Enzymatic site-selectivity enabled by structure-guided directed evolution

Structure-guided directed evolution of alkenyl and arylmalonate decarboxylases.

The asymmetric decarboxylation of prochiral a-disubstituted malonic acids is an attractive route to produce homochiral asubstituted carboxylic acids. Various metal-catalysed and organocatalytic methods have been explored for related enantioselective decarboxylative protonation (EDP) reactions. However, the synthetic EDPmethods can only deliver chiral carboxylic acids or esters with moderate ena...

Directed evolution strategies for improved enzymatic performance

The engineering of enzymes with altered activity, specificity and stability, using directed evolution techniques that mimic evolution on a laboratory timescale, is now well established. However, the general acceptance of these methods as a route to new biocatalysts for organic synthesis requires further improvement of the methods for both ease-of-use and also for obtaining more significant chan...

Directed Evolution of Glutathione Transferases Guided by Multivariate Data Analysis

Engineering of a target site-specific recombinase by a combined evolution- and structure-guided approach

Site-specific recombinases (SSRs) can perform DNA rearrangements, including deletions, inversions and translocations when their naive target sequences are placed strategically into the genome of an organism. Hence, in order to employ SSRs in heterologous hosts, their target sites have to be introduced into the genome of an organism before the enzyme can be practically employed. Engineered SSRs ...

Design by Directed Evolution

The stunning array of features and functions exhibited by proteins in nature should convince most scientists of the power of evolutionary design processes. Natural selection acting on populations over long periods of time has generated a vast number of proteins ideally suited to their biological functions. When we try to recruit these remarkable molecular machines for new taskssfrom serving as ...