Contents
Preface
Chapter1 Introduction 1
1.1 Conception of Enzyme and Its Development 2
1.2 The Characteristics of Enzyme Action 3
1.3 Factors that Affecting Enzyme Action 7
1.4 Classification and Nomenc1ature of Enzymes 11
1.5 Determination of Enzyme Activity 19
1.6 Methods for Enzymes Production 24
1.7 Development Survey of Enzyme Engineering 28
Review and Thinking 32
Chapter 2 Enzyme Production by Microbial Fermentation33
2.1 Regulation of Enzyme Biosynthesis in Microbial Cells 34
2.2 Characteristic of Microbes for Enzyme Production 49
2.3 Fermentation Process Conditions and Its Connol 53
2.4 Enzyme Fermentation Kinetics 65
2.5 Enzyme Production by Immobilized Cell Fermentation 71
2.6 Enzyme Production by Innnobilized Protoplast Fermentation 78
Review and Thinking 80
Chapter 3 Enzyme Production by Animal and Plant CelI Cultures 81
3.1 The Regulation of Enzyme Biosynthesis in Animal and Plant Cells 82
3.2 Enzyme Production by Plant Cell Culture 86
3.3 Enzyme Production by Animal Cell Culture 97
Review and Thinking ..105
Chapter 4 Extraction, Separation and purification，tioo of Enzymes 106
4.1 Cell Disruption 106
4.2 Enzyme Extraction 110
4.3 Precipitation Separation 113
4.4 Centrifugal Separation 118
4.5 Filtration and Menlbrane Separation 124
4.6 Chrornatographic Separation 130
4.7 Electrophoresis Separation 150
4.8 Exnaction Separation 160
4.9 Crystallization 167
4.10 Concentration and Drying 170
Review and Thinking 172
Chapter 5 Enzyme Molecular Modification 174
5.1 Metal Ion Substitution Modification 175
5.2 Macromolecular Binding Modification 177
5.3 Side Chain Group Modification 181
5.4 Peptide Chain Limited Hydrolysis Modification 187
5.5 Nucleotide Chain Cleavage Modification 188
5.6 Amino Acid Substitution Modification 189
5.7 Nucleotide Substitution Modification 192
5.8 Physical Modification 193
5.9 Application of Enzyme Molecular Modification 194
Review and Thlnking 200
Chapter 6 ImmobilizatiOll of Enzymes, cells and Protoplasts 201
6.1 Enzyme Immobilization 203
6.2 Cell Immo biliza tion 216
6.3 Protoplast Immobilization 228
Review and Thlnking 231
Chapter 7 Enzyme Catalysis in Non-aqueous phases 232
7.1 The Main Scopes of Enzyme Catalysis in Non-aqueous Phase 233
7.2 Effects of Water and Organic Solvents on Enzyme Catalysis in Organic Media 234
7.3 The Catalytic Characters of Enzymes in Organic Media 240
7.4 Conditions of Enzyme Catalytic Reaction in Organic Media and Its Control 244
7.5 Application of Enzyme Catalysis in Non-aqueous Phase 251
Review and Thlnking 260
Chapter 8 Enzyme Directed Evolotion 261
8.1 The characteristics of Enzyme Directed Evolution 263
8.2 Random Mutation of Enzyme Gene 264
8.3 Directed Selection of Enzyme Gene Mutants 270
8.4 The Applications of Enzyme Directed Evolution 281
Review and Thlnking 283
Chapter 9 Enzyme Reactors 285
9.1 Types of Enzyme Reactors 286
9.2 Choice of Enzyme Reactors 292
9.3 Design of Enzyme Reactors 296
9.4 Enzyme Reactor Operation 301
Review and Thlnking 306
Chapter 10 Application of Enzymes 307
10.1 Enzymes Application in Pharmaceutical Industry 307
10.2 Application of Enzymes in Food 328
10.3 Enzyme Application in Light and Chemical Industries 342
10.4 Enzyme Application in Environmental Protection 349
10.5 Enzyme Application in Biotechnology 352
Review and Thinking 358
Main references 359