Tamaru Y, Araki T, Amagoi H, Mori H, Morishita T
Faculty of Bioresources, Mie University, Japan.
Appl Environ Microbiol. 1995 Dec;61(12):4454-8. doi: 10.1128/aem.61.12.4454-4458.1995.
A beta-mannanase (EC 3.2.1.78) from Vibrio sp. strain MA-138 was purified by ammonium sulfate precipitation and several chromatographic procedures including gel filtration, adsorption, and ion-exchange chromatographies. The final ion-exchange chromatography Mono Q yielded one major active fraction and three minor active fractions. The major active fraction was purified to homogeneity on the basis of native polyacrylamide gel electrophoresis (PAGE). This purified enzyme was identified as a glycoprotein by periodic acid-Schiff staining and a monomeric protein with a molecular mass of 49 kDa by sodium dodecyl sulfate-PAGE. The pI of the enzyme was 3.8. The purified enzyme exhibited maximal activity at pH 6.5 and 40 degrees C and hydrolyzed at random the internal beta-1,4-mannosidic linkages in beta-mannan to give various sizes of oligosaccharides. The first 20 N-terminal amino acid sequence of the purified enzyme showed high homology with the N-terminal region of beta-mannanase from Streptomyces lividans 66.
通过硫酸铵沉淀以及包括凝胶过滤、吸附和离子交换色谱在内的多种色谱方法，对来自弧菌属MA - 138菌株的一种β - 甘露聚糖酶（EC 3.2.1.78）进行了纯化。最后的离子交换色谱Mono Q产生了一个主要活性组分和三个次要活性组分。基于天然聚丙烯酰胺凝胶电泳（PAGE），主要活性组分被纯化至同质。通过高碘酸 - 席夫染色，该纯化酶被鉴定为糖蛋白，通过十二烷基硫酸钠 - PAGE被鉴定为分子量为49 kDa的单体蛋白。该酶的pI为3.8。纯化后的酶在pH 6.5和40℃时表现出最大活性，并随机水解β - 甘露聚糖中的内部β - 1,4 - 甘露糖苷键，产生各种大小的寡糖。纯化酶的前20个N端氨基酸序列与来自变铅青链霉菌66的β - 甘露聚糖酶的N端区域具有高度同源性。
