An enzymatic strategy for site-specific immobilization of functional proteins using microbial transglutaminase

Jo Tominaga, Noriho Kamiya, Satoshi Doi, Hirofumi Ichinose, Masahiro Goto

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

A novel strategy for site-specific immobilization of recombinant proteins was investigated using microbial transglutaminase (MTG). Alkaline phosphatase (AP) was selected as a model protein and tagged with a short peptide (MKHKGS) at the N-terminus to provide a reactive Lys residue for MTG. On the other hand, casein, a well-known substrate for MTG, was chemically attached onto a polyacrylic resin to provide reactive Gln residues for the enzymatic immobilization of the recombinant AP. As a result, we succeeded in MTG-mediated functional immobilization of the recombinant AP onto casein-coated polyacrylic resin. It was found that the immobilized AP prepared using MTG exhibited much higher specific activity than that prepared by chemical modification. Moreover, enzymatic immobilization gave an immobilized formulation with higher stability upon repeated use than that obtained by physical adsorption. Use of this ability of MTG in posttranslational protein modification will provide us with a benign, site-specific immobilization method for functional proteins.

Original languageEnglish
Pages (from-to)613-618
Number of pages6
JournalEnzyme and Microbial Technology
Volume35
Issue number6-7
DOIs
Publication statusPublished - Dec 1 2004

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology

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