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Practical Biotransformations A Beginner's Guide

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ISBN-10: 1405193670

ISBN-13: 9781405193672

Edition: 2009 (Guide (Instructor's))

Authors: Gideon Grogan

List price: $255.95
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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.
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Book details

List price: $255.95
Copyright year: 2009
Publisher: John Wiley & Sons, Limited
Publication date: 3/31/2009
Binding: Hardcover
Pages: 344
Size: 6.30" wide x 9.30" long x 1.00" 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