title: [A novel bacterial cell-surface display system based on NCgl1221 from Corynebacterium glutamicum].
abstract: 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 beta-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 beta-amylase. Finally, we analyzed the activity of beta-amylase and starch hydrolization in order to determine whether the displayed beta-amylase has the activity or not.
  
  
  RESULTS
  The fusion protein was successfully expressed in E. coli, and the active beta-amylase was displayed on the cell surface by fusing it to the C terminus of the anchor. The recombinant strain displaying beta-amylase can utilize soluble starch in the medium.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Pharmacology, Nantong University Medical College, Nantong 226001, China. juanwenyy@yahoo.com.cn
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Yao
authors.forename: Wenjun
authors.initials: W
authors.lastname: Fan
authors.forename: Xiaole
authors.initials: X
authors.lastname: Xu
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Xiaozhao
authors.initials: X
authors.lastname: Deng
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D048230:Corynebacterium glutamicum / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D001614:beta-Amylase / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D001614:beta-Amylase