Molecular Engineering of Enzymes

This chapter provides an overview of the different approaches for the design, redesign, and identification of enzymes with improved or new functionalities. It explores industrially relevant biocatalysts such as lipases, cellulases, and the cytochrome P450 to demonstrate the applicability of different protein engineering approaches. Protein engineering has proven successful in altering the properties of enzymes either through rational design or directed evolution. Rational design relies on structure- and usually mechanistic-guided predictions about the mutations that may be needed to modify a given enzyme property. The chapter describes computational toolboxes and molecular methods developed to create focused libraries. Molecular biology has been successfully applied in the case of a number of enzymes aiming for activity, specificity, and stability improvements with remarkable results in some cases. Efficient design of tailored activities remains a challenging test for the current understanding of how proteins fold and function and defies current knowledge of enzyme catalysis.