Practical enzymology:
Gespeichert in:
| 1. Verfasser: | |
|---|---|
| Format: | Buch |
| Sprache: | English |
| Veröffentlicht: |
Weinheim
Wiley-VCH
2004
|
| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | XV, 255 S. graph. Darst. |
| ISBN: | 3527304444 |
Internformat
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Datensatz im Suchindex
| _version_ | 1804130199375183872 |
|---|---|
| adam_text | Contents
Abbreviations X11I
1 Introduction 1
2 Enzyme Nomenclature 5
3 Enzyme Reactions 7
3.1 Theory of the Enzyme Reaction 7
3.1.1 Reaction Order 7
3.1.1.1 Zero order Reactions 8
3.1.1.2 First order Reactions 8
3.1.1.3 Second and Higher order Reactions 9
3.2 Michaelis Menten Equation 10
3.3 Theory of the Enzyme Assay 16
3.3.1 Analysis of Progress Curves 36
3.3.2 How To Develop an Enzyme Assay 21
3.3.2.1 General Considerations 21
3.3.2.2 pH 22
3.3.2.3 Ionic Strength and Buffer 23
3.3.2.4 Temperature 23
3.3.2.5 Special Components 24
3.3.2.6 Concentration of Components and Technical Implications 26
3.3.3 Enzyme Activity 28
3.3.3.1 Definition of Enzyme Units 28
3.3.3.2 Absorption Coefficient 28
3.3.3.3 Calculation of Enzyme Activity 29
3.4 Theory of Coupled Enzyme Reactions 32
3.4.1 Two Coupled Reactions 32
3.4.2 Three Coupled Enzyme Reactions 35
3.5 Substrate Determination 35
3.5.1 End point Method 35
3.5.2 Coupled Enzyme Reactions 37
3.5.3 Kinetic Method for Substrate Determination 38
Practiaä Enzymoiogy. H. Bisswanger
Copyright C 2004 W1LEYVCH Verlag GmbH Co. KGaA. Weinheim
ISBN 3 527 30444 4
VIII I Contents
3.5.4 Enzymatic Cycling 38
3.6 Instrumental Aspects 40
3.6.1 Spectroscopic Methods 41
3.6.1.1 UV/Visible Photometry 41
3.6.1.2 Turbidity Measurements 46
3.6.1.3 Fluorimeter 47
3.6.1.4 Luminometry 50
3.6.1.5 Polarimetry 53
3.6.2 Electrochemical Measurements 52
3.6.2.1 pH Meter 52
3.6.2.2 pH Stat 53
3.6.2.3 Potentiometry 54
3.6.3 Analysis of Reactions that Release or Consume Gas 54
3.6.3.1 Warburg Manometer 54
3.6.3.2 Radioactive Labeling 55
3.6.3.3 Oxygen and Carbon Dioxide Electrodes 55
3.7 Enzyme Assays 56
3.7.1 General Considerations 56
3.7.2 Practical Aspects 59
3.7.3 Buffers and Solutions 59
3.7.3.1 Theoretical Aspects 59
3.7.3.2 Preparation of Buffers 61
3.7.3.3 Frequently Used Buffers and Solutions 61
3.7.3.4 Calculation of Enzyme Activities 62
3.7.4 Oxidoreductase Assays 64
3.7.4.1 Optical Assays 64
3.7.4.2 Fluorimetric Assays 65
3.7.4.3 Alcohol Dehydrogenase 66
3.7.4.4 Shikimate Dehydrogenase 68
3.7.4.5 L Lactate Dehydrogenase 68
3.7.4.6 Isocitrate Dehydrogenase 71
3.7.4.7 Glucose 6 phosphate Dehydrogenase 72
3.7.4.8 Malate Dehydrogenase 72
3.7.4.9 Glucose Oxidase 73
3.7.4.10 Glyceraldehyde 3 phosphate Dehydrogenase 74
3.7.4.11 Pyruvate Ferredoxin Oxidoreductase 76
3.7.4.12 Glutamate Dehydrogenase 76
3.7.4.13 L Amino Acid Oxidase 77
