高等学校化学学报

高等学校化学学报

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超嗜热酯酶APE1547中特殊位置氢键对酶活力和热稳定性的影响

毕云枫1, 解桂秋1,2, 高仁钧1, 鲁静3, 曹淑桂1   

    1. 吉林大学分子酶学工程教育部重点实验室,
    2. 药学院, 长春 130021;
    3. 内蒙古农业大学生物工程学院, 呼和浩特 010018
  • 收稿日期:2006-12-25 修回日期:1900-01-01 出版日期:2007-10-10 发布日期:2007-10-10
  • 通讯作者: 曹淑桂

Effect of Specific Hydrogen Bond on Activity and Thermostability of Hyperthermophilic Esterase APE1547

BI Yun-Feng1, XIE Gui-Qiu1,2, GAO Ren-Jun1, LU Jing3, CAO Shu-Gui1*   

    1. Key Laboratory for Molecular Enzymology and Engineering, Ministry of Education,
    2. School of Pharmaceutical Sciences, Jilin University, Changchun 130021, China;
    3. College of Bioengineering, Inner Mogolia Agricultural University, Hohhot 010018, China
  • Received:2006-12-25 Revised:1900-01-01 Online:2007-10-10 Published:2007-10-10
  • Contact: CAO Shu-Gui

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被引次数

3

摘要： 探讨了APE1547蛋白的β-推进器结构中第3和第4“叶片”间的侧链氢键(Thr127-Gly154, Leu182-Arg145-Glu122)对蛋白质的作用.

关键词: 酯酶APE1547, 氢键, 突变, 活力, 稳定性

Abstract: Thermophilic esterase APE1547 from Aeropyrum pernix K1 contained a β-propeller with seven blades. There are three hydrogen bonding interactions(Thr127-Gly154,Leu182- Arg145-Glu122) between blades 3 and 4 in the β-propeller domain of the APE1547. To examine the role of these hydrogen bonds, we eliminated these three hydrogen bonds between blades 3 and 4, by site-directed mutagenesis. The analysis results of kinetics, thermostability and differential scanning calorimetry(DSC) of the mutants show that Kcat/Km value of each mutation increased, and stability decreased dramatically than wild-type protein. These results strongly suggest that the three specific hydrogen bonds played an important role on maintaining the stability and activity of the esterase APE1547.

Key words: Esterase APE1547, Hydrogen bond, Mutation, Activity, Stability

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