Selectivity paradigm in lipase reactions: correlating actual and observed regioselectivity in hydrolysis of unsymmetrical diacetates

Abstract

The first simple method using labeling technique has been developed to confirm the actual and observed selectivities in the enzymatic hydrolysis of unsymmetrical diacetates and to measure the enzyme selectivity efficiency. The simple method consists of enzymatic hydrolysis of unsymmetrical diacetate followed by labeling of the hydroxyacetate formed with CD₃COOD/DCC and enzymatic rehydrolysis of the labeled compound under the identical set of reaction conditions to estimate the amount of label retained by ¹H NMR spectroscopy. The amount of label lost directly indicates the extent of regioselective action of the enzyme.