FROM DETERGENT ADDITIVE TO SEMISYNTHETIC PEROXIDASE - SIMPLIFIED AND UP-SCALED SYNTHESIS OF SELENO-SUBTILISIN

A simplified and up-scaled synthesis of the semisynthetic peroxidase s eleno-subtilisin was developed. Highly purified to technical grade sub tilisin preparations from Bacillus licheniformis and Bacillus amyloliq uefaciens were applied as starting materials. Activation of Ser 221 wi th phenylmethanesulfonyl fluoride, nucleophilic substitution by sodium hydrogen selenide, and oxidation to the seleninic acid with hydrogen peroxide completed the chemical active-site modification. The reaction s were accomplished with a minimum of simple work-up procedures in 10 g scale. Kinetics and enantioselectivity of the preparations were test ed using 1-phenylethyl hydroperoxide. For the first time, an up-scaled synthesis of a semisynthetic enzyme is available. (C) 1998 John Wiley & Sons, Inc.