Rational redesign of the active site of selenosubtilisin with strongly enhanced glutathione peroxidase activity
Rational redesign of the active site of selenosubtilisin with strongly enhanced glutathione peroxidase activity
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合理重新设计硒枯草杆菌蛋白酶活性位点，显着增强谷胱甘肽过氧化物酶活性
DOI：
10.1016/j.jcat.2017.12.006
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发表时间：
2018-03
影响因子：
7.3
通讯作者：
Liu Junqiu
中科院分区：
文献类型：
--
作者：
Wang Tingting;Li Jing;Xu Jiayun;Fan Xiaotong;Zhao Linlu;Qiao Shanpeng;Pan Tiezheng;Liu Junqiu
寻找一个完美的模型来模拟硒酶谷胱甘肽过氧化物酶（GPx）的性质激发了极大的兴趣。结构-功能关系的合理设计和再设计已成为不可或缺的技术。在这份报告中，硒枯草杆菌蛋白酶的活性位点被成功地重建通过转移催化必需残基硒半胱氨酸（Sec）的边缘的底物结合口袋的酶的人工操作。基于计算机辅助分子模拟，使用半胱氨酸营养缺陷型表达系统将位置63处的氨基酸残基（野生型酶中的Ser）选择性地替换为Sec。新的硒代63-枯草杆菌蛋白酶E给出了一个显着的100倍的效率比原来的硒代221-枯草杆菌蛋白酶E的GPx活性。此外，该硒63-枯草杆菌蛋白酶E也具有有效的水解酶活性。
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.
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影响因子：
3.8
作者：
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通讯作者：
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作者：
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影响因子：
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作者：
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