Semi-rational approaches to engineering enzyme activity: combining the benefits of directed evolution and rational design.
Many research groups successfully rely on whole-gene random mutagenesis and recombination approaches for the directed evolution of enzymes. Recent advances in enzyme engineering have used a combination of these random methods of directed evolution with elements of rational enzyme modification to successfully by-pass certain limitations of both directed evolution and rational design. Semi-rational approaches that target multiple, specific residues to mutate on the basis of prior structural or functional knowledge create 'smart' libraries that are more likely to yield positive results. Efficient sampling of mutations likely to affect enzyme function has been conducted both experimentally and, on a much greater scale, computationally, with remarkable improvements in substrate selectivity and specificity and in the de novo design of enzyme activities within scaffolds of known structure.
Chica
Roberto A
RA
Département de chimie, Université de Montréal, CP 6128, Succursale Centre-Ville, Montréal, Québec, H3C 3J7, Canada.
Doucet
Nicolas
N
Pelletier
Joelle N
JN
D020650:Combinatorial Chemistry Techniques; D019295:Computational Biology; D019020:Directed Molecular Evolution; D004798:Enzymes / Q000235:genetics / Q000502:physiology*; D015202:Protein Engineering
D004798:Enzymes