Electron-transfer reactions and functionalization of cytochrome P450cam monooxygenase system in reverse micelles

Hirofumi Ichinose, Junji Michizoe, Tatsuo Maruyama, Noriho Kamiya, Masahiro Goto

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Enzyme-based electron-transfer reactions involved in the cytochrome P450 monooxygenase system were investigated in nanostructural reverse micelles. A bacterial flavoprotein, putidaredoxin reductase (PdR), was activated and shown to be capable of catalyzing the electron transport from NADH to electron-carrier proteins such as cytochrome b5 (tCyt-b5) and putidaredoxin (Pdx) in reverse micelles. Ferric tCyt-b5 in reverse micelles was effectively converted to its ferrous form by the exogenous addition of separately prepared reverse micellar solution harboring PdR and NADH. The fact that direct interactions of macromolecular proteins should be possible in the reverse micellar system encouraged us to functionalize a multicomponent monooxygenase system composed of the bacterial cytochrome P450cam (P450cam), putidaredoxin (Pdx), and PdR in reverse micelles. The successful camphor hydroxylation reaction catalyzed by P450cam was significantly dependent on the coexistence of Pdx, PdR, and NADH but not H2O2, suggesting that the oxygen-transfer reactions proceeded via a "monooxygenation" mechanism. This is the first report of a multicomponent cytochrome P450 system exhibiting enzymatic activity in organic media.

Original languageEnglish
Pages (from-to)5564-5568
Number of pages5
JournalLangmuir
Volume20
Issue number13
DOIs
Publication statusPublished - Jun 22 2004

Fingerprint

Camphor 5-Monooxygenase
cytochromes
Micelles
Cytochromes
Mixed Function Oxygenases
micelles
electron transfer
NAD
Proteins
Electrons
Cytochrome P-450 Enzyme System
Camphor
proteins
camphor
Cytochromes b5
Flavoproteins
Hydroxylation
enzymes
Carrier Proteins
electrons

All Science Journal Classification (ASJC) codes

  • Materials Science(all)
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Spectroscopy
  • Electrochemistry

Cite this

Electron-transfer reactions and functionalization of cytochrome P450cam monooxygenase system in reverse micelles. / Ichinose, Hirofumi; Michizoe, Junji; Maruyama, Tatsuo; Kamiya, Noriho; Goto, Masahiro.

In: Langmuir, Vol. 20, No. 13, 22.06.2004, p. 5564-5568.

Research output: Contribution to journalArticle

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