Visualization of enzyme-catalyzed reactions using pH indicators:: Rapid screening of hydrolase libraries and estimation of the enantioselectivity

被引：61

作者：

Morís-Varas, F ^{[1
]}

Shah, A ^{[1
]}

Aikens, J ^{[1
]}

Nadkarni, NP ^{[1
]}

Rozzell, JD ^{[1
]}

Demirjian, DC ^{[1
]}

机构：

[1] ThermoGen Inc, Chicago, IL 60612 USA

来源：

BIOORGANIC & MEDICINAL CHEMISTRY | 1999年 / 7卷 / 10期

关键词：

D O I：

10.1016/S0968-0896(99)00149-2

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The use of pH indicators to monitor hydrolase-catalyzed reactions is described. The formation of acid following an enzyme-mediated hydrolysis causes a drop in the pH that can be visualized by a change in the color of the indicator-containing solution. The best indicators are those showing a color transition within the operational pH range of the hydrolases, like bromothymol blue and phenol red. The enantioselectivity of lipases and esterases can be estimated using single isomers under the same conditions and comparing the color turnover for each one. The method has been tested to quickly evaluate the enantioselectivity of a lipase towards a set of ester substrates and applied to the hierarchical screening of a library of thermophilic esterases. (C) 1999 Elsevier Science Ltd. All rights reserved.

引用

页码：2183 / 2188

页数：6

共 49 条

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