Directed Molecular Evolution of Proteins: Or How to Improve Enzymes for Biocata

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Book Title
Directed Molecular Evolution of Proteins: Or How to Improve Enzym
ISBN
9783527304233
Categoria

Informazioni su questo prodotto

Product Identifiers

Publisher
Wiley & Sons, Incorporated, John
ISBN-10
3527304231
ISBN-13
9783527304233
eBay Product ID (ePID)
2181052

Product Key Features

Number of Pages
368 Pages
Publication Name
Directed Molecular Evolution of Proteins : or How to Improve Enzymes for Biocatalysis
Language
English
Publication Year
2002
Subject
Chemistry / Organic
Type
Textbook
Subject Area
Science
Author
Kai Johnsson
Format
Hardcover

Dimensions

Item Height
0.9 in
Item Weight
29.8 Oz
Item Length
9.6 in
Item Width
7 in

Additional Product Features

Intended Audience
Scholarly & Professional
LCCN
2002-280625
Dewey Edition
21
Illustrated
Yes
Dewey Decimal
547.750442
Table Of Content
List of Contributors xi 1 Introduction 1 2 Evolutionary Biotechnology - From Ideas and Concepts to Experiments and Computer Simulations 5 2.1 Evolution in vivo - From Natural Selection to Population Genetics 5 2.2 Evolution in vitro - From Kinetic Equations to Magic Molecules 8 2.3 Evolution in silico - From Neutral Networks to Multi-stable Molecules 16 2.4 Sequence Structure Mappings of Proteins 25 2.5 Concluding Remarks 26 3 Using Evolutionary Strategies to Investigate the Structure and Function of Chorismate Mutases 29 3.1 Introduction 29 3.2 Selection versus Screening 30 3.3 Genetic Selection of Novel Chorismate Mutases 33 3.4 Summary and General Perspectives 57 4 Construction of Environmental Libraries for Functional Screening of Enzyme Activity 63 4.1 Sample Collection and DNA Isolation from Environmental Samples 65 4.2 Construction of Environmental Libraries 68 4.3 Screening of Environmental Libraries 71 4.4 Conclusions 76 5 Investigation of Phage Display for the Directed Evolution of Enzymes 79 5.1 Introduction 79 5.2 The Phage Display 79 5.3 Phage Display of Enzymes 81 5.4 Creating Libraries of Mutants 87 5.5 Selection of Phage-enzymes 89 5.6 Conclusions 108 6 Directed Evolution of Binding Proteins by Cell Surface Display: Analysis of the Screening Process 111 6.1 Introduction 111 6.2 Library Construction 113 6.3 Mutant Isolation 115 6.4 Summary 124 7 Yeast n-Hybrid Systems for Molecular Evolution 127 7.1 Introduction 127 7.2 Technical Considerations 130 7.3 Applications 147 7.4 Conclusion 155 8 Advanced Screening Strategies for Biocatalyst Discovery 159 8.1 Introduction 159 8.2 Semi-quantitative Screening in Agar-plate Formats 161 8.3 Solution-based Screening in Microplate Formats 164 8.4 Robotics and Automation 169 9 Engineering Protein Evolution 177 9.1 Introduction 177 9.2 Mechanisms of Protein Evolution in Nature 178 9.3 Engineering Genes and Gene Fragments 187 9.4 Gene Fusion ± From Bi- to Multifunctional Enzymes 203 9.5 Perspectives 208 10 Exploring the Diversity of Heme Enzymes through Directed Evolution 215 10.1 Introduction 215 10.2 Heme Proteins 216 10.3 Cytochromes P450 218 10.4 Peroxidases 223 10.5 Comparison of P450s and Peroxidases 227 10.6 Chloroperoxidase 228 10.7 Mutagenesis Studies 229 10.8 Directed Evolution of Heme Enzymes 233 10.9 Conclusions 238 11 Directed Evolution as a Means to Create Enantioselective Enzymes for Use in Organic Chemistry 245 11.1 Introduction 245 11.2 Mutagenesis Methods 247 11.3 Overexpression of Genes and Secretion of Enzymes 248 11.4 High-Throughput Screening Systems for Enantioselectivity 250 11.5 Examples of Directed Evolution of Enantioselective Enzymes 257 11.6 Conclusions 273 12 Applied Molecular Evolution of Enzymes Involved in Synthesis and Repair of DNA 281 12.1 Introduction 281 12.3 Directed Evolution of DNA polymerases 289 12.4 Directed Evolution of Thymidine Kinase 295 12.5 Directed Evolution of Thymidylate Synthase 297 12.6 O 6 -Alkylguanine-DNA Alkyltransferase 300 12.7 Discussion 302 13 Evolutionary Generation versus Rational Design of Restriction Endonucleases with Novel Specificity 309 13.1 Introduction 309 13.2 Design of Restriction Endonucleases with New Specificities 313 13.3 Summary and Outlook 324 14 Evolutionary Generation of Enzymes with Novel Substrate Specificities 329 14.1 Introduction 329 14.2 General Considerations 331 14.3 Examples 333 14.4 Conclusions 339 Index 343
Synopsis
Natural selection created optimal catalysts. However, optimal performance of enzyme catalysis does not necessarily refer to maximum reaction rate. Rather, it may be a compromise between specificity, rate, stability, and other chemical constraints that makes enzymes capable of catalyzing reactions under mild conditions and with high substrate specificity, accompanied by high regio- and enantioselectivity. The book presented here focuses on the directed evolution of proteins, which has established itself as a powerful method for designing enzymes showing new substrate specificities. It includes a comprehensive repertoire of techniques for producing combinatorial enzyme libraries, while the functional gene expression in a suitable host helps in selecting the appropriate structure, making fast screening a necessity. This book illustrates both the theoretical background as well as the potential of this interesting method in practice - which is becoming ever more important even in classical organic synthesis, Die natürliche Auslese hat optimale Protein-Katalysatoren hervorgebracht; eine optimale Enzymkatalyse indes verläuft nicht notwendigerweise mit der höchstmöglichen Geschwindigkeit. Vielmehr ist es oft ein Kompromi zwischen Spezifität, Geschwindigkeit, Stabilität und vielen anderen Faktoren, den die Natur für eine chemische Reaktion unter milden Bedingungen, mit hoher Regio- und Enantioselektivität, eingeht. Vor diesem Hintergrund hat sich die gerichtete Evolution von Proteinen als eine leistungsfähige Methode für das Design von Enzymen mit neuen Substratspezifitäten entwickelt. Das vorliegende Buch beleuchtet das umfassende Repertoire an Techniken für das Erstellen von kombinatorischen Enzymbibliotheken, angefangen von den theoretischen Grundlagen bis hin zur praktischen Anwendung - die immer mehr auch in der klassischen organischen Synthese an Bedeutung gewinnt!, Natural selection created optimal catalysts. However, optimal performance of enzyme catalysis does not necessarily refer to maximum reaction rate. Rather, it may be a compromise between specificity, rate, stability, and other chemical constraints that makes enzymes capable of catalyzing reactions under mild conditions and with high substrate specificity, accompanied by high regio- and enantioselectivity. The book presented here focuses on the directed evolution of proteins, which has established itself as a powerful method for designing enzymes showing new substrate specificities. It includes a comprehensive repertoire of techniques for producing combinatorial enzyme libraries, while the functional gene expression in a suitable host helps in selecting the appropriate structure, making fast screening a necessity. This book illustrates both the theoretical background as well as the potential of this interesting method in practice - which is becoming ever more important even in classical organic synthesis!, Natural selection created optimal catalysts. However, optimal performance of enzyme catalysis does not necessarily refer to maximum reaction rate.
LC Classification Number
TP248.65.E59D545

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