Profiling Mechanisms of Alkane Hydroxylase Activity In Vivo Using the Diagnostic Substrate Norcarane

Elena A. Rozhkova-Novosad, Jong Chan Chae, Gerben J. Zylstra, Erin M. Bertrand, Marselle Alexander-Ozinskas, Dayi Deng, Luke A. Moe, Jan B. van Beilen, Michael Danahy, John Taylor Groves, Rachel N. Austin

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

Mechanistically informative chemical probes are used to characterize the activity of functional alkane hydroxylases in whole cells. Norcarane is a substrate used to reveal the lifetime of radical intