78C+A330C+S156Q, T7C+S47C+K249F+1400P+S156A, T7C+S47C+Q380L+V379L+M166F, T7C+S47C+H369A+C404V+M166A, T7C+S47C+H369A+F22P+M166V, T7C+S47C+H369A+L283K+M166S, T7C+S47C+A33V+A57Y+M166Y, T7C+S47C+A33V+A57V+M166G, T7C+S47C+Q380L+V379L+M166F+S424A, T7C+S47C+A33V+A57V+M166Y+S424G, T7C+S47C+Q380L+V379L+F397K+S424L, T7C+S47C+Q380L+V379L+M166F+R416T, T7C+S47C+S292A+A299P+M166K+R416T, T7C+S47C+S292A+A299P+M166F+R416P, T7C+S47C+S292A+A299P+M166F+R416D, T7C+S47C+Q380L+V379L+M166F+Y168I, T7C+S47C+Q380L+V379L+M166F+Y168M, T7C+S47C+S292A+A299P+M166E+Y168A, T7C+S47C+A334L+F367G+M166S+Y168V, T7C+S47C+Q380L+V379L+M166F+N151Q, T7C+S47C+Q380L+V379L+M166F+N151W, T7C+S47C+A334W+F367Q+M166F+N151S, T7C+S47C+Q380L+V379L+M166F+N151Y, T7C+S47C+Q380L+V379L+M166F+R416D, T7C+S47C+Q380L+V379L+M166F+R416P, T7C+S47C+Q380L+V379L+M166F+R416D+1414Q, T7C+S47C+D436A+R457W+M166F+R416D+R444A, T7C+S47C+D436A+R444P+M166S+R416F+D436V, T7C+S47C+D436A+R444Y+M166S+R416F+G419W, T7C+S47C+Q380L+V379L+M166F+R416D+N151W, T7C+S47C+Q380L+V379L+M166F+R416D+N151F, T7C+S47C+D436A+R457C+M166E+R416Q+N151A, T7C+S47C+D436A+R457P+M166W+R416F+N151V, T7C+S47C+D436A+V379L+M166F+R416D+N151E, T7C+S47C+Q380L+V379L+M166F+R416D+Y168E, T7C+S47C+D396Y+F397Q+M166F+R416D+Y168M, T7C+S47C+D396P+F397S+M166G+R416S+Y168S, T7C+S47C+Q380L+V379L+M166F+R416D+Y168A, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+Y168M, T7C+S47C+D396G+F397Y+M166F+R416D+N151W+Y168A, T7C+S47C+D396G+F397Y+M166D+R416F+N151W+Y168S, T7C+S47C+D396G+F397Y+M166D+R416D+N151W+Y168E, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+S424C, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+S424A, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+S424V, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409Q, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409V, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409H, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409M, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409T, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409A, T7C+S47C+D396G+F397Y+M166A+R416V+N151W+F409M, T7C+S47C+D396G+F397Y+M166A+R416V+N151W+F409T, T7C+S47C+D396G+F397Y+M166A+R416V+N151W+F409A, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+S424V+F409C, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+S424A+F409Q, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+S424A+F409C, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409H+E171T, T7C+S47C+Q380L+V379L+M166F+R416D+N151W+F409H+E171A, T7C+S47C+C404Q+R405F+M166S+R416D+N151W+S424V+F409C, T7C+S47C+C404Q+R405F+M166E+R416M+N151W+S424A+F409Q, T7C+S47C+C404Q+R405F+M166S+R416A+N151W+S424A+F409C, T7C+S47C+C404Q+R405F+M166T+R416A+N151W+F409H+E171T, or T7C+S47C+C404Q+R405F+M166Y+R416A+N151W+F409H+E171A; and the transaminase mutant has improved stereoselectivity and enhanced activity compared to the wild type transaminase having the amino acid sequence of SEQ ID NO:1.