3.7.4.14 Uricase 78
3.7.4.15 Catalase 79
3.7.4.16 Peroxidase 80
3.7.4.17 Luciferase 82
3.7.5 Pyruvate Dehydrogenase Complex (PDHC) 83
3.7.5.1 Overall Activity of PDHC by NAD+ Reduction 84
3.7.5.2 Overall Activity of PDHC by Dismutation Assay 85
Contents I IX
3.7.5.3 Pyruvate Dehydrogenase (Lipoamide) 86
3.7.5.4 Dihydrolipoamide Acetyltransferase 88
3.7.5.5 Dihydrolipoamide Dehydrogenase 90
3.7.6 a Oxoglutarate Dehydrogenase Complex (OGDHC) 91
3.7.6.1 Overall Activity by NAD+ Reduction 92
3.7.6.2 a Oxoglutarate Dehydrogenase 93
3.7.7 Transferases 94
3.7.7.1 Fatty Acid Synthase 94
3.7.7.2 Phosphorylase a 95
3.7.7.3 Hexokinase 96
3.7.7.4 Pyruvate Kinase 97
3.7.7.5 Acetate Kinase 98
3.7.7.6 3 Phosphoglycerate Kinase 99
3.7.8 Hydrolases 100
3.7.8.1 Lipase 100
3.7.8.2 Cholinesterase 102
3.7.8.3 Acetylcholinesterase 104
3.7.8.4 Alkaline Phosphatase 105
3.7.8.5 Acid Phosphatase 106
3.7.8.6 Ribonudease (Pancreatic) 107
3.7.8.7 a Amylase 108
3.7.8.8 Amyloglucosidase 109
3.7.8.9 Lysozyme 109
3.7.8.10 a Glucosidase 110
3.7.8.11 P Galactosidase 112
3.7.8.12 P Fructosidase 113
3.7.9 Proteases 233
3.7.9.1 Anson Assay 114
3.7.9.2 Casein Assay 116
3.7.9.3 Azocasein Assay 117
3.7.9.4 Ninhydrin Assay 117
3.7.9.5 Leucine Aminopepridase 119
3.7.9.6 Chymotrypsin 129
3.7.9.7 Pepsin 121
3.7.9.8 Trypsin 122
3.7.9.9 Asparaginase 122
3.7.9.10 Urease 123
3.7.9.11 Adenosine Triphosphatase 125
3.7.10 Lyases 126
3.7.10.1 Pyruvate decarboxylase 226
3.7.10.2 Aldolase 127
3.7.10.3 Anthranilate Synthase 127
3.7.10.4 Carbonic Anhydrase 128
3.7.10.5 Fumarase 229
3.7.11 Isomerases 130
xl Contents
3.7.11.1 Glucose/Xylose isomerase 130
3.7.12 Determination of Nicotinamide Nudeotides by Enzymatic Cycling 133
3.7.12.1 Determination of NADP(H) 133
3.7.12.2 Determination of NAD(H) 135
3.8 Diverse Methods 136
3.8.1 Protein Determination 136
3.8.1.1 Biuret Method 136
3.8.1.2 BCA Assay 137
3.8.1.3 Lowry Assay 139
3.8.1.4 Coomassie Binding Assay (Bradford Assay) 340
3.8.1.5 Absorption Method 141
3.8.1.6 Fluorimetric Assay 145
3.8.1.7 Ninhydrin Assay 146
3.8.1.8 Modified Ninhydrin Assay for Protein Determination without
Hydrolysis 147
3.8.1.9 Protein Determination with 2 Hydroxy l naphthaldehyde 148
3.8.2 Phosphate Determination 149
3.8.3 Concentration of Enzyme Solutions 149
3.8.3.1 Precipitation 150
3.8.3.2 Ultrafiltration and Dialysis 153
3.8.3.3 Ultracentrifugation 154
3.8.3.4 Lyophilization 154
3.8.3.5 Miscellaneous Techniques 155
3.9 Enzyme Immunoassays 156
3.9.1 Non competitive Solid phase Enzyme Immunoassay 156
3.9.2 Competitive, Solid phase Enzyme Immunoassay 157
3.9.3 Methods for Enzyme Immunoassays and Immobilization
Techniques 157
3.9.3.1 Protein Coupling to Cyanogen Bromide activated Agarose 157
3.9.3.2 Coupling of Diaminohexyl Spacer 159
3.9.3.3 Periodate Activation of Cellulose 159
