Whether the pursuit is commercially motivated or purely academic, engineering a novel biological catalyst is an enticing challenge. High-resolution protein structure analysis allows for rational alteration of enzyme function, yet many useful enzyme variants are the product of well-designed selection schemes or screening strategies. Enzyme Engineering: Methods and Protocols provides guidance to investigators wishing to create enzyme variants with desired properties. This detailed volume covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes. It also highlights the engineering of two difference types of rare-cutting endonucleases that show great potential in gene therapy applications: The newest development is the emergence of TAL effector nucleases or TALENs. Chapters describe newly developed technologies in sufficient detail so that each method can be practiced in a standard molecular biology laboratory. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible Enzyme Engineering: Methods and Protocols will be valuable for scientists with a budding interest in protein engineering as well as veterans looking for new approaches to apply in established discovery programs.

This book provides a comprehensive introduction to all aspects of enzyme engineering, from fundamental principles through to the state-of-the-art in research and industrial applications. It begins with a brief history, describing the milestones of advancement in enzyme science and technology, before going on to cover the fundamentals of enzyme chemistry, the biosynthesis of enzymes and their production. Enzyme stability and the reaction kinetics during enzymatic reactions are presented to show how enzymes function during catalysis and the factors that affect their activity. Methods to improve enzyme performance are also presented, such as cofactor regeneration and enzyme immobilization. The book emphasizes and elaborates on the performance and characteristics of enzymes at the molecular level. Finally, the book presents recent advances in enzyme engineering and some key industrial application of enzymes addressing the present needs of society. This book presents essential information not only for undergraduate and graduate students, but also for researchers in academia and industry, providing a valuable reference for the development of commercial applications of enzyme technology.