Biotechnology Letters

, Volume 30, Issue 6, pp 1025–1029

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

DOI: 10.1007/s10529-008-9656-y

Cite this article as:
Tanaka, Y., Doi, S., Kamiya, N. et al. Biotechnol Lett (2008) 30: 1025. doi:10.1007/s10529-008-9656-y

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 platePeptide tagProtein immobilizationSite-specific protein modificationTransglutaminase

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