申请/专利权人：凯莱英生命科学技术(天津)有限公司

申请日：2022-06-21

公开（公告）日：2024-03-12

公开（公告）号：CN114875006B

主分类号：C12N9/10

分类号：C12N9/10;C12N15/54;C12N15/70;C12N1/21;C12P13/00;C12P17/12;C12R1/19

优先权：

专利状态码：有效-授权

法律状态：2024.03.12#授权;2022.08.26#实质审查的生效;2022.08.09#公开

摘要：本发明提供了一种转氨酶突变体及手性胺化合物的制备方法。其中，该转氨酶突变体，包括a具有SEQIDNO：1所示的氨基酸序列的蛋白质；或b具有SEQIDNO：2或SEQIDNO：3所示的氨基酸序列的蛋白质；或c在b中的氨基酸序列的如下至少一个位点：Y89、L380、N86、Y85、T91、P83、K90、S417、S424、F301、G164、T452等，经过氨基酸突变且具有转氨酶功能的蛋白质；d与a、b或c中任一项限定的氨基酸序列具有80％以上同源性且具有转氨酶功能的蛋白质。能够解决现有技术中的转氨酶活性低的问题，适用于酶催化领域。

主权项：1.一种转氨酶突变体，其特征在于，所述转氨酶突变体由SEQIDNO：1发生氨基酸突变得到，所述氨基酸突变选自如下任意一种：V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+I311F；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315C+A31V；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+V327S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295M；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295F；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64A；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409Q；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409V；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409N；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409N+T402K；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409V+T402S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409V+T402S+T126C；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315M+A31V+L295Q+V64M+H154S+F409V+T402S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90Y+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315R+A31V+L295Q+V64M+H154S+F409V+T402S+T126C；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315M+A31V+L295Q+V64M+H154S+F409V+T402S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S+T126C；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S+T126C+F364L；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S+T126C+F364L+R77Q；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S+T126C+F364L+N317Y；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64M+H154S+F409V+T402S+T126C+F364L+A433T；V234A+L380A+Y89D+N86H+Y85M+T91I+P83S+K90G+S417I+S424A+F301S+G164S+T452S+M180V+F449L+F320H+Y322T+D315V+A31V+L295Q+V64MH154S+F409V+T402S+T126C+F364L+N317Y。

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