%0 Journal Article
%T A novel bacterial cell-surface display system based on NCgl1221 from Corynebacterium glutamicum<br>基于谷氨酸棒杆菌NCgl1221蛋白的新型细菌表面展示系统
%A Yao Wenjuan
%A Fan Wenjun
%A Xu Xiaole
%A Zhang Wei
%A Deng Xiaozhao
%A <br>Yao W
%A Fan W
%A Xu X
%A Zhang W
%A Deng X
%J 微生物学报
%D 2012
%I 
%X Objective]To develop a novel Escherichia coli cell surface display system by using C-terminally truncated NCgl1221 as the anchoring protein,which greatly enriched or optimized the bacterial displayed systems.Methods]We amplified the sequence of C-terminally truncated NCgl1221 and β-amylase,and constructed the fusion expression vector.Then we transformed the recombinant plasmids PET-NA and PET-28a into Rosetta(DE3)pLysS.The fusion protein expression was induced by IPTG and identified by SDS-PAGE and Western blot analysis.The IPTG induced strains were immunostained and investigated by fluorescence microscope and flow cytometry to detect the displayed β-amylase.Finally,we analyzed the activity of β-amylase and starch hydrolization in order to determine whether the displayed β-amylase has the activity or not.Results]The fusion protein was successfully expressed in E.coli,and the active β-amylase was displayed on the cell surface by fusing it to the C terminus of the anchor.The recombinant strain displaying β-amylase can utilize soluble starch in the medium.Conclusion]A novel E.coli surface display system by using C-terminally truncated NCgl1221 as the anchor motif was successfully developed.The active enzyme with a molecular size of 56 kDa was displayed on E.coli by this system,which provided the basis for the application of the system in whole-cell biocatalyst or biosorbent.
%K Corynebacterium glutamicum
%K β-amylase
%K cell-surface display
%K NCgl1221<br>谷氨酸棒杆菌(Corynebacterium
%K glutamicum)
%K β-淀粉酶
%K 细菌表面展示
%K NCgl1221
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=6D2AF9ACA55ECD33C39CA02CFD0FC668&yid=99E9153A83D4CB11&vid=286FB2D22CF8D013&iid=0B39A22176CE99FB&sid=6425DAE0271BB751&eid=DD74772618543076&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=0