The search for a perfect model to mimic the properties of the selenoenzyme glutathione peroxidase (GPx) has inspired great interest. Rational design and redesign of the structure–function relationship has become an indispensable technique. In this report, the active site of selenosubtilisin was successfully rebuilt by transferring the catalytically essential residue selenocysteine (Sec) to the edge of the substrate-binding pocket of the enzyme by artificial manipulation. Founding on computer-aided molecular simulation, the amino acid residue at position 63 (Ser in the wild-type enzyme) was selectively replaced with Sec using a cysteine auxotrophic expression system. The novel seleno⁶³-subtilisin E gave a prominent 100-fold higher efficiency than the original seleno²²¹-subtilisin E for GPx activity. Moreover, this seleno⁶³-subtilisin E also had efficient hydrolase activity.

寻找一个完美的模型来模拟硒代酶谷胱甘肽过氧化物酶（GPx）的特性引起了人们极大的兴趣。结构-功能关系的合理设计和重新设计已成为必不可少的技术。在此报告中，通过人工操作将催化必需残基硒代半胱氨酸（Sec）转移到酶的底物结合口袋边缘，成功地重建了硒代枯草杆菌蛋白酶的活性位点。在计算机辅助分子模拟的基础上，使用半胱氨酸营养缺陷型表达系统用Sec选择性取代了位置63（野生型酶中的Ser）的氨基酸残基。新型seleno ^63-枯草杆菌蛋白酶E的效率比原始seleno ²²¹高100倍-枯草杆菌蛋白酶E用于GPx活性。而且，这种硒代^63-枯草杆菌蛋白酶E也具有有效的水解酶活性。