| Description |
xv, 255 pages : illustrations ; 24 cm |
| Contents |
Machine derived contents note: Abbreviations. -- 1 Introduction. -- 2 Enzyme Nomenclature. -- 3 Enzyme Reactions. -- 3.1 Theory of the Enzyme Reaction. -- 3.1.1 Reaction Order. -- 3.1.1.1 Zero-order Reactions. -- 3.1.1.2 First-order Reactions. -- 3.1.1.3 Second- and Higher-order Reactions. -- 3.2 Michaelis?Menten Equation. -- 3.3 Theory of the Enzyme Assay. -- 3.3.1 Analysis of Progress Curves. -- 3.3.2 How To Develop an Enzyme Assay. -- 3.3.2.1 General Considerations. -- 3.3.2.2 pH. -- 3.3.2.3 Ionic Strength and Buffer. -- 3.3.2.4 Temperature. -- 3.3.2.5 Special Components. -- 3.3.2.6 Concentration of Components and Technical Implications. -- 3.3.3 Enzyme Activity. -- 3.3.3.1 Definition of Enzyme Units. -- 3.3.3.2 Absorption Coefficient. -- 3.3.3.3 Calculation of Enzyme Activity. -- 3.4 Theory of Coupled Enzyme Reactions. -- 3.4.1 Two Coupled Reactions. -- 3.4.2 Three Coupled Enzyme Reactions. -- 3.5 Substrate Determination. -- 3.5.1 End-point Method. -- 3.5.2 Coupled Enzyme Reactions. -- 3.5.3 Kinetic Method for Substrate Determination. -- 3.5.4 Enzymatic Cycling. -- 3.6 Instrumental Aspects. -- 3.6.1 Spectroscopic Methods. -- 3.6.1.1 UV/Visible Photometry. -- 3.6.1.2 Turbidity Measurements. -- 3.6.1.3 Fluorimeter. -- 3.6.1.4 Luminometry. -- 3.6.1.5 Polarimetry. -- 3.6.2 Electrochemical Measurements. -- 3.6.2.1 pH Meter. -- 3.6.2.2 pH Stat. -- 3.6.2.3 Potentiometry. -- 3.6.3 Analysis of Reactions that Release or Consume Gas. -- 3.6.3.1 Warburg Manometer. -- 3.6.3.2 Radioactive Labeling. -- 3.6.3.3 Oxygen and Carbon Dioxide Electrodes. -- 3.7 Enzyme Assays. -- 3.7.1 General Considerations. -- 3.7.2 Practical Aspects. -- 3.7.3 Buffers and Solutions. -- 3.7.3.1 Theoretical Aspects. -- 3.7.3.2 Preparation of Buffers. -- 3.7.3.3 Frequently Used Buffers and Solutions. -- 3.7.3.4 Calculation of Enzyme Activities. -- 3.7.4 Oxidoreductase Assays. -- 3.7.4.1 Optical Assays. -- 3.7.4.2 Fluorimetric Assays. -- 3.7.4.3 Alcohol Dehydrogenase. -- 3.7.4.4 Shikimate Dehydrogenase. -- 3.7.4.5 l-Lactate Dehydrogenase. -- 3.7.4.6 Isocitrate Dehydrogenase. -- 3.7.4.7 Glucose-6-phosphate Dehydrogenase. -- 3.7.4.8 Malate Dehydrogenase. -- 3.7.4.9 Glucose Oxidase. -- 3.7.4.10 Glyceraldehyde-3-phosphate Dehydrogenase. -- 3.7.4.11 Pyruvate Ferredoxin Oxidoreductase. -- 3.7.4.12 Glutamate Dehydrogenase. -- 3.7.4.13 l-Amino Acid Oxidase. -- 3.7.4.14 Uricase. -- 3.7.4.15 Catalase. -- 3.7.4.16 Peroxidase. -- 3.7.4.17 Luciferase. -- 3.7.5 Pyruvate Dehydrogenase Complex (PDHC). -- 3.7.5.1 Overall Activity of PDHC by NAD ̓Reduction. -- 3.7.5.2 Overall Activity of PDHC by Dismutation Assay. -- 3.7.5.3 Pyruvate Dehydrogenase (Lipoamide). -- 3.7.5.4 Dihydrolipoamide Acetyltransferase. -- 3.7.5.5 Dihydrolipoamide Dehydrogenase. -- 3.7.6 [alpha]-Oxoglutarate Dehydrogenase Complex (OGDHC). -- 3.7.6.1 Overall Activity by NAD ̓Reduction. -- 