Engineering the Enantioselectivity ofB. subtilis

A novel genetic selection system for improved enantioselectivity of Bacillus subtilis lipase A.

In directed evolution experiments, success often depends on the efficacy of screening or selection methods. Genetic selections have proven to be extremely valuable for evolving enzymes with improved catalytic activity, improved stability, or with altered substrate specificity. In contrast, enantioselectivity is a difficult parameter to select for. In this study, we present a successful strategy...

Control of lipase enantioselectivity by engineering the substrate binding site and access channel.

Lipase from Burkholderia cepacia (BCL) has proven to be a very useful biocatalyst for the resolution of 2-substituted racemic acid derivatives, which are important chiral building blocks. Our previous work showed that enantioselectivity of the wild-type BCL could be improved by chemical engineering of the substrate's molecular structure. From this earlier study, three amino acids (L17, V266 and...

Understanding and Engineering the Enantioselectivity of Candida antarctica Lipase B towards sec-Alcohols

Improvements of enzyme activity and enantioselectivity via combined substrate engineering and covalent immobilization.

Esterases, lipases, and serine proteases have been applied as versatile biocatalysts for preparing a variety of chiral compounds in industry via the kinetic resolution of their racemates. In order to meet this requirement, three approaches of enzyme engineering, medium engineering, and substrate engineering are exploited to improve the enzyme activity and enantioselectivity. With the hydrolysis...

Enantioselectivity of haloalkane dehalogenases and its modulation by surface loop engineering.

Enzymes are widely used for the synthesis of pharmaceuticals, agrochemicals, and food additives because they can catalyze enantioselective transformations. Understanding the molecular basis of enzyme–substrate interactions that contribute to enantioselectivity is important for constructing selective enzymes by protein engineering. Up to now, emphasis has been on reactions such as lipaseor ester...