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

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

研究成果: Contribution to journalArticle査読

22 被引用数 (Scopus)

抄録

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.

本文言語英語
ページ(範囲)613-618
ページ数6
ジャーナルEnzyme and Microbial Technology
35
6-7
DOI
出版ステータス出版済み - 12 1 2004

All Science Journal Classification (ASJC) codes

  • バイオテクノロジー
  • バイオエンジニアリング
  • 生化学
  • 応用微生物学とバイオテクノロジー

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