Principles of Biotechnology

The first edition of this book appeared in 1983, and provided the first easily-accessible account of the state of biotechnology at a level suitable for advanced undergraduates and postgraduates. In this new edition, specialists in biotechnology at the University of Surrey have again collaborated with industrial experts to provide authoritative interdisciplinary coverage of an ever-expanding field. The revision of the text reflects the rapid advances made in many of the fields which go to make up biotechnology-ranging from the molecular to the process plant level. Biotechnology is an applied science, and this book relates theory to application by placing the basic advances in the context of commercial applicability and process optimization. For those wishing to explore the literature further, references to up-to-date reviews and original research publications are provided. Special thanks are due not only to the contributors, but to all those who have helped in the planning of this book.AW v Background to authors C. Bucke is Principal Lecturer in the School of Biotechnology, Polytechnic of Central London, and was previously Programme Manager Biotechnology at Tate & Lyle Group Research and Development. His research interests include the production and use of immobilized biocatalysts, the use of enzymes in the synthesis of novel bisaccharides and polysaccharides and the use of enzymes in extreme environments. He is Scientific Co-ordinator of the Institute for Biotechnological Studies, Department of Trade and Industry 'Extended use of Biocatalysts' Programme.

1 Features of biotechnology and its scientific basis.- 1.1 Introduction.- 1.2 Interrelationships between microorganisms and enzymes.- 1.3 Success in biotechnology.- 2 Application of the principles of industrial microbiology to biotechnology.- 2.1 Introduction.- 2.2 Primary metabolism.- 2.2.1 Introduction.- 2.2.2 The batch culture.- 2.3 Secondary metabolism.- 2.3.1 Products.- 2.3.2 Definitions of a secondary metabolite.- 2.3.3 Regulatory factors.- 2.4 Primary metabolites in industrial biotechnology.- 2.4.1 Potable alcohol.- 2.4.2 Amino acids.- 2.4.3 Other organic acids.- 2.4.4 Polysaccharides.- 2.4.5 Other primary metabolites.- 2.4.6 Single cell protein (SCP).- 2.4.7 Future plans in the UK.- 2.5 Secondary metabolites in biotechnology.- 2.5.1 Penicillin.- 2.5.2 Other secondary metabolites.- 3 Application of the principles of microbial genetics to biotechnology.- 3.1 Control mechanisms in microorganisms.- 3.2 Manipulations in vivo.- 3.2.1 Mutation.- 3.2.2 Recombination.- 3.2.3 Application of manipulations in vivo to increase enzyme production.- 3.2.4 Overproduction of primary metabolites.- 3.2.5 Overproduction of secondary metabolites.- 3.2.6 Production of novel metabolites.- 3.3 Manipulations in vitro.- 3.3.1 Basic techniques of in-vitro genetic manipulation.- 3.3.2 Uses of DNA cloning.- 3.3.3 Genes and biotechnology: applications of gene cloning.- 3.3.4 Safety implications.- 3.3.5 Future prospects.- 4 Application of the principles of fermentation engineering to biotechnology.- 4.1 The fermenter.- 4.1.1 The environment.- 4.1.2 Principal types of fermenter.- 4.1.3 Mode of operation.- 4.1.4 Ancillary processes.- 4.2 General design principles.- 4.2.1 Basic design rules.- 4.2.2 Materials and components.- 4.2.3 Control and instrumentation.- 4.2.4 Stirred-tank fermenters.- 4.2.5 Gas-lift and sparged-tank fermenters.- 4.3 Heat transfer.- 4.3.1 Heat sterilization.- 4.3.2 Sterilization without heat.- 4.4 Mixing.- 4.4.1 Introduction.- 4.4.2 Outline of rheology.- 4.5 Oxygen supply in fermenters.- 4.6 Scale-up in biotechnology.- 4.6.1 The problems of large-scale operation.- 4.6.2 Selection of scale-up criteria.- 4.6.3 Interaction of criteria.- 4.7 Fermentation processes in biotechnology.- 4.7.1 Brewing.- 4.7.2 Penicillin manufacture.- 4.7.3 Biological waste-water treatment.- 4.7.4 Single-cell protein (SCP) production.- 4.7.5 Outlook for biotechnology.- 4.8 Summary.- 5 Application of the principles of enzymology to biotechnology.- 5.1 Features of enzymes in relation to biotechnology.- 5.1.1 Introduction.- 5.1.2 Advantages of using enzymes for manufacture of products.- 5.1.3 Choice and control of enzymes in applications.- 5.2 Applications of enzymes in biotechnology.- 5.2.1 Large-scale industrial applications.- 6 The biotechnology of enzyme isolation and purification.- 6.1 Introduction.- 6.2 Enzyme sources.- 6.3 Release of enzymes from cells.- 6.3.1 Sources.- 6.3.2 Extraction by physical methods.- 6.3.3 Extraction by chemical methods.- 6.4 Primary clarification of the soluble enzyme.- 6.4.1 Centrifugation.- 6.4.2 Flocculation and coagulation.- 6.4.3 Filtration.- 6.5 Concentration.- 6.5.1 Removal of nucleic acids.- 6.5.2 Precipitation.- 6.5.3 Ultrafiltration and reverse osmosis.- 6.5.4 Freeze-drying.- 6.5.5 Evaporation.- 6.5.6 Freezing.- 6.6 Enzyme purification—chromatography.- 6.6.1 Gel chromatography.- 6.6.2 Ion exchange chromatography.- 6.6.3 Affinity purification.- 6.6.4 Chromatography columns.- 6.6.5 High-performance liquid chromatography of enzymes.- 7 The application of immobilized enzymes, immobilized cells and biochemical reactors in biotechnology—principles of enzyme engineering.- 7.1 Introduction.- 7.2 The application of biological catalysts.- 7.3 Types of enzymic catalyst and commercial applications.- 7.3.1 Immobilized biocatalysts.- 7.3.2 Assessment of supports and methods.- 7.3.3 Effectiveness factors for immobilized enzymes.- 7.3.4 The kinetics of enzymes in industrial use.- 7.3.5 Factors which modify the intrinsic activity of enzymes.- 7.3.6 The stability of immobilized biocatalyst—diffusion artifacts.- 7.4 Enzyme reactors.- 7.4.1 Types of biochemical reactor.- 7.4.2 Assessment of the performance of biochemical reactors.- 7.4.3 Practical enzyme reactor kinetics.- 7.4.4 The effect of non-ideal flow on biochemical reactor performance.- 7.4.5 The stability of biochemical reactors.- 7.4.6 Physical problems associated with the use of immobilized biocatalysts in biochemical reactors.- 7.4.7 Purification and recovery of the products of biochemical reactors (downstream processing).- 7.5 Conclusions.