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
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浙江省自然科学基金资助项目(Y404353).


Enzymological Characteristics of Catalytic Antibody-catalyzed Enantioselective Hydrolysis of Ibuprofen Ester in Water-in-oil Microemulsion
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This work was supported by a grant from The Natural Science Foundation of Zhejiang, China (Y404353).

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    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1.6×104.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.

    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.

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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

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  • 收稿日期:2008-05-12
  • 最后修改日期:2008-08-04
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  • 在线发布日期: 2008-08-21
  • 出版日期: 2009-02-20