New strategies for enzyme stabilization involving molecular evolution and immobilization in mesoporous materials

• Takahashi Haruo

  Biotechnology Lab., Toyota Central R&D Labs. Inc., Japan

• Miyazaki-Imamura Chie

  Biotechnology Lab., Toyota Central R&D Labs. Inc., Japan

抄録

Heme containing peroxidase, for example manganese peroxidase (MnP), is easily inactivated by the hydrogen peroxide (H₂O₂) presented in the reaction. Here we extremely increased the H₂O₂ stability of MnP by molecular evolution and immobilization in mesoporous materials. A mutant MnP library containing three randomized amino acid residues located in the entry site of H₂O₂-binding pocket of MnP was evolved on a 384-well plate using SIMPLEX (single-molecule PCR-linked in vitro expression). The screening of more than 10⁴ samples independently expressed for improved H₂O₂ stability led to four positive mutants, the H₂O₂ stability of which was nine times higher than that of the wild-type. Immobilized MnP mutant in mesoporous material (FSM) showed the high H₂O₂ stability, more than 50 folds than wild type MnP. But the stability of immobilized wild type MnP was not improved so much as that for immobilized mutant MnP. [DOI: 10.1380/ejssnt.2005.207]

収録刊行物

• e-Journal of Surface Science and Nanotechnology

  e-Journal of Surface Science and Nanotechnology 3 207-212, 2005

  公益社団法人 日本表面真空学会

キーワード

• Biological aspect of nano-structures
• Catalysis
• Manganese

詳細情報

• CRID: 1390282680163571584
• NII論文ID: 130004933877
• DOI: 10.1380/ejssnt.2005.207
• ISSN: 13480391
• Web Site: http://www.jstage.jst.go.jp/article/ejssnt/3/0/3_0_207/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • Crossref
  • CiNii Articles
  • KAKEN
• 抄録ライセンスフラグ: 使用不可