Quantitative Assay of Hydrolases for Activity and Selectivity Using Color Changes

Originally published in: Enzyme Assays. Edited by Jean-Louis Reymond. Copyright © 2006 Wiley-VCH Verlag GmbH & Co. KGaA Weinheim. Print ISBN: 3-527-31095-1 The sections in this article are Introduction Direct Assays Using Chromogenic Substrates Indirect Assays Using Coupled Reactions – pH Indicators Overview of Quantitative Use of pH Indicator Assay Applications Searching for an Active Hydrolase (Testing Many Hydrolases Toward One Substrate) Substrate Mapping of New Hydrolases (Testing Many Substrates Toward Hydrolase) Comparison with Other Methods Estimating and Measuring Selectivity Estimating Selectivity without a Reference Compound Quantitative Measure of Selectivity Using a Reference Compound (Quick E and Related Methods) Chromogenic Substrate pH Indicators Application Substrate Mapping of Hydrolases Screening of Mutants in Directed Evolution Advantages and Disadvantages Acknowledgments Keywords: enzyme assays; high-throughput screening; quantitative assay of hydrolases for activity and selectivity using color changes; direct assays using chromogenic substrates; indirect assays using coupled reaction; pH indicators; estimating and measuring selectivity; chromogenic substrate; application

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