3.7.6.2 [alpha]-Oxoglutarate Dehydrogenase. -- 3.7.7 Transferases. -- 3.7.7.1 Fatty Acid Synthase. -- 3.7.7.2 Phosphorylase a. -- 3.7.7.3 Hexokinase. -- 3.7.7.4 Pyruvate Kinase. -- 3.7.7.5 Acetate Kinase. -- 3.7.7.6 3-Phosphoglycerate Kinase. -- 3.7.8 Hydrolases. -- 3.7.8.1 Lipase. -- 3.7.8.2 Cholinesterase. -- 3.7.8.3 Acetylcholinesterase. -- 3.7.8.4 Alkaline Phosphatase. -- 3.7.8.5 Acid Phosphatase. -- 3.7.8.6 Ribonuclease (Pancreatic). -- 3.7.8.7 [alpha]-Amylase. -- 3.7.8.8 Amyloglucosidase. -- 3.7.8.9 Lysozyme. -- 3.7.8.10 [alpha]-Glucosidase. -- 3.7.8.11 [beta]-Galactosidase. -- 3.7.8.12 [beta]-Fructosidase. -- 3.7.9 Proteases. -- 3.7.9.1 Anson Assay. -- 3.7.9.2 Casein Assay. -- 3.7.9.3 Azocasein Assay. -- 3.7.9.4 Ninhydrin Assay. -- 3.7.9.5 Leucine Aminopeptidase. -- 3.7.9.6 Chymotrypsin. -- 3.7.9.7 Pepsin. -- 3.7.9.8 Trypsin. -- 3.7.9.9 Asparaginase. -- 3.7.9.10 Urease. -- 3.7.9.11 Adenosine Triphosphatase. -- 3.7.10 Lyases. -- 3.7.10.1 Pyruvate decarboxylase. -- 3.7.10.2 Aldolase. -- 3.7.10.3 Anthranilate Synthase. -- 3.7.10.4 Carbonic Anhydrase. -- 3.7.10.5 Fumarase. -- 3.7.11 Isomerases. -- 3.7.11.1 Glucose/Xylose isomerase. -- 3.7.12 Determination of Nicotinamide Nucleotides by Enzymatic Cycling. -- 3.7.12.1 Determination of NADP(H). -- 3.7.12.2 Determination of NAD(H). -- 3.8 Diverse Methods. -- 3.8.1 Protein Determination. -- 3.8.1.1 Biuret Method. -- 3.8.1.2 BCA Assay. -- 3.8.1.3 Lowry Assay. -- 3.8.1.4 Coomassie Binding Assay (Bradford Assay). -- 3.8.1.5 Absorption Method. -- 3.8.1.6 Fluorimetric Assay. -- 3.8.1.7 Ninhydrin Assay. -- 3.8.1.8 Modified Ninhydrin Assay for Protein Determination without Hydrolysis. -- 3.8.1.9 Protein Determination with 2-Hydroxy-1-naphthaldehyde. -- 3.8.2 Phosphate Determination. -- 3.8.3 Concentration of Enzyme Solutions. -- 3.8.3.1 Precipitation. -- 3.8.3.2 Ultrafiltration and Dialysis. -- 3.8.3.3 Ultracentrifugation. -- 3.8.3.4 Lyophilization. -- 3.8.3.5 Miscellaneous Techniques. -- 3.9 Enzyme Immunoassays. -- 3.9.1 Non-competitive Solid-phase Enzyme Immunoassay. -- 3.9.2 Competitive, Solid-phase Enzyme Immunoassay. -- 3.9.3 Methods for Enzyme Immunoassays and Immobilization Techniques. -- 3.9.3.1 Protein Coupling to Cyanogen Bromide-activated Agarose. -- 3.9.3.2 Coupling of Diaminohexyl Spacer. -- 3.9.3.3 Periodate Activation of Cellulose. -- 3.9.3.4 Conjugation of Proteins (Antibodies) to Enzymes (Peroxidase). -- 3.9.3.5 Introduction of Thiol Groups into Antibodies or Proteins. -- 3.9.3.6 Conjugation of b-galactosidase to Antibodies by MBS. -- 3.9.3.7 Conjugation of Alkaline Phosphatase to Antibodies by Glutaraldehyde. -- 3.9.3.8 Cross-linking of Proteins with Dimethyl Suberimidate. -- 4 Binding Measurements. -- 4.1 Irreversible, Reversible, Specific, and Unspecific Binding. -- 4.1.1 General Considerations. -- 4.1.2 Experimental Aspects. -- 4.1.2.1 Amount of Enzyme or Macromolecule. -- 4.2.1.2 Stability of the Enzyme or Macromolecule. -- 4.1.2.3 Stability of the Ligand. -- 4.1.2.4 Concentration of Components. -- 4.2 Binding Measurement by Size Discrimination. -- 4.2.1 Ultrafiltration. -- 4.2.2 Equilibrium Dialysis. -- 4.2.2.1 Determination of Indole Binding to Bovine Serum Albumin. -- 4.2.3 Evaluation of Binding Experiments. -- 4.2.4 Gel Filtration. -- 4.2.5 Ultracentrifugation. -- 4.3 Spectroscopic Techniques. -- 4.3.1 Difference Spectroscopy. -- 4.3.1.1 Difference Spectroscopic Determination of Binding of Ligands to Catalase. -- 4.3.1.2 Evaluation of Spectroscopic Binding Curves. -- 4.3.2 Fluorescence Spectroscopy. -- 4.3.2.1 ANS Binding to Bovine Serum Albumin. -- 4.4 Other Methods for Binding Measurements. -- 4.4.1 Radioactive Labeling. -- 4.4.2 Reflectrometric Interference Spectroscopy. -- 5 Enzymes in Technical Applications. -- 5.1 Principles of Enzyme Immobilization. -- 5.1.1 Adsorption. -- 5.1.2 Entrapment. -- 5.1.3 Encapsulation. -- 5.1.4 Cross-linking. -- 5.1.5 Covalent Binding to Solid Supports. -- 5.1.5.1 Supports. -- 5.1.5.2 Spacer. -- 5.2 Methods for Enzyme Immobilization. -- 5.2.1 Microencapsulation inside Nylon Beads. -- 5.2.2 Entrapment in Polyacrylamide. -- 5.2.3 Covalent Immobilization of Enzymes on Nonporous Glass Surfaces. -- 5.2.4 Immobilization on Controlled-pore Glass. -- 5.2.5 Covalent Immobilization of Enzymes to Polyamide. -- 5.2.6 O-Alkylation with Triethyloxoniumtetrafluoroborate. -- 5.2.7 Immobilization of Enzymes to Amine Groups after Partial Hydrolysis of Polyamide. -- 5.2.8 Immobilization of Enzymes to Carboxyl Groups after Partial Hydrolysis of Polyamide. -- 5.2.9 Immobilization of Enzymes to Polyester. -- 5.2.10 Immobilization by Alkaline Hydrolysis and Activation with Tosylchloride. -- 5.2.11 Alkaline Hydrolysis and Activation by Carbonyldiimidazol. -- 5.3 Analysis of Immobilized Enzymes. -- 5.3.1 Protein Assays. -- 5.3.2 Enzyme Assays. -- 5.3.2.1 Modified Optical Assay for Immobilized Enzymes. -- 5.3.2.2 Cofactors in Reactions with Immobilized Enzymes. -- 5.4 Enzyme Reactors. -- 5.4.1 Batch Reactor (Stirred-tank Reactor). -- 5.4.2 Membrane Reactor. -- 5.4.3 Solid-bed Reactor. -- 5.4.4 Immobilized Cells. -- 5.5 Biosensors. -- 5.5.1 Enzyme Electrodes. -- 5.5.2 Immunoelectrodes. -- 5.5.3 Other Biosensors. -- 5.5.4 Bioaffinity Sensors. -- 5.6 Immobilized Enzymes in Therapy. -- Index |
| Summary |
Providing a broad experimental approach, this book contains a theoretical introduction together with practical protocols, considering all aspects of enzymology. The fundamental experiments in enzymology are presented in very clear and easily realizable protocols, with each experiment accompanied by a section giving the theoretical background. |
| Notes |
Formerly CIP. Uk |
| Bibliography |
Includes bibliographical references and index |
| Notes |
Translated into English |
| Subject |
Enzymology.
|
|
Enzymes.
|
| LC no. |
2004301947 |
| ISBN |
3527304444 |
|
