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