Improving enantioselectivity of lipase from Candida rugosa by carrier-bound and carrier-free immobilization

Publication date: Available online 12 May 2016 Source:Journal of Molecular Catalysis B: Enzymatic Author(s): Susana Velasco-Lozano, Fernando López-Gallego, Javier Rocha-Martin, José Manuel Guisán, Ernesto Favela-Torres The enantioselectivity of carrier-bound and carrier-free immobilized lipase from Candida rugosa (CRL) was studied. CRL was immobilized in six agarose-based carriers functionalized with different reactive groups and in two different CRL cross-linked aggregates. Both, activity and enantioselectivity of all the immobilized lipase preparations were evaluated with different racemic esters under different reaction conditions (temperature, pH and solvent polarity). A strong effect of reaction media and immobilization protocol on enzyme activity and selectivity was found. Enzyme immobilization and reaction engineering allowed us obtaining the best immobilization protocol and reaction conditions to achieve high activity and enantioselectivty of CRL as heterogeneous catalyst. CRL immobilized on agarose-based carrier activated with primary amino groups preferentially hydrolyzed (S)-phenylethyl acetate with E >200 under pH 7, 4°C and 30% of acetonitrile. On the other hand, CRL aggregated and cross-linked through their carboxylic groups preferentially hydrolyzed the (S)-isomer of ethyl 2-hydroxy-4-phenylbutyrate with an E =39 under pH 5, 4°C and 30% of acetonitrile. This work demonstrates the success of the combinatorial enzyme engineering for the ...

Source: Journal of Molecular Catalysis B: Enzymatic - May 12, 2016 Category: Biochemistry Source Type: research