Site-directed mutagenesis of cysteine to threonine in Proteus vulgaris urease active site increases enzyme activity and stability |
| |
| Authors: | Joachim Jose Sven Lauter Martin A Stein |
| |
| Institution: | (1) Pharmaceutical and Medicinal Chemistry, University of the Saarland, P.O. Box, 151150, 66041 Saarbrücken, Germany;(2) Lion Bioscience AG, Waldhofer Str. 98, 69123 Heidelberg, Germany |
| |
| Abstract: | Cysteine-319 belongs to the flexible flap at the active site of Proteus vulgaris urease. Replacing this cysteine by threonine resulted in a 20-fold increase of specific activity. Temperature stability increased, susceptibility to inhibition by dipyridyl disulfide decreased, and pH optimum shifted from 8 to 6.9. K
m (35 to 12 mM) and Vmax (47.4 to 1.8  mol min–1) were substancially altered. Both variants of the enzyme were irreversibly inhibited by phenylmethanesulfonyl fluoride. |
| |
| Keywords: | active site Proteus vulgaris site-directed mutagenesis Staphylococcus xylosus urease |
| 本文献已被 SpringerLink 等数据库收录！ |
|
