Hybridization of Modified-Heme Reconstitution and Distal Histidine Mutation to Functionalize Sperm Whale Myoglobin

Hideaki Sato, Takashi Hayashi, Tsutomu Ando, Yoshio Hisaeda, Takafumi Ueno, Yoshihito Watanabe

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

To modulate the physiological function of a hemoprotein, most approaches have been demonstrated by site-directed mutagenesis. Replacement of the native heme with an artificial prosthetic group is another way to modify a hemoprotein. However, an alternate method, mutation or heme reconstitution, does not always demonstrate sufficient improvement compared with the native heme enzyme. In the present study, to convert a simple oxygen storage hemoprotein, myoglobin, into an active peroxidase, we applied both methods at the same time. The native heme of myoglobin was replaced with a chemically modified heme 2 having two aromatic rings at the heme-propionate termini. The constructed myoglobins were examined for 2-methoxyphenol (guaiacol) oxidation in the presence of H2O2. Compared with native myoglobin, rMb(H64D·2) showed a 430-fold higher kcat/Km value, which is significantly higher than that of cytochrome c peroxidase and only 3-fold less than that of horseradish peroxidase. In addition, myoglobin-catalyzed degradation of bisphenol A was examined by HPLC analysis. The rMb(H64D·2) showed drastic acceleration (>35-fold) of bisphenol A degradation compared with the native myoglobin. In this system, a highly oxidized heme reactive species is smoothly generated and a substrate is effectively bound in the heme pocket, while native myoglobin only reversibly binds dioxygen. The present results indicate that the combination of a modified-heme reconstitution and an amino acid mutation should offer interesting perspectives toward developing a useful biomolecule catalyst from a hemoprotein.

Original languageEnglish
Pages (from-to)436-437
Number of pages2
JournalJournal of the American Chemical Society
Volume126
Issue number2
DOIs
Publication statusPublished - Jan 21 2004

Fingerprint

Sperm Whale
Myoglobin
Heme
Histidine
Degradation
Mutagenesis
Mutation
Biomolecules
Prosthetics
Amino acids
Enzymes
Proteins
Oxidation
Catalysts
Guaiacol
Oxygen
Substrates
Cytochrome-c Peroxidase
Propionates
Horseradish Peroxidase

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Hybridization of Modified-Heme Reconstitution and Distal Histidine Mutation to Functionalize Sperm Whale Myoglobin. / Sato, Hideaki; Hayashi, Takashi; Ando, Tsutomu; Hisaeda, Yoshio; Ueno, Takafumi; Watanabe, Yoshihito.

In: Journal of the American Chemical Society, Vol. 126, No. 2, 21.01.2004, p. 436-437.

Research output: Contribution to journalArticle

Sato, Hideaki ; Hayashi, Takashi ; Ando, Tsutomu ; Hisaeda, Yoshio ; Ueno, Takafumi ; Watanabe, Yoshihito. / Hybridization of Modified-Heme Reconstitution and Distal Histidine Mutation to Functionalize Sperm Whale Myoglobin. In: Journal of the American Chemical Society. 2004 ; Vol. 126, No. 2. pp. 436-437.
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