|本期目录/Table of Contents|

[1]杨立红,程仕伟,冯志彬,等.链霉菌壳聚糖酶的纯化及其酶学性质[J].生物加工过程,2013,11(03):52-58.[doi:10.3969/j.issn.1672-3678.2013.03.010]
 YANG Lihong,CHENG Shiwei,FENG Zhibin,et al.Purification chitosanase from Streptomyces hygroscopious and its catalytic properties[J].Chinese Journal of Bioprocess Engineering,2013,11(03):52-58.[doi:10.3969/j.issn.1672-3678.2013.03.010]
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链霉菌壳聚糖酶的纯化及其酶学性质()
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《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
11
期数:
2013年03期
页码:
52-58
栏目:
出版日期:
2013-05-30

文章信息/Info

Title:
Purification chitosanase from Streptomyces hygroscopious and its catalytic properties
文章编号:
1672-3678(2013)03-0052-07
作者:
杨立红程仕伟冯志彬周楠楠孔云红明永飞
鲁东大学 生命科学学院 微生物工程系,烟台 264025
Author(s):
YANG LihongCHENG ShiweiFENG ZhibinZHOU NannanKONG YunhongMing Yongfei
Institute of Microbial Engineering,College of Life Sciences,Ludong University,Yantai 264025,China
关键词:
酶学性质 壳聚糖酶 分离纯化 链霉菌
分类号:
Q814
DOI:
10.3969/j.issn.1672-3678.2013.03.010
文献标志码:
A
摘要:
分离纯化从烟台近海土壤筛选的链霉菌来源壳聚糖酶,并对其酶学性质进行研究。通过(NH4)2SO4分级沉淀分离得粗酶,透析后经Sephadex G-100柱纯化,得到2种壳聚糖酶(ChA和ChB)。SDS-聚丙烯酰胺凝胶电泳及Sephadex G-75凝胶过滤确定ChA的相对分子质量,研究ChA的最适底物水解条件、热稳定性、水解动力学及金属离子对酶活性影响。结果表明:ChA为单亚基蛋白,相对分子质量为4.16×104,在220和280 nm处呈现两个紫外吸收峰,催化水解壳聚糖的最适pH为5.0~5.5,最适温度为55 ℃。热稳定性实验表明:30 ℃温育1 h后酶活为初始酶活的33.3%,40 ℃温育1 h后酶活为初始酶活的22.2%。ChA的酶促反应初速率为6.2×10-3 μmol/(mL·min), Vmax为0.318 μmol/(mL·min),Km为1×10-2 mg/mL,且对底物表现相对专一性。K+、Na+、Li+、 Mg2+、Ca2+、Ba2+ Zn2+、Cu2+和Co2+对ChA活力均表现为抑制作用,过渡金属离子Mn2+对酶有激活作用,重金属离子Hg2+、Ag+、Cd2+ 和Pb2+对酶均有较强的抑制作用。Mn2+和Zn2+的动力学研究表明,Mn2+对酶为混合型激活作用,Zn2+对酶为竞争性抑制作用。

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备注/Memo

备注/Memo:
收稿日期:2011-12-02
基金项目:山东省高等学校科技计划(J12LD02); 鲁东大学科研基金(LY20083305)
作者简介:杨立红(1959—),女,辽宁沈阳人,教授,研究方向:生物分离工程,E-mail:swbiotech@yeah.net.
更新日期/Last Update: 2013-03-30