Highlights•Optimization plateaus are common when engineering enzymes for higher
catalytic efficiency.•These plateaus relate to fundamental properties of evolutionary fitness …

Naturally occurring enzymes are remarkable biocatalysts with numerous potential
applications in industry and medicine. However, many of their catalyst properties often need …

Applied biocatalysis is driven by environmental and economic incentives for using enzymes
in the synthesis of various pharmaceutical and industrially important chemicals. Protein …

Directed evolution has now been used for over two decades as an alternative to rational
design for protein engineering. Protein function, however, is complex, and modifying …

The free energy landscape concept that describes enzymes as an ensemble of differently
populated conformational sub-states in dynamic equilibrium is key for evaluating enzyme …

Current challenges and promises of white biotechnology encourage protein engineers to
use a directed evolution approach to generate novel and useful biocatalysts for various sets …

Current computational tools to assist experimentalists for the design and engineering of
proteins with desired catalytic properties are reviewed. The applications of these tools for de …

The last decade has seen a dramatic increase in the utilization of enzymes as green and
sustainable (bio) catalysts in pharmaceutical and industrial applications. This trend has to a …

Many research groups successfully rely on whole-gene random mutagenesis and
recombination approaches for the directed evolution of enzymes. Recent advances in …

By constructing mutant libraries and utilizing high-throughput screening methods, directed
evolution has emerged as the most popular strategy for protein design nowadays. In the past …