Part 1 Fundamentals and principles 1
1 Public perception of biotechnology JOHN E．SMITH 3
1.1 Introduction 3
1.2 Public awareness of genetic engineering 4
1.3 Regulatory requirements-safety of genetically engineered foods 7
List of contributors 9
Contents 9
1.4 Labelling-how far should it go? 10
1.5 Policy making 11
1.6 Areas of significant public concern 12
Preface 13
1.7 Conclusions 16
1.8 Further reading 16
2.1 Introduction 17
2 Biochemistry and physiology of growth and metabolism COLIN RATLEDGE 17
2.2 Metabolism 18
2.3 Catabolic pathways 21
2.4 Gluconeogenesis 29
2.5 Energy production in aerobic micro-organisms 29
2.6 Anaerobic metabolism 32
2.7 Biosynthesis 35
2.8 Control of metabolic processes 38
2.9 Efficiency of microbial growth 43
2.10 Further reading 44
3 Stoichiometry and kinetics of microbial growth from a thermodynamic perspective J．J．HEIJNEN 45
Nomenclature 45
3.1 Introduction 45
3.2 Stoichiometry calculations 46
3.3 Stoichiometry predictions based on Gibbs energy dissipation 52
3.4 Growth kinetics from a thermodynamic point of view 56
3.5 Further reading 57
4 Genome management and analysis:prokaryotes COLIN R．HARWOOD and ANIL WIPAT 59
4.1 Introduction 59
4.2 Bacterial chromosomes and natural gene transfer 59
4.3 What is genetic engineering and what is it used for? 65
4.4 The basic tools of genetic engineering 66
4.5 Cloning vectors and libraries 76
4.6 Analysis of genomes/proteomes 81
4.7 Analysis of gene expression 83
4.8 Engineering genes and optimising products 87
4.9 Production of heterologous products 89
4.10 In silico analysis of bacterial genomes 92
4.11 Further reading 93
Glossary 95
5 Genetic engineering:yeasts and filamentous fungi DAVID B．ARCHER DONALD A．MACKENZIE and DAVID J．JEENES 95
5.1 Introduction 97
5.2 Introducing DNA into fungi(fungal transformation) 100
5.3 Gene cloning 107
5.4 Gene structure,organisation and expression 116
5.5 Special methodologies 119
5.6 Biotechnological applications of fungi 120
5.7 Further reading 126
6 Microbial process kinetics JENS NIELSEN 127
Nomenclature 127
6.1 Introduction 128
6.2 Kinetic modelling of cell growth 130
6.3 Mass balances for ideal bioreactors 141
6.4 Further reading 149
Nomenclature 151
7 Bioreactor design YUSUF CHISTI and MURRAY MOO-YOUNG 151
7.1 Introduction 152
7.2 Bioreactor configurations 153
7.3 Bioreactor design features 158
7.4 Design for sterile operation 159
7.5 Photobioreactors 162
7.6 Heat transfer 164
7.7 Shear effects in culture 168
7.8 Further reading 171
8 Mass transfer HENK J．NOORMAN 173
Nomenclature 173
8.1 Introduction 174
8.2 The mass transfer steps 175
8.3 Mass transfer equations 177
8.4 Determining the volumetric mass transfer coefficients 183
8.5 The effect of scale on mass transfer 185
8.6 Further reading 186
9 Downstream processing in biotechnology RAJNI HATTI-KAUL and BO MATTIASSON 187
9.1 Introduction 187
9.2 Downstream processing:a multistage operation 187
9.3 Solid-liquid separation 188
9.4 Release of intracellular components 193
9.5 Concentration of biological products 195
9.6 Purification by chromatography 204
9.7 Product formulation 207
9.8 Monitoring of downstream processing 209
9.9 Process integration 210
9.10 Further reading 211
10 Measurement and control A L？BBERT and R．SIMUTIS 213
Nomenclature 213
10.1 Introduction 214
10.2 Structure of process models 215
10.3 Kinetic rate expressions 216
10.4 Advanced modelling considerations 222
10.5 Process supervision and control 228
10.6 Open-loop control 230
10.7 Closed-loop control 231
10.8 Conclusion 236
10.9 Further reading 238
11 Process economics BJ？RN KRISTIANSENRAJNI 239
11.1 Introduction 239
11.2 The starting point 240
11.3 Cost estimates 240
11.4 Process design 242
11.5 Design exercise 244
11.7 Operating costs estimates 247
11.6 Capital costs estimates 247
11.8 The costs case-to build or not to build 250
11.9 Further reading 252
Part Ⅱ Practical applications 253
12 The business of biotechnology WILLIAM BAINS and CHRIS EVANS 255
12.1 Introduction 255
12.2 What is biotechnology used for? 255
12.3 Biotechnology companies,their care and nurturing 260
12.4 Investment in biotechnology 268
12.5 Who needs management? 275
12.6 Patents and biotechnology 278
12.7 Conclusion:jumping the fence 279
12.8 Further reading 279
13 Amino acids L．EGGELING,W．PFEFFERLE and H．SAHM 281
13.1 Introduction 281
13.2 Commerical use of amino acids 282
13.3 Production methods and tools 284
13.4 L-Glutamate 285
13.5 L-Lysine 289
13.6 L-Threonine 294
13.7 L-Phenylalanine 296
13.8 L-Tryptophan 298
13.9 L-Aspartate 300
13.10 Outlook 302
13.11 Acknowledgements 302
13.12 Further reading 302
14 Organic acids CHRISTIAN P．KUBICEK 305
14.1 Introduction 305
14.2 Citric acid 306
14.3 Gluconic acid 315
14.4 Lactic acid 317
14.5 Other acids 319
14.6 Further reading 324
15 Microbial polyhydroxyalkanoates,polysaccharides and lipids ALISTAIR J．ANDERSON and JAMES P．WYNN 325
15.1 Introduction 325
15.2 Microbial polyhydroxyalkanoates 325
15.3 Microbial polysaccharides 333
15.4 Microbial lipids 339
15.5 Further reading 348
16 Antibiotics DAVID A．LOWE 349
16.1 Introduction 349
16.2 Biosynthesis 351
16.3 Strain improvement 351
16.5 Analysis 354
16.6 Culture preservation and aseptic propagation 354
16.4 Genetic engineenng 354
16.7 Scale-up 355
16.8 Fermentation 356
16.9 Penicillins 361
16.10 Cephalosporins 367
16.11 New β-lactam technologies 368
16.12 Aminoglycosides 370
16.13 Macrolides 373
16.14 Economics 373
16.15 Good Manufactunng Practices 374
16.16 Further reading 375
17 Baker's yeast SVEN-OLOF ENFORS 377
Nomenclature 377
17.1 Introduction 378
17.2 Medium for baker's yeast production 380
17.3 Aerobic ethanol fornation and consumption 381
17.4 The fed-batch technique used to control ethanol production 384
17.5 Industrial process control 386
17.6 Process outline 387
17.7 Further reading 389
18 Production of enzymes DAVID A．LOWE 391
18.1 Introduction 391
18.2 Enzymes from animal and plant sources 393
18.3 Enzymes from microbial sources 395
18.4 Iarge-scaleproduction 396
18.5 Biochemical fundamentals 398
18.6 Genetic engineering 400
18.7 Recoveryofenzymes 402
18.8 Isolation of soluble enzymes 403
18.9 Enzyme purification 404
18.10 Immobilised enzymes 406
18.11 Legislativeand safetyaspects 407
18.12 Further reading 408
19 Synthesis of chemicals using enzymes 409
THORLEIF ANTHONSEN 409
19.1 Introduction 409
19.2 Hydrolytic enzymes 412
19.3 Chiral building blocks for synthesis 414
19.4 Reductions and oxidations 419
19.5 Use ofenzymes in sugar chemistry 422
19.6 Use ofenzymes to make amino acids and peptides 426
19.7 Further reading 428
20 Recombinant proteins of high value GEORG-B．KRESSE 429
20.1 Applications of high-value proteins 429
20.2 Analytical enzymes 430
20.3 Therapeutic proteins 436
20.4 Regulatory aspects of therapeutic proteins 446
20.5 Outlook to the future ofprotein therapies 446
20.6 Further reading 447
21 Mammalian cell culture N．VRIEZEN,J．P．VAN DIJKEN and L．H？GGSTR？M 449
21.1 Introduction 449
21.2 Mammalian cell lines and their characteristics 450
21.3 Commerical products 452
21.4 Protein glycosylation 453
21.5 Media for the cultivation ofmammalian cells 455
21.6 Metabolism 458
21.7 Large-scale cultivation of mammalian cells 462
21.8 Genetic engineering of mammalian cells 468
21.9 Further reading 470
22.1 Introduction 471
22 Biotransformations JOAQUIM M．S．CABRAL 471
22.2 Biocatalyst selection 473
22.3 Biocatalyst immobilisation and performance 475
22.4 Immobilised enzyme reactors 489
22.5 Biocatalysis in non-conventional media 491
22.6 Concluding remarks 500
22.7 Further reading 501
23 Immunochemical applications MIKE CLARK 503
Glossary 503
23.1 Introduction 506
23.2 Antibody structure and functions 506
23.3 Antibody protein fragments 510
23.4 Antibody affinity 512
23.5 Antibody specificity 513
23.6 Immunisation and production ofpolyclonal antisera 514
23.7 Monoclonal antibodies 517
23.8 Antibody engineering 518
23.9 Combinatorial and phage display libraries 520
23.10 In vitro uses of recombinant and monoclonal antibodies 522
23.11 In vivo uses ofrecombinant and monoclonal antibodies 526
23.12 Further reading 529
24 Environmental applications PHILIPPE VANDEVIVERE and WILLY VERSTRAETE 531
24.1 Introduction 531
24.2 Treatment of waste water 532
24.3 Digestion of organic slurries 540
24.4 Treatment of solid wastes 542
24.5 Treatment of waste gases 545
24.6 Soil remediation 549
24.7 Treatment of groundwater 554
24.8 Further reading 557
Index 559