Biotechnology Letters

, Volume 30, Issue 6, pp 1025–1029 | Cite as

A chemically modified glass surface that facilitates transglutaminase-mediated protein immobilization

  • Yusuke Tanaka
  • Satoshi Doi
  • Noriho Kamiya
  • Noriyuki Kawata
  • Shinji Kamiya
  • Kenichi Nakama
  • Masahiro Goto
Original Research Paper

Abstract

An amino-modified glass surface for enzymatic protein immobilization by microbial transglutaminase (MTG) was developed. Diamine substrates with secondary amino groups in the linker moiety, like triethylenetetramine (TETA), exhibited at most a 2-fold higher reactivity in the MTG-catalyzed reaction compared to those with the alkyl linker. A 96-well glass plate was subsequently modified with selected diamine substrates. Validation of the modified surface by enzymatic immobilization of enhanced green fluorescent protein tagged with a glutamine donor-substrate peptide (LLQG) of MTG revealed that the protein loading onto the TETA-modified glass surface was approximately 15-fold higher than that on the unmodified one.

Keywords

96-Well glass plate Peptide tag Protein immobilization Site-specific protein modification Transglutaminase 

Notes

Acknowledgments

We are grateful to Ajinomoto Co., Inc. for providing the sample of MTG. The present work was supported by the Industrial Technology Research Grant Program in 2006 from the New Energy and Industrial Technology Development Organization (NEDO) of Japan (to N.K.).

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Copyright information

© Springer Science+Business Media B.V. 2008

Authors and Affiliations

  • Yusuke Tanaka
    • 1
  • Satoshi Doi
    • 1
  • Noriho Kamiya
    • 1
    • 2
  • Noriyuki Kawata
    • 3
  • Shinji Kamiya
    • 3
  • Kenichi Nakama
    • 3
  • Masahiro Goto
    • 1
    • 2
  1. 1.Department of Applied Chemistry, Graduate School of EngineeringKyushu UniversityFukuokaJapan
  2. 2.Center for Future ChemistryKyushu UniversityFukuokaJapan
  3. 3.New Products & Buisiness Development DepartmentNippon Sheet Glass Co., Ltd.TsukubaJapan

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