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| 油包水微乳液中抗体酶催化布洛芬酯选择性水解的酶学特性 | |
| 引用本文： | 杨根生,戚映丹,欧志敏,姚善泾.油包水微乳液中抗体酶催化布洛芬酯选择性水解的酶学特性[J].生物化学与生物物理进展,2009,36(2):182-189. |
| 作者姓名： | 杨根生 戚映丹 欧志敏 姚善泾 |
| 作者单位： | 1. 浙江人学牛物与化学工程与生物工程学系,杭州,310027;浙江工业人学药学院,杭州,310032 2. 浙江工业人学药学院,杭州,310032 3. 浙江人学牛物与化学工程与生物工程学系,杭州,310027 |
| 摘 要： | 根据过渡态理论设计和合成了能诱导产生催化选择性水解布洛芬甲酯的催化抗体的四面体硫酸盐半抗原,并与牛血清白蛋白(BSA)偶联制备成免疫源,通过免疫手段成功筛选出具有加速选择性水解生成S-布洛芬的特异性催化抗体.其Kcat,app/Kuncat,app达1.6x104.进一步地将催化抗体运用到W/O微乳体系(反胶束)中进行布洛芬酯的选择性水解研究,其动力学研究证明其催化过程同样遵循Michaelis.Menten方程.考察了pH值和温度对催化初速度影响,Wo(体系中水和琥珀酸二辛酯磺酸钠(AOT)的摩尔比)对催化初速度影响呈现为钟罩型,最适的Wo.为21. |
| 关 键 词： | 催化抗体 选择性水解 布洛芬 W/O微乳 酶学 琥珀酸二辛酯磺酸钠 |
| 收稿时间： | 2008/5/12 0:00:00 |
| 修稿时间： | 8/4/2008 12:00:00 AM |
Enzymological Characteristics of Catalytic Antibody-catalyzed Enantioselective Hydrolysis of Ibuprofen Ester in Water-in-oil Microemulsion |
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| YANG Gen-Sheng,QI Ying-Dan,OU Zhi-Min and YAO Shan-Jing.Enzymological Characteristics of Catalytic Antibody-catalyzed Enantioselective Hydrolysis of Ibuprofen Ester in Water-in-oil Microemulsion[J].Progress In Biochemistry and Biophysics,2009,36(2):182-189. | |
| Authors: | YANG Gen-Sheng QI Ying-Dan OU Zhi-Min and YAO Shan-Jing |
| Institution: | Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China;College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China;College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China;College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou 310032, China;Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027, China |
| Abstract: | The asymmetric hydrolyzation of racemic ibuprofen ester is one of the most important methods for chiral separation of ibuprofen. A catalytic antibody that accelerates the rate of enantioselective hydrolysis of ibuprofen methyl ester was successfully elicited against an immunogen consisting of tetrahedral sulfate hapten attached to bovine serum albumin (BSA). The rate constant enhancement factor Kcat/Kuncat was about 1.6×104. The catalytic activity of the catalytic antibody in a reverse micelle reaction system based on sodium bis (2-ethylhexyl) sodium sulfosuccinate (AOT) in isooctane was studied. Kinetic analysis of the catalytic antibody-catalyzed reaction was found to be possible in this system. Kinetic studies showed that hydrolysis in the microemulsion system follow Michaelis-Menten kinetics. The catalytic antibody can also accelerate catalysis of S-ibuprofen methyl ester in the microemulsion system. Temperature effects, the pH profile, Km,app and Kcat were determined. The dependence of the catalytic antibody hydrolytic activity on the wo (molar ratio of water to surfactant) showed a bell-shaped curve, presenting a maximum at about wo = 21. |
| Keywords: | catalytic antibody enantiosclectivc hydrolysis ibuprofen W/O microemulsion enzymological characteristics bis(2-ethylhexyl) sodium sulfosuccinate |
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