Effect of the immobilization protocol in the activity, stability, and enantioslectivity of Lecitase® Ultra

Lecitase® Ultra is a commercial enzyme preparation designed for degumming of oils. In this paper, we show that this enzyme increased its hydrolytic activity in the presence of low concentrations of detergents in hydrolytic reactions (e.g., 150-fold using 0.01% hexadecyltrimethylammonium bromide). Moreover, the enzyme becomes adsorbed on octyl agarose at low ionic strength, increasing the activity by a 13-fold factor. The enzyme adsorbed on octyl-agarose was desorbed by addition of detergents and immobilized on supports coated with polyethyleneimine (PEI) and in cyanogen bromide agarose. This immobilized preparations exhibited very different activity and enantioselectivity in the hydrolysis of (±)-methyl mandelate and (±)-2-O-butanoyl-2-phenylacetic acid. For example, the enzyme immobilized on octyl agarose yielded R-mandelic acid with an E value higher than 100, while the CNBr preparation gave an E value of 26, favoring the S isomer. The immobilized enzyme was quite stable at pH 5 and 7, and in the presence of organic solvents. Lecitase® Ultra seems to have very good prospects to be used as enantioselective biocatalyst in fine chemistry.