Practical enzymology
Başlık:
Practical enzymology
Yazar:
Bisswanger, Hans.
ISBN:
9783527304448
Ek Yazar:
Yayım Bilgisi:
Weinheim : Wiley-VCH, 2007.
Fiziksel Tanım:
xv, 255 p. : ill. ; 24 cm.
Contents:
Enzyme nomenclature -- Enzyme reactions -- Binding measurements -- Enzymes in technical applications.
Mevcut:*
Library | Materyal Türü | Barkod | Yer Numarası | Durum |
|---|---|---|---|---|
Searching... Pamukkale Merkez Kütüphanesi | Kitap | 0039407 | QP601.B4886 2007 | Searching... Unknown |
Bound With These Titles
On Order
Özet
Özet
Providing a broad experimental, 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.
Essential reading for all scientists and practitioners working with enzymes.
Reviews (1)
Choice Review
This revised work (1st ed., 2004) by Bisswanger (Univ. of Tubingen, Germany) is exactly what the title says, a practical guide to the use and study of enzyme-catalyzed reactions. While an enzymologist would find the book lacking the details and nuances of enzymology, novices to the field will find the book invaluable. Two chapters constitute most of the content. The first gives a background on many aspects of enzyme analysis (e.g., assays, kinetics, inhibition, buffers, pH dependency, analytical methods, sources of error). The second provides protocols for various enzyme assays, covering multiple examples of oxidoreductases, transferases, hydrolases, lyases, isomerases, and ligases. Each protocol is systematically broken into three parts: "Assay Solutions," "Assay Mixture," and "Procedure." One should be able to complete almost any enzyme study by modifying the protocols included. Chapter 2 should provide the knowledge needed to make the necessary modifications. Whether as part of a course or as an independent study, the clearly written text, along with the animated figures provided on a companion website, is an excellent learning tool. This book would be a great resource to both undergraduate and graduate students conducting research that in any way involves enzyme-catalyzed reactions. Summing Up: Highly recommended. Upper-division undergraduates through professionals. L. J. Liotta Stonehill College
Table of Contents
| 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 NADþ 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 ¿-Oxoglutarate Dehydrogenase Complex (OGDHC) |
| 3.7.6.1 Overall Activity by NADþ Reduction |
| 3.7.6.2 ¿-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 ¿-Amylase |
| 3.7.8.8 Amyloglucosidase |
| 3.7.8.9 Lysozyme |
| 3.7.8.10 ¿-Glucosidase |
| 3.7.8.11 ß-Galactosidase |
| 3.7.8.12 ß-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 |
Select a list
The following items were successfully added.
There was an error while adding the following items. Please try again.
One or more items could not be added because you are not logged in.