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| 耐热对硝基苯磷酸酶的酶学性质 | |
| 引用本文： | 冯从经,伏媛媛,季朝能,顾少华,符文俊,谢毅,毛裕民. 耐热对硝基苯磷酸酶的酶学性质[J]. 中国生物化学与分子生物学报, 2004, 20(5): 648-652 |
| 作者姓名： | 冯从经 伏媛媛 季朝能 顾少华 符文俊 谢毅 毛裕民 |
| 作者单位： | 1. 复旦大学生命科学学院遗传学研究所,遗传工程国家重点实验室,上海,200433;中国科学院上海生命科学研究院植物生理生态研究所,上海,200032 2. 上海联合基因科技研究院,上海,200092 3. 复旦大学生命科学学院遗传学研究所,遗传工程国家重点实验室,上海,200433 4. 中国科学院上海生命科学研究院植物生理生态研究所,上海,200032 |
| 基金项目： | 国家自然科学基金项目 (No .3 0 0 70 161)资助~~ |
| 摘 要： | 通过分离纯化和酶学性质测定 ,确定耐热对硝基苯磷酸酶 (p nitrophenylphosphatase)是金属酶蛋白 ,活性部位含有镁离子 ;该酶是中度耐热蛋白质 ,金属离子有助于提高其热稳定性 .该酶的Km 为 3 0 31mmol L ,Vmax为 313 5 μmol·min-1·mg-1;二级结构中大约有 74 5 %α螺旋 ,2 5 5 % β转角 .在pH 7 0～ 12 0范围内相当稳定 .该酶在SDS溶液中稳定性较差 ,在TritonX 10 0溶液中较为稳定 ,在Tween 2 0溶液中很稳定 . |
| 关 键 词： | 耐热对硝基苯磷酸酶 酶学性质 热稳定性 金属离子 二级结构 |
| 收稿时间： | 2004-10-20 |
| 修稿时间： | 2003-10-06 |
Enzymatic Characteristics of Thermostable p-Nitrophenylphosphatase |
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| FENG Cong-jing ),),FU Yuan-yuan ),JI Chao-neng ),GU Shao-hua ),FU Wen-jun ),XIE Yi ),MAO Yu-min ). Enzymatic Characteristics of Thermostable p-Nitrophenylphosphatase[J]. Chinese Journal of Biochemistry and Molecular Biology, 2004, 20(5): 648-652 | |
| Authors: | FENG Cong-jing ) ) FU Yuan-yuan ) JI Chao-neng ) GU Shao-hua ) FU Wen-jun ) XIE Yi ) MAO Yu-min ) |
| Affiliation: | ( 1) State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Sciences, Fudan University, Shanghai 200433,China; 2) Institute of Plant Physiology and Ecology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China; 3) United Gene Holding Group Corporation, LTD, Shanghai 200092,China |
| Abstract: | Thermostable p-nitrophenylphosphatase (TNPPase) had been expressed in E. coli M15, and purified rapidly. It is sensitive to metal chelator and Mg ion is necessary to its activity. TNPPase exhibited a moderate degree of thermostability, and the thermostability of TNPPase increased significantly when treated with 1 mmol/L Mg, Mn, Cu, Ba, Ca, Ni and Zn ions. TNPPase is stable within a wide range of pH (pH 7.0～12.0). It catalyzed the hydrolysis of p-nitrophenylphosphate with a Michaelis constant 3.031 mmol/L and a V max 313.5 μmol·min -1 ·mg -1 . The α-helix percentage and β-turn percentages were 74.5 and 25.5 in the secondary structure of TNPPase, respectively. TNPPase is not stable in SDS solution, but stable in Triton X-100 and Tween 20 solutions. |
| Keywords: | thermostable nitrophenylphosphatase enzymatic characteristics thermostability metal ion secondary structure |
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