普通梨囊霉OTS1胞质几丁质酶的纯化及特性

Purification and characteristics of cytosolic chitinase from Piromyces communis OTS1.

作者信息

Sakurada M, Morgavi D P, Komatani K, Tomita Y, Onodera R

机构信息

Faculty of Agriculture, Miyazaki University, Japan.

出版信息

FEMS Microbiol Lett. 1996 Mar 15;137(1):75-8. doi: 10.1111/j.1574-6968.1996.tb08085.x.

DOI:10.1111/j.1574-6968.1996.tb08085.x

PMID:8935660

Abstract

A chitinase was purified from the cytosolic fraction of the anaerobic rumen fungus Piromyces communis OTS1 by affinity chromatography using regenerated chitin, gel filtration and chromatofocusing. The chitinase was most active at pH 6.2 and at 60 degrees C in a 20-min assay. The molecular mass of the purified protein was estimated by SDS-PAGE to be 42 kDa and its pI was 4.9. The enzyme activity, which was of the 'endo' type, was inhibited by Ag+, Hg2+ and allosamidin. N-Acetyl-beta-glucosaminidase and 'exo' type chitinase activity were absent from the purified preparation.

摘要

通过使用再生几丁质的亲和色谱法、凝胶过滤和色谱聚焦，从厌氧瘤胃真菌普通梨形霉OTS1的胞质部分中纯化出一种几丁质酶。在20分钟的测定中，该几丁质酶在pH 6.2和60℃时活性最高。通过SDS-PAGE估计纯化蛋白的分子量为42 kDa，其pI为4.9。该酶活性为“内切”型，受到Ag+、Hg2+和别洛沙米定的抑制。纯化制剂中不存在N-乙酰-β-葡萄糖苷酶和“外切”型几丁质酶活性。