文献类型：期刊文献

中文题名：枯草芽孢杆菌葡聚糖内切酶的克隆与表征

英文题名：Cloning and Characterization of an Endoglucanase from Bacillus sp.ECU0013

作者：邱立欢[1];李春秀[1];许建和[1]

机构：[1]华东理工大学生物反应器国家重点实验室,上海200237

年份：2011

卷号：37

期号：3

起止页码：265

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;Scopus;北大核心:【北大核心2008】；CSCD:【CSCD2011_2012】；

基金：国家自然科学基金(31071604)

语种：中文

中文关键词：枯草芽孢杆菌;葡聚糖内切酶;酶学性质;克隆;表征

外文关键词：Bacillus subtilis; endoglucanases; enzymatic properties; cloning; characterization

摘要：利用已公布的枯草芽孢杆菌基因组序列中假定的葡聚糖内切酶基因进行引物设计,从菌株Bacillussp.ECU0013中克隆表达得到重组葡聚糖内切酶BsEG。经镍柱纯化后进行酶学性质表征,其最适反应pH约为6.0;最适反应温度为50℃,具有较好的热稳定性,50℃时半衰期达101h。Km值为20.1 g/L,相应的Vmax值为0.075 g/(L·min)。并且初步对酶BsEG的底物特异性及对纤维素的吸附性进行了探究。
Based on a putative endoglucanase gene of Bacillus subtilis,we designed the primers and cloned a recombinant EG gene from Bacillus sp.ECU0013.Enzyme BsEG was successfully overexpressed in Escherichia coli and purified to homogeneity by Ni-NTA column.The maximal activity could be reached when pH was about 6.0 and reaction temperature was 50 ℃.The half-life time of EGbs at 50 ℃ was up to 101 h.Km and Vmax were 20.1 g/L and 0.075 g/（L·min）,respectively.Futhermore,the substrate specificity and cellulose absorption behavior of BsEG were studied.