From detergent additive to semisynthetic peroxidase—simplified and up-scaled synthesis of seleno-subtilisin
✍ Scribed by Dietmar Häring; Peter Schreier
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 79 KB
- Volume
- 59
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
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. Activa- tion 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.