Enzyme assays : high-throughput screening, genetic selection, and fingerprinting / / edited by Jean-Louis Reymond

• Pubblicazione: Weinheim, Germany, : Wiley-VCH, c2006
• Descrizione fisica: 1 online resource (388 p.)
• Disciplina: 572.76
• Soggetto topico: Enzymes - Analysis
• Altri autori: ReymondJean-Louis
• Note generali: Description based upon print version of record.
• Nota di bibliografia: Includes bibliographical references and index.
• Nota di contenuto: Enzyme Assays; Contents; Preface; List of Contributors; Introduction; Enzyme Assays; Part I: High-throughput Screening; Part II: Genetic Selection; Part III: Enzyme Fingerprinting; Enzyme Assays in Other Areas; How to Use this Book; Part I High-throughput Screening; 1 Quantitative Assay of Hydrolases for Activity and Selectivity Using Color Changes; 1.1 Overview; 1.2 Direct Assays Using Chromogenic Substrates; 1.3 Indirect Assays Using Coupled Reactions - pH Indicators; 1.3.1 Overview of Quantitative Use of pH Indicator Assay; 1.3.2 Applications ; 1.3.2.1 Searching for an Active Hydrolase (Testing Many Hydrolases Toward One Substrate)1.3.2.2 Substrate Mapping of New Hydrolases (Testing Many Substrates Toward Hydrolase); 1.3.3 Comparison with Other Methods; 1.4 Estimating and Measuring Selectivity; 1.4.1 Estimating Selectivity without a Reference Compound; 1.4.2 Quantitative Measure of Selectivity Using a Reference Compound (Quick E and Related Methods); 1.4.2.1 Chromogenic Substrate; 1.4.2.2 pH Indicators; 1.4.3 Application; 1.4.3.1 Substrate Mapping of Hydrolases; 1.4.3.2 Screening of Mutants in Directed Evolution ; 1.4.4 Advantages and DisadvantagesReferences; 2 High-throughput Screening Systems for Assaying the Enantioselectivity of Enzymes; 2.1 Introduction; 2.2 UV/Vis Spectroscopy-based Assays; 2.2.1 Assay for Screening Lipases or Esterases in the Kinetic Resolution of Chiral p-Nitrophenyl Esters; 2.2.2 Enzyme-coupled UV/Vis-based Assay for Lipases and Esterases; 2.2.3 Enzymatic Method for Determining Enantiomeric Excess (EMDee); 2.2.4 UV/Vis-based Enzyme Immunoassay as a Means to Measure Enantiomeric Excess; 2.2.5 Other UV/Vis-based ee-Assays; 2.3 Assays Using Fluorescence ; 2.3.1 Umbelliferone-based Systems2.3.2 Fluorescence-based Assay Using DNA Microarrays; 2.3.3 Other Fluorescence-based ee-Assays; 2.4 Assays Based on Mass Spectrometry (MS); 2.4.1 MS-based Assay Using Isotope Labeling; 2.5 Assays Based on Nuclear Magnetic Resonance Spectroscopy; 2.6 Assay Based on Fourier Transform Infrared Spectroscopy for Assaying Lipases or Esterases; 2.7 Assays Based on Gas Chromatography; 2.8 Assays Based on HPLC; 2.9 Assays Based on Capillary Array Electrophoresis; 2.10 Assays Based on Circular Dichroism (CD) ; 2.11 Assay Based on Surface-enhanced Resonance Raman Scattering2.12 Conclusions; References; 3 High-throughput Screening Methods Developed for Oxidoreductases; 3.1 Introduction; 3.2 High-throughput Methods for Various Oxidoreductases; 3.2.1 Dehydrogenases; 3.2.1.1 Colorimetric Screen Based on NAD(P)H Generation; 3.2.1.2 Screens Based on NAD(P)H Depletion; 3.2.2 Oxidases; 3.2.2.1 Galactose Oxidase; 3.2.2.2 D-Amino Acid Oxidase; 3.2.2.3 Peroxidases; 3.2.3 Oxygenases; 3.2.3.1 Assays Based on Optical Properties of Substrates and Products ; 3.2.3.2 Assays Based on Gibbs' Reagent and 4-Aminoantipyrine
• Sommario/riassunto: Edited by one of the leading experts in the field, this book fills the need for a book presenting the most important methods for high-throughput screenings and functional characterization of enzymes. It adopts an interdisciplinary approach, making it indispensable for all those involved in this expanding field, and reflects the major advances made over the past few years.For biochemists, analytical, organic and catalytic chemists, and biotechnologists.
• Titolo autorizzato: Enzyme assays
• ISBN: 1-280-85428-6; 9786610854288; 3-527-60784-6; 3-527-60721-8
• Formato: Materiale a stampa
• Livello bibliografico: Monografia
• Lingua di pubblicazione: Inglese
• Record Nr.: 9910877202003321
• Lo trovi qui: Univ. Federico II