Effect of the immobilization protocol on the properties of lipase B from Candida antarctica in organic media: Enantiospecifc production of atenolol acetate

In this work, we intend to check the effect of the immobilization protocol on the performance of lipase B from Candida antarctica (CalB) in organic medium. To this purpose, CalB has been immobilized on Eupergit C (EC) under different conditions and on EC partially modified with ethylenediamine (EDA), iminodiacetic acid (IDA) or metal chelate (IDA-Cu2+) and used for kinetic resolution of (R/S) 4-[2-hydroxy-3-[(1-methylethyl)amino]propoxy]benzeneacetamide (rac-atenolol). Enantiomeric resolution of atenolol was carried out by a transesterification reaction using vinyl acetate as acylant agent and an organic solvent as reaction medium. After a preliminary optimization of the reaction to obtain satisfactory yields, toluene was selected as the optimal solvent and the performances of the different CalB biocatalysts were compared. The R enantiomer was preferred in all cases but their performances were substantially different, with high differences in reaction rates, reaction yields in this kinetically controlled synthesis (EC-CalB gave a conversion of 76% while EC-IDA-Cu2+-CalB gave just a 27%), and enantiospecificities (EC-CalB gave an E value of 65 while EC-IDA-Cu2+-CalB gave a value of 13). Replacing toluene with hexane caused a decrease in enzyme activity, reaction yields and enantiospecificity of the reaction. It was remarkable that some preparations were much more sensitive to this solvent change than others. Considering that the activity decreased by less than 10% per reaction cycle, these differences are likely associated with the differences in the enzyme catalytic properties caused by the different immobilization protocols and not by inactivation of immobilized enzyme preparations during the reaction. These results confirmed that use of different immobilization protocols may be a powerful tool for altering enzyme properties when used in organic media. (C) 2011 Elsevier B.V. All rights reserved.