玉米细菌性枯萎病菌EGase内切葡聚糖酶蛋白纯化研究初探

Initial studies of Endoglucanase purification from Pantoea stewartii subsp. stewartii

中文关键词:  玉米细菌性枯萎病菌  内切葡聚糖酶  蛋白纯化

英文关键词:Pantoea stewartii subsp. stewartii  endoglucanase  protein purification

中文摘要:

      目的 研究玉米细菌性枯萎病菌EGase内切葡聚糖酶蛋白纯化的方法。方法 以玉米细菌性枯萎病菌(Pantoea stewartii subsp. stewartii)菌株为材料, 分离纯化其内切葡聚糖酶Egase。制备EGase浓缩液, 浓缩液经SephradexTM G-75凝胶过滤层析和DEAE-Sephorose Fast Flow阴离子交换柱层析等提纯步骤, 获得了凝胶电泳均一的内切葡聚糖酶。结果 经变性聚丙烯酰胺凝胶电泳检测为一条电泳带, 纯化后的EGase是单体蛋白, 分子量约为72.3 kDa。Egase酶反应的最适温度是60 ℃, 最适pH为5.0。结论 本研究从玉米细菌性枯萎病菌中分离得到了一种新的内切葡聚合糖酶, 对其部分性质进行了表述, 为后续EGase基因的克隆及表达研究提供了研究基础。

英文摘要:

      Objective To investigate the purification method of endoglucanase (EGase) from Pantoea stewartii subsp. stewartii. Methods With the material of Pantoea stewartii subsp. stewartii. strain, EGase fermented liquid was prepared, and then eluted using SephadexTM G-75 and on DEAE-Sepharose Fast Flow anion exchange. SDS-PAGE was used to determine the EGase activity after purification. Results Endoglucanase was purified from Pantoea stewartii subsp. stewartii. It was monomer protein and its molecular size was 72.3 kDa. The optimum reaction temperature of endoglucanase was 60 ℃ and the optimum pH was 5.0. Conclusion Endoglucanase was purified firstly from Pantoea stewartii subsp. stewartii and its properties were described. This study will provide an important base for cloning and expression of endoglucanase gene research.