Principles of Protein Engineering Fundamentals, Tools and Strategies

Comprehensive Reference Work Detailing Basic and Advanced Information Surrounding Protein Engineering

Protein engineering is a method of changing a protein sequence to achieve a desired result, such as a change in the substrate specificity or increased stability to the temperature, organic solvents, and/or extremes of pH or selectivity in catalytic reactions. Principles of Protein Engineering provides researchers the necessary fundamental knowledge about the protein engineering field and the rationale behind the techniques.

Written by two experts with significant first-hand experience in the field, the work provides insight on protein synthesis, expression, structure elucidation, bioinformatics, directed evolution, stability, screening, and rational design. To aid in reader comprehension, the text is enhanced by short comments, feedback and personal anecdotes from pioneers and leaders in the field. Sample ideas covered in the work include:

• The fundamental knowledge of protein synthesis and expression
• Basic knowledge about alignment, homology modelling, dynamics of structure, and modeling enzyme kinetics
• The main protein engineering strategies, with a focus on directed evolution and rational design
• Differences between screening and selection, plus the main techniques for high-throughput screening

Biochemists, biotechnologists, molecular biologists, and students in related programs of study can use Principles of Protein Engineering to attain both holistic and specific insight into the field of protein engineering and its numerous applications.

Ioannis Pavlidis is head of the Enzyme Technology Laboratory and Assistant Professor at the Department of Chemistry, University of Crete (GR). Having obtained his academic degrees from University of Ioannina (GR), he spent 4 years at the Department of Biotechnology and Enzyme Catalysis, University of Greifswald (DE) and later he established his own group at the University of Kassel (Germany).

Christin Peters leads the Biosystems Technology group, which concentrates on enzyme cascade reactions and process optimization of large-scale biotransformations. She studied biochemistry at the University of Greifswald.

Introduction and Retrospect to Protein Engineering

PART I. Fundamentals of Proteins and Protein Expression
Introduction to Proteins and Protein Conformation
Protein Expression and the Selection of the Optimal Host
Protein Structure Determination

PART II. Bioinformatic Tools for Guiding Protein Engineering
Sequence Databases
Protein Modelling
Enzyme Kinetics

PART III. Protein Engineering Approaches/Strategies
Introduction to Protein Engineering
Selecting Strategy According to the Goal of the Engineering Project
Directed Evolution/Random Mutagenesis
Rational and Computational Design
Engineering Protein Stability
De Novo Design of Novel Enzymes

PART IV. Screening Mutant Libraries with High-Throughput Methods
Screening versus Selection
Auxotrophy
Development of Plate Assays for Colony Screening
Photometric /
Fluorometric Assays
Different Compartmental Approaches