Practical Biotransformations A Beginner's Guide

Name: Practical Biotransformations A Beginner's Guide
Price: 128.77 USD
Availability: InStock
ISBN: 9781405193672

ISBN-10: 1405193670

ISBN-13: 9781405193672

Edition: 2009 (Guide (Instructor's))

Authors: Gideon Grogan

List price: $265.95

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Description:

Biotransformations are reactions catalysed by either enzymes or microbes which are of potential use to preparative synthetic chemistry. Biotransformation science can include microbiological and biochemical techniques with which the organic chemist is unfamiliar, and can result in significant research and economic benefits. The book assists chemistry postgraduates and researchers in both academia and industry who understand biotransformation methodology and its value, but do not know how to implement the relevant experimental techniques in the laboratory. It is written in an accessible and user-friendly way that should facilitate the introduction of biocatalysis into labs.

Book details

List price: $265.95
Copyright year: 2009
Publisher: John Wiley & Sons, Incorporated
Publication date: 3/31/2009
Binding: Hardcover
Pages: 344
Size: 6.20" wide x 9.20" long x 0.90" tall
Weight: 1.408
Language: English

Gideon Grogan is a lecturer at the University of York, Department of Chemistry. He is also Science Manager of the Bioscience for Business Knowledge Transfer Network (a UK DTI-funded organisation for biocatalysis collaborations between academe and industry) and Deputy Director of the Centre of Excellence for Biocatalysis, Biotransformation and Biomanufacture (at York).




Biotransformations, Microbes and Enzymes



Introduction



Biotransformations



Microorganisms



Organism nomenclature



Enzymes



Types of Enzymatic reactions



Enzymatic Cofactors



Some Basic Characteristics of Enzyme Catalysis



Types of Biocatalyst - Biotransformations by 'whole cells' or isolated enzymes



Conclusion



An overview of biocatalyst sources and web-based information



Introduction



Microbial culture collections



Obtaining organisms from other research groups



Selective Enrichments



Metagenomics



Enzyme Suppliers and Biocatalyst Development Companies



Genome mining for biocatalysts



Obtaining amino acid and gene sequence information on biocatalysts



Obtaining DNA templates for cloning



Custom Gene Synthesis



Other interesting web resources for biocatalysis



Conclusion



Setting up a laboratory for biotransformations



Introduction



Microbiological Containment



On containment issues and genetically-modified organisms



Equipment for handling microorganisms



Techniques and terms in microbiology - Sterility, Asepsis and Aseptic Technique



Disposal of viable microbial waste and disinfection of reusable equipment



Equipment for enzymology and molecular biology



General reagents and chemicals in a Biotransformations Laboratory



Conclusion



A beginner's guide to preparative whole-cell microbial biotransformations



Introduction



Storage, maintenance and growth of microorganisms



General Microbiological Methods



Examples of Whole-cell Biotransformations using Bacteria



Biotransformation by filamentous fungi and yeasts



Whole-cell Biotransformations by recombinant strains of E. coli



Conclusion



A beginner's guide to biotransformations by commercially available isolated enzymes



Introduction



Lipases



Hydrolytic Reactions using lipases



Using lipases for acylation reactions



Other hydrolases



Commercially available Coenzyme-dependent Enzymes



Carbon-carbon bond forming reactions



Conclusion



A beginner's guide to the isolation and analysis and use of home-grown enzyme biocatalysts



Introduction



Cell growth and harvesting



Cell disruption



A typical procedure for making a cell extract from a recombinant strain of E. coli



Purification of enzymes - a brief guide



Techniques for Protein Purification



Isolation of recombinant enzymes using histidine tags



Estimation of protein concentration



Concentrating protein samples by centrifugation



Analysis of protein samples by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE)



Examples of enzyme assays



Using home-grown enzymes for biotransformations ? Some recent examples



Conclusion



An introduction to basic gene cloning for the production of designer biocatalysts



Introduction



Background to gene cloning



Gene amplification by polymerase chain reaction (PCR)



DNA fragment analysis by agarose electrophoresis



Gene cloning



Analysis by DNA sequencing



Troubleshooting the gene amplification and cloning process



Ligation-Independent Cloning



Gene Expression in E. coli



Conclusion



Engineering Enzymes



Introduction



Site-directed or targted mutagenesis as a tool for investigating enzyme mechanism or altering catalytic attributes



A site-directed mutagenesis experiment. Considerations and practise



Engineering using random mutagenesis. Directed Evolution of Enzymes



Combining rational and random mutagenesis for biocatalyst improvement



Exploiting catalytic promiscuity for creating new enzyme activities



Designing enzymes in silico



Conclusion


Appendices



Structures of the proteinogenic amino acids



Structures of bases found in nucleic acids



The Genetic Code



Recipes for Microbiological Growth Media



Biological buffers



Ammonium sulphate fractionation table



Restriction enzymes and restriction sites