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Development of novel cell surface display in Corynebacterium glutamicum using porin

  • Applied Genetics and Molecular Biotechnology
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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.

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Acknowledgements

This work was supported by a grant-in-aid for JSPS Fellows (20859 to T. Tateno) and by Special Coordination Funds for Promoting Science and Technology, Creation of Innovation Centers for Advanced Interdisciplinary Research Areas (Innovative Bioproduction Kobe), MEXT, Japan.

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Authors and Affiliations

  1. Department of Molecular Science and Material Engineering, Graduate School of Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

    Toshihiro Tateno

  2. Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

    Kazuki Hatada

  3. Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

    Tsutomu Tanaka & Hideki Fukuda

  4. Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, 657-8501, Japan

    Akihiko Kondo

Authors
  1. Toshihiro Tateno
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  2. Kazuki Hatada
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  3. Tsutomu Tanaka
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  4. Hideki Fukuda
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  5. Akihiko Kondo
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Correspondence to Akihiko Kondo.

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Tateno, T., Hatada, K., Tanaka, T. et al. Development of novel cell surface display in Corynebacterium glutamicum using porin. Appl Microbiol Biotechnol 84, 733–739 (2009). https://doi.org/10.1007/s00253-009-2021-9

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  • DOI: https://doi.org/10.1007/s00253-009-2021-9

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