| 引用本文： |
-
陈燕,裴建新,郭媛,林丽华,黄日波,庞浩.阴沟肠杆菌一个内切纤维素酶Cel8A的克隆和功能证实[J].广西科学,2014,21(1):17-21. [点击复制]
- CHEN Yan,PEI Jian-xin,GUO Yuan,LIN Li-hua,HUANG Ri-bo,PANG Hao.Cloning and Function Characteristics of an Endo-1,4-β-D-glucanase Cel8A Gene from Enterobacter cloacae[J].Guangxi Sciences,2014,21(1):17-21. [点击复制]
|
|
| |
|
|
| 本文已被：浏览 434次 下载 457次 |
码上扫一扫！ |
| 阴沟肠杆菌一个内切纤维素酶Cel8A的克隆和功能证实 |
|
陈燕1,2, 裴建新1, 郭媛1, 林丽华1,2, 黄日波1,2, 庞浩1,2
|
|
|
| (1.广西科学院, 非粮生物质酶解国家重点实验室, 国家非粮生物质能源工程技术研究中心, 广西生物质产业化工程院, 广西生物炼制重点实验室, 广西南宁 530007;2.广西大学生命科学与技术学院, 广西南宁 530004) |
|
| 摘要: |
| [目的]为了解决纤维素的酶水解问题,分析能水解纤维素的微生物的相关基因。[方法]从阴沟肠杆菌(Enterobacter cloacae)的基因组数据中挖掘纤维素酶相关基因,发现开放读码框Cel8A和纤维素酶基因序列具有高相似性。利用重组表达技术在大肠杆菌中对Cel8A基因进行表达,纯化重组蛋白质并对该蛋白质进行功能鉴定。[结果]Cel8A蛋白质能水解羧甲基纤维素钠,它具有β-1,4-内切葡聚糖酶的水解特性。Cel8A的最适反应pH值为7.0,最适温度为40℃。[结论]成功克隆表达阴沟肠杆菌的Cel8A基因,并证实该基因编码的蛋白质具有β-1,4-内切葡聚糖酶活性,为进一步研究阴沟肠杆菌的纤维素酶组成以及纤维素降解机理奠定了基础。 |
| 关键词: β-1,4-内切葡聚糖酶 酶学特性 阴沟肠杆菌 羧甲基纤维素钠 纤维素水解 |
| DOI： |
| 投稿时间：2013-07-29修订日期：2013-09-15 |
| 基金项目:广西科技合作与交流计划项目(桂科合1347004-1),广西科技攻关项目(桂科攻10123007-3),广西科学院基本科研业务费项目(12YJ25SW05,13YJ22SW),国家自然科学基金项目(21366007)资助。 |
|
| Cloning and Function Characteristics of an Endo-1,4-β-D-glucanase Cel8A Gene from Enterobacter cloacae |
|
CHEN Yan1,2, PEI Jian-xin1, GUO Yuan1, LIN Li-hua1,2, HUANG Ri-bo1,2, PANG Hao1,2
|
| (1.Guangxi Academy of Sciences, State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Industrialization Engineering Institute, Guangxi Key Laboratory of Biorefinery, Nanning, Guangxi, 530007, China;2.College of Life Science & Technology of Guangxi University, Nanning, Guangxi, 530004, China) |
| Abstract: |
| [Objective] The enzymatic hydrolysis of cellulose is a problem waiting for being solved.[Method] In this study, DNA sequence similar with endo-1, 4-β-D-glucanase was screened from the genome data of Enterobacter cloacae, where one DNA fragment Cel8A showed high similarity was found.This DNA was cloned, expressed, and the resulting protein was purified and its function characteristics were analyzed.[Result] This Cel8A could hydrolyze carboxymethyl cellulose sodium and showed the endo-1, 4-β-D-glucanase action characteristics.Its optimal pH was 7.0 and optimal temperature was 40℃.[Conclusion] In this work, an endo-1, 4-β-D-glucanase gene was successfully found and cloned from E.cloacae, which establishes the foundation for further study of the cellulase system and their working mechanism of E.cloacae. |
| Key words: endo-1,4-β-D-glucanase enzymatic characteristics Enterobacter cloacae carboxymethyl cellulose sodium cellulose hydrolysis |
|
|
|
|
|
