From detergent additive to semisynthetic peroxidase-simplified and up-scaled synthesis of seleno-subtilisin

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