Practical Biotransformations -- Contents -- Foreword -- Preface -- 1 Biotransformations, Microbes and Enzymes -- 1.1 Introduction -- 1.2 Biotransformations -- 1.3 Microorganisms -- 1.4 Organism Nomenclature -- 1.5 Enzymes -- 1.6 Types of Enzymatic Reactions -- 1.7 Enzymatic Cofactors -- 1.8 Some Basic Characteristics of Enzyme Catalysis -- 1.9 Types of Biocatalyst - Biotransformations by 'Whole Cells' or 'Isolated Enzymes' -- 1.10 Conclusion -- References -- 2 An Overview of Biocatalyst Sources and Web-Based Information -- 2.1 Introduction -- 2.2 Microbial Culture Collections -- 2.3 Obtaining Organisms from Other Research Groups -- 2.4 Selective Enrichments -- 2.5 Metagenomics -- 2.6 Enzyme Suppliers and Biocatalyst Development Companies -- 2.7 Genome Mining for Biocatalysts -- 2.8 Obtaining Amino Acid and Gene Sequence Information on Biocatalysts -- 2.9 Obtaining DNA Templates for Cloning -- 2.10 Custom Gene Synthesis -- 2.11 Other Web Resources for Finding Information on Biocatalysts -- 2.12 Conclusion -- References -- 3 Setting up a Laboratory for Biotransformations -- 3.1 Introduction -- 3.2 Microbiological Containment -- 3.3 On Containment Issues and Genetically Modified Organisms (GMOs) -- 3.4 Equipment for Handling Microorganisms -- 3.5 Techniques and Terms in Microbiology - Sterility, Asepsis and Aseptic Technique -- 3.6 Disposal of Viable Microbial Waste and Disinfection of Reusable Equipment -- 3.7 Equipment for Enzymology and Molecular Biology -- 3.8 General Reagents and Chemicals in a Biotransformations Laboratory -- 3.9 Conclusion -- References -- 4 A Beginner's Guide to Preparative Whole-Cell Microbial Transformations -- 4.1 Introduction -- 4.2 Storage, Maintenance and Growth of Microorganisms -- 4.3 General Microbiological Methods -- 4.4 Examples of Whole-Cell Biotransformations Using Bacteria.

4.5 Biotransformation by Filamentous Fungi and Yeasts -- 4.6 Whole-Cell Biotransformations by Recombinant Strains of E. Coli -- 4.7 Conclusion -- References -- 5 A Beginner's Guide to Biotransformations Catalysed by Commercially Available Isolated Enzymes -- 5.1 Introduction -- 5.2 Lipases -- 5.3 Hydrolytic Reactions Using Lipases -- 5.4 Using Lipases for Acylation Reactions -- 5.5 Other Hydrolases -- 5.6 Commercially Available Coenzyme-Dependent Enzymes -- 5.7 Carbon-Carbon Bond Forming Reactions -- 5.8 Conclusion -- References -- 6 A Beginner's Guide to the Isolation, Analysis and Use of Home-Grown Enzyme Biocatalysts -- 6.1 Introduction -- 6.2 Cell Growth and Harvesting -- 6.3 Cell Disruption -- 6.4 A Typical Procedure for Making a Cell Extract from a Recombinant Strain of E. Coli -- 6.5 Puri.cation of Enzymes - A Brief Guide -- 6.6 Techniques for Protein Purification -- 6.7 Isolation of Recombinant Enzymes Using Histidine Tags -- 6.8 Estimation of Protein Concentration -- 6.9 Concentrating Protein Samples by Centrifugation -- 6.10 Analysis of Protein Samples by Sodium Dodecylsulfate Polyacylamide Gel Electrophoresis -- 6.11 Examples of Enzyme Assays -- 6.12 Using Home-Grown Enzymes for Biotransformations - Some Recent Examples -- 6.13 Conclusion -- References -- 7 An Introduction to Basic Gene Cloning for the Production of Designer Biocatalysts -- 7.1 Introduction -- 7.2 Background -- 7.3 Gene Amplification by Polymerase Chain Reaction (PCR) -- 7.4 DNA Fragment Analysis by Agarose Gel Electrophoresis -- 7.5 Gene Cloning -- 7.6 Analysis by DNA Sequencing -- 7.7 Troubleshooting the Gene Amplification and Cloning Process -- 7.8 Ligation-Independent Cloning -- 7.9 Gene Expression -- 7.10 Conclusion -- References -- 8 Engineering Enzymes -- 8.1 Introduction.

8.2 Site-Directed or Targeted Mutagenesis as a Tool for Investigating Enzyme Mechanism or Altering Catalytic Attributes -- 8.3 Engineering Using Random Mutagenesis Directed Evolution of Enzymes -- 8.4 Combining Rational and Random Mutagenesis for Biocatalyst Improvement -- 8.5 Exploiting Catalytic Promiscuity for Creating New Enzyme Activities -- 8.6 Designing Enzymes in Silico -- 8.7 Conclusion -- References -- Appendices -- Index.