来自特异青霉IFO 7237的细胞内α-D-木糖苷酶的纯化与特性分析

Purification and characterization of an intracellular alpha-D-xylosidase from Penicillium wortmannii IFO 7237.

作者信息

Matsuo M, Seki T, Mitsuishi Y, Shoun H, Nakahara T

机构信息

Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.

出版信息

Biosci Biotechnol Biochem. 1996 Feb;60(2):341-3. doi: 10.1271/bbb.60.341.

DOI:10.1271/bbb.60.341

PMID:9063986

Abstract

Intracellular alpha-D-xylosidase from Penicillium wortmannii IFO 7237 was obtained by grinding the mold with almina in phosphate buffer, and the cell-free extract was purified to a homogeneous state on SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was estimated to be 290,000 by gel filtration chromatography (Superdex 200) and 73,000 was obtained by SDS-PAGE. The purified alpha-D-xylosidase had an isoelectric point at pH 5.0. The optimum activity for the enzyme was found to be at pH 6.5 and 45 degrees C. The enzyme showed a hydrolytic activity on p-NO2-phenyl-alpha-D-xylopyranoside (alpha-p-NPX) while methyl-alpha-D-xylopyranoside (alpha-MX) was not hydrolyzed at all. It also showed lower activity for xyloglucan oligosaccharides. The apparent Km values of the enzyme for alpha-p-NPX and isoprimeverose were 1.9 mM and 50 mM, respectively.

摘要

来自特异青霉IFO 7237的细胞内α-D-木糖苷酶是通过在磷酸盐缓冲液中用氧化铝研磨该霉菌而获得的，无细胞提取物在SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）上被纯化至均一状态。通过凝胶过滤色谱法（Superdex 200）估计分子量为290,000，通过SDS-PAGE得到的分子量为73,000。纯化的α-D-木糖苷酶的等电点为pH 5.0。发现该酶的最佳活性在pH 6.5和45℃。该酶对对硝基苯基-α-D-吡喃木糖苷（α-p-NPX）表现出水解活性，而甲基-α-D-吡喃木糖苷（α-MX）完全不被水解。它对木葡聚糖寡糖也表现出较低的活性。该酶对α-p-NPX和异麦芽糖苷的表观Km值分别为1.9 mM和50 mM。