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    邱立欢, 李春秀, 许建和. 枯草芽孢杆菌葡聚糖内切酶的克隆与表征[J]. 华东理工大学学报(自然科学版), 2011, (3): 265-267.
    引用本文: 邱立欢, 李春秀, 许建和. 枯草芽孢杆菌葡聚糖内切酶的克隆与表征[J]. 华东理工大学学报(自然科学版), 2011, (3): 265-267.
    QIU Li-huan, LI Chun-xiu, XU Jian-he. Cloning and Characterization of an Endoglucanase from Bacillus sp. ECU0013[J]. Journal of East China University of Science and Technology, 2011, (3): 265-267.
    Citation: QIU Li-huan, LI Chun-xiu, XU Jian-he. Cloning and Characterization of an Endoglucanase from Bacillus sp. ECU0013[J]. Journal of East China University of Science and Technology, 2011, (3): 265-267.

    枯草芽孢杆菌葡聚糖内切酶的克隆与表征

    Cloning and Characterization of an Endoglucanase from Bacillus sp. ECU0013

    • 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Bacillus sp. 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101 h。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。

       

      Abstract: 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.

       

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