枯草芽胞表面CotC和CotG共展示海藻糖合酶研究  

Surface Display Trehalose Synthase on Bacillus subtilis Spore by CotC and CotG

作　　者：刘洪玲 刘浩[1] 王腾飞[2] LIU Hongling;LIU Hao;WANG Tengfei(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China;School of Bioengineering,Qilu University of Technology(Shandong Academy of Sciences),Jinan 250353,China)

机构地区：[1]工业发酵微生物教育部重点实验室,天津科技大学生物工程学院,天津300457 [2]齐鲁工业大学(山东省科学院)生物工程学院,济南250353

出　　处：《天津科技大学学报》2020年第1期18-25,共8页Journal of Tianjin University of Science & Technology

基　　金：国家自然科学基金青年基金资助项目(31501413)

摘　　要：为了实现海藻糖合酶(TreS)在胞外的应用及避免胞内表达的不便利,构建了芽胞表面展示TreS的重组枯草芽胞杆菌.以枯草芽胞杆菌芽胞表面锚定蛋白CotC、CotG作为TreS融合展示蛋白,通过荧光共聚焦显微镜、Western blot、Dot blot及酶活性分析表明,TreS成功展示于枯草芽胞杆菌的芽胞表面,且由CotC和CotG共展示TreS芽胞表面酶活力大于CotC、CotG单独展示TreS酶活力之和.通过在TB培养基中培养,共展示重组芽胞表面酶活力(以芽胞干质量计)达到1511.6 U/g,表面展示分子数达到7.366×109.In order to realize the application of trehalose synthase(TreS)in the extracellular domain and avoid the inconvenient expression in cells,recombinant Bacillus subtilis WB800n was constructed to display TreS on spore surface.CotC and CotG were used as TreS display proteins.The results of fluorescence confocal microscopy,Western blot,Enzyme activity and Dot blot analysis showed that TreS was successfully displayed on the spore surface of Bacillus subtilis WB800n,and the enzyme activity of the co-displayed on the surface of spores by CotC and CotG exceeded the sum of the singly displayed on the spore surface.The surface enzyme activity of recombinant spore reached 1511.6 U/g(calculated by spore dry weight)and the number of displayed molecules reached 7.366×109 in the shaking flask culture using TB medium.

关 键 词：重组枯草芽胞杆菌 芽胞表面展示 基因敲除 海藻糖合酶 

分 类 号：Q814.2[生物学—生物工程]