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  

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

作　　者：毕云枫[1] 解桂秋[1] 高仁钧[1] 鲁静[2] 曹淑桂[1] 

机构地区：[1]吉林大学分子酶学工程教育部重点实验室,长春130021 [2]内蒙古农业大学生物工程学院,呼和浩特010018

出　　处：《高等学校化学学报》2007年第10期1914-1916,共3页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:30400081;30570405和20672045)资助.

摘　　要：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.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 resuits 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.

关 键 词：酯酶APE1547 氢键 突变 活力 稳定性 

分 类 号：O629[理学—有机化学] Q814[理学—化学]