Enzymes have been used as catalysts in organic chemistry and biotechnology for more than a century. But enzyme catalyzed reactions often encounter limitations, which can be overcome by engineering the enzyme and its catalytic profile. This can be achieved by directed evolution methods, which mimic evolutionary pressure and enable the identification of improved enzyme variants in several cycles of mutagenesis, expression and screening. In this book, Manfred Reetz gives an overview about basic strategies in directed evolution and introduces common gene mutagenesis, screening and selection methods. Emphasis is placed on methodology development to maximize efficiency, reliability and speed of such experiments and to provide guidelines for efficient protein engineering. Reetz highlights the application of directed evolution experiments to address limitations in the field of enzyme selectivity, substrate scope, activity and robustness. Recent developments are critically reviewed and an outlook is given to applications in the field of organic synthesis, e.g. to produce chiral pharmaceuticals, fragrances and plant protecting agents. As part of the critical review, Reetz discusses case studies and concludes with lessons learned from directed evolution experiments.

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