Applied Microbiology and Biotechnology

, Volume 84, Issue 4, pp 733–739 | Cite as

Development of novel cell surface display in Corynebacterium glutamicum using porin

  • Toshihiro Tateno
  • Kazuki Hatada
  • Tsutomu Tanaka
  • Hideki Fukuda
  • Akihiko Kondo
Applied Genetics and Molecular Biotechnology

Abstract

We have developed a novel cell surface display in Corynebacterium glutamicum using porin proteins as anchor proteins. Porins are localized at C. glutamicum mycolic acid layer and exist as a hexamer. We used α-amylase from Streptococcus bovis 148 (AmyA) as a model protein to be displayed on the C. glutamicum cell surface. AmyA was fused to the C terminus of the porins PorB, PorC, or PorH. Expression vectors using fused proteins under the control of the cspB promoter were constructed and introduced into the C. glutamicum Cm strain. Immunostaining microscopy and flow cytometric analysis revealed that PorB-AmyA, PorC-AmyA, and PorH-AmyA were displayed on the C. glutamicum cell surface. AmyA activity was only detected in the cell fraction of C. glutamicum cells that displayed AmyA fused to PorB, PorC or PorH and AmyA activity was not detected in the supernatants of C. glutamicum culture broths after 72 h cultivation. Thus, we have demonstrated that C. glutamicum porins are very efficient anchor proteins for protein display in C. glutamicum.

Keywords

Corynebacterium glutamicum Cell surface display Porin 

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Toshihiro Tateno
    • 1
  • Kazuki Hatada
    • 2
  • Tsutomu Tanaka
    • 3
  • Hideki Fukuda
    • 3
  • Akihiko Kondo
    • 4
  1. 1.Department of Molecular Science and Material Engineering, Graduate School of Science and TechnologyKobe UniversityKobeJapan
  2. 2.Department of Chemical Science and Engineering, Faculty of EngineeringKobe UniversityKobeJapan
  3. 3.Organization of Advanced Science and TechnologyKobe UniversityKobeJapan
  4. 4.Department of Chemical Science and Engineering, Graduate School of EngineeringKobe UniversityKobeJapan

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