铜绿假单胞菌D-丙氨酸脱氢酶的增溶、纯化及特性鉴定以及增溶对其性质的影响。

Solubilization, purification and characterization of D-alanine dehydrogenase from Pseudomonas aeruginosa and effects of solubilization on its properties.

作者信息

Magor A M, Venables W A

出版信息

Biochimie. 1987 Jan;69(1):63-9. doi: 10.1016/0300-9084(87)90272-0.

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.

摘要

D-丙氨酸脱氢酶是铜绿假单胞菌的一种可诱导的膜相关酶，通过用 Triton X-100 处理从包膜制剂中溶解出来，并在 0.05% Triton X-100 存在下纯化了 31 倍，达到 60% 的纯度。凝胶电泳表明存在一个分子量约为 49,000 的单一亚基。该酶含有黄素腺嘌呤二核苷酸（FAD），吸收光谱是铁硫黄素蛋白的典型特征。溶解使该酶的一些性质发生了显著变化：与膜相关形式相比，溶解的酶对 D-丙氨酸的亲和力增加；底物特异性更广泛；温度敏感性增加。