3.9.3.4 Conjugation of Proteins (Antibodies) to Enzymes (Peroxidase) 160
3.9.3.5 Introduction of Thiol Groups into Antibodies or Proteins 161
3.9.3.6 Conjugation of |3 galactosidase to Antibodies by MBS 161
3.9.3.7 Conjugation of Alkaline Phosphatase to Antibodies by
Glutaraldehyde 162
3.9.3.8 Cross linking of Proteins with Dimethyl Suberimidate 162
4 Binding Measurements 165
4.1 Irreversible, Reversible, Specific, and Unspecific Binding 365
4.1.1 General Considerations 165
4.1.2 Experimental Aspects 167
4.1.2.1 Amount of Enzyme or Macromolecule 168
4.2.1.2 Stability of the Enzyme or Macromolecule 168
4.1.2.3 Stability of the Ligand 168
Contents I XI
4.1.2.4 Concentration of Components 169
4.2 Binding Measurement by Size Discrimination 170
4.2.1 Ultrafiltration 170
4.2.2 Equilibrium Dialysis 271
4.2.2.1 Determination of Indole Binding to Bovine Serum Albumin 172
4.2.3 Evaluation of Binding Experiments 175
4.2.4 Gel Filtration 177
4.2.5 Ultracentrifugation 179
4.3 Spectroscopic Techniques 180
4.3.1 Difference Spectroscopy 181
4.3.1.1 Difference Spectroscopic Determination of Binding of Ligands to
Catalase 183
4.3.1.2 Evaluation of Spectroscopic Binding Curves 188
4.3.2 Fluorescence Spectroscopy 189
4.3.2.1 ANS Binding to Bovine Serum Albumin 191
4.4 Other Methods for Binding Measurements 195
4.4.1 Radioactive Labeling 195
4.4.2 Reflectrometric Interference Spectroscopy 196
5 Enzymes in Technical Applications 197
5.1 Principles of Enzyme Immobilization 198
5.1.1 Adsorption 199
5.1.2 Entrapment 199
5.1.3 Encapsulation 200
5.1.4 Cross linking 201
5.1.5 Covalent Binding to Solid Supports 201
5.1.5.1 Supports 201
5.1.5.2 Spacer 203
5.2 Methods for Enzyme Immobilization 204
5.2.1 Microencapsulation inside Nylon Beads 205
5.2.2 Entrapment in Polyacrylamide 205
5.2.3 Covalent Immobilization of Enzymes on Nonporous Glass Surfaces 207
5.2.4 Immobilization on Controlled pore Glass 209
5.2.5 Covalent Immobilization of Enzymes to Polyamide 211
5.2.6 O Alkylation with Triethyloxoniumtetrafluoroborate 212
5.2.7 Immobilization of Enzymes to Amine Groups after Partial Hydrolysis of
Polyamide 216
5.2.8 Immobilization of Enzymes to Carboxyl Groups after Partial Hydrolysis
of Polyamide 217
5.2.9 Immobilization of Enzymes to Polyester 218
5.2.10 Immobilization by Alkaline Hydrolysis and Activation with
Tosylchloride 220
5.2.11 Alkaline Hydrolysis and Activation by Carbonyldiimidazol 221
5.3 Analysis of Immobilized Enzymes 222
5.3.1 Protein Assays 222
XIII Contents
5.3.2 Enzyme Assays 222
5.3.2.1 Modified Optical Assay for Immobilized Enzymes 223
5.3.2.2 Cofactors in Reactions with Immobilized Enzymes 224
5.4 Enzyme Reactors 225
5.4.1 Batch Reactor (Stirred tank Reactor) 226
5.4.2 Membrane Reactor 226
5.4.3 Solid bed Reactor 227
5.4.4 Immobilized Cells 227
5.5 Biosensors 228
5.5.1 Enzyme Electrodes 228
5.5.2 Immunoelectrodes 232
5.5.3 Other Biosensors 233
5.5.4 Bioaffinity Sensors 233
5.6 Immobilized Enzymes in Therapy 234
Index 249
|
| any_adam_object | 1 |
| author | Bisswanger, Hans 1943- |
| author_GND | (DE-588)129060461 |
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| callnumber-subject | QP - Physiology |
| classification_rvk | VK 8700 WC 4350 WD 5050 |
| classification_tum | CHE 828f |
| ctrlnum | (OCoLC)249600400 (DE-599)BVBBV017371359 |
| dewey-full | 572.7 |
| dewey-hundreds | 500 - Natural sciences and mathematics |
| dewey-ones | 572 - Biochemistry |
| dewey-raw | 572.7 |
| dewey-search | 572.7 |
| dewey-sort | 3572.7 |
| dewey-tens | 570 - Biology |
| discipline | Chemie / Pharmazie Biologie Chemie |
| format | Book |
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| language | English |
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| publisher | Wiley-VCH |
| record_format | marc |
| spelling | Bisswanger, Hans 1943- Verfasser (DE-588)129060461 aut Practical enzymology Hans Bisswanger Weinheim Wiley-VCH 2004 XV, 255 S. graph. Darst. txt rdacontent n rdamedia nc rdacarrier Enzym Enzymtechnologie (DE-588)4135962-8 gnd rswk-swf Enzym (DE-588)4014988-2 gnd rswk-swf Enzymkinetik (DE-588)4133166-7 gnd rswk-swf 1\p (DE-588)4127380-1 Praktikum gnd-content 2\p (DE-588)4123623-3 Lehrbuch gnd-content Enzym (DE-588)4014988-2 s DE-604 Enzymtechnologie (DE-588)4135962-8 s Enzymkinetik (DE-588)4133166-7 s 3\p DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010469882&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis 1\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 2\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk 3\p cgwrk 20201028 DE-101 https://d-nb.info/provenance/plan#cgwrk |
| spellingShingle | Bisswanger, Hans 1943- Practical enzymology Enzym Enzymtechnologie (DE-588)4135962-8 gnd Enzym (DE-588)4014988-2 gnd Enzymkinetik (DE-588)4133166-7 gnd |
| subject_GND | (DE-588)4135962-8 (DE-588)4014988-2 (DE-588)4133166-7 (DE-588)4127380-1 (DE-588)4123623-3 |
| title | Practical enzymology |
| title_auth | Practical enzymology |
| title_exact_search | Practical enzymology |
| title_full | Practical enzymology Hans Bisswanger |
| title_fullStr | Practical enzymology Hans Bisswanger |
| title_full_unstemmed | Practical enzymology Hans Bisswanger |
| title_short | Practical enzymology |
| title_sort | practical enzymology |
| topic | Enzym Enzymtechnologie (DE-588)4135962-8 gnd Enzym (DE-588)4014988-2 gnd Enzymkinetik (DE-588)4133166-7 gnd |
| topic_facet | Enzym Enzymtechnologie Enzymkinetik Praktikum Lehrbuch |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=010469882&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT bisswangerhans practicalenzymology |