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Directed Evolution of Proteins

Directed Evolution of Proteins

Marco Fioroni, Raluca Ostafe, Radivoje Prodanovic, Ulrich Schwaneberg

ISBN: 978-3-527-32369-2

Jun 2015

400 pages

Select type: Hardcover

$105.00

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Description

Aimed at providing researchers and students in academia and industry with an in-depth understanding of directed protein evolution, this advanced textbook enables the design of promising experiments and advanced technologies in diversity generation and high-throughput screening. It contains benchmarking systems and computational tools for comparing the performance of random mutagenesis methods, a section on focused mutant library design and concludes with a look at the challenges, barriers and promising strategies for directed evolution
Clearly written and richly illustrated, this book also includes many pedagogical features, such as information boxes summarizing the important information in each chapter.
DIRECTED PROTEIN EVOLUTION
History and Landmarks in Directed Protein Evolution
Industrial and Academic Impact of Directed Protein Evolution
General Consideration on whether and how to start a Directed Evolution Experiment
Exercises
DIVERSITY GENERATION
Introduction to Sequence Space Generation and Exploration
Classification of Random Mutagenesis Method
Exercises
BENCHMARKING SYSTEMS AND COMPUTATIONAL TOOLS FOR COMPARING THE PERFORMANCE OF RANDOM MUTAGENESIS METHODS
DNA-Based Benchmarking Systems
Protein-Based Benchmarking Systems
Analysis and Performance of Random Mutagenesis Methods
Strategies for Complementary Diversity Generation
Exercises
Computational Tools for Random Diversity Generation
Discussion on Smart Diversity Generation and Applicability for Different Protein Folds and Protein Properties
Exercises
FOCUSED MUTANT LIBRARY DESIGN
Classification of Methods for Single and Multi-Site Directed Mutagenesis
Performance and Comparison
Computational Tools for Focused Mutant Library Design
Discussion on Diversity Generation and Applicability for Specific Protein Properties
Exercises
SCREENING TECHNOLOGIES
General Consideration
Formats, Throughput and the Screening "Catastrophe"
Microtiter Plate Based Systems
Agar Plate Based Systems
Flow Cytometry Based Systems
Microfluidic Devices Based Systems
In Vitro Translation in Microcompartments/Emulsions
General Screening Platforms
General Lessons on Library Size and Obtained Property Improvements
Exercises
GENERAL LESSONS OF PROTEIN EVOLUTION
The Diversity and Screening "Catastrophes"
Thermal Stability
Activity Towards Non-Natural and Natural Substrates
Selectivity & One Catalyst/Many Substrate Concept
Substrate/Product Concentration
Solvent Resistance (Organic Solvents/Ionic Liquids)
pH-Resistance
Salt and Detergent Resistance
Alternative Cofactor Systems
Summary and Principles for Individual Property Improvements
Exercises
CHALLENGES, BARRIERS AND PROMISING STRATEGIES FOR DIRECTED EVOLUTION
Diversity Generation
Computational Simulation
Throughput Technologies and Selection Systems
Natural Evolution vs Directed Evolution
Exercises