Solubilization, purification and characterization of D-alanine dehydrogenase from Pseudomonas aeruginosa and effects of solubilization on its properties |
| |
Authors: | A M Magor W A Venables |
| |
Affiliation: | 1. TEDA School of Biological Sciences and Biotechnology, Nankai University, Tianjin, China;2. The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Tianjin, China;3. N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia |
| |
Abstract: | D-alanine dehydrogenase, an inducible, membrane associated enzyme of Pseudomonas aeruginosa was solubilized from envelope preparations by treatment with Triton X-100 and purified 31-fold in the presence of 0.05% Triton X-100 to 60% homogeneity. Gel electrophoresis indicated the presence of a single subunit of approximately 49,000 molecular weight. The enzyme contained FAD, and absorption spectra were typical of an iron-sulfur flavoprotein. Solubilization produced significant changes in some properties of the enzyme: solubilized enzyme showed increased affinity for D-alanine; a broader substrate specificity; and increased temperature sensitivity, compared with the membrane associated form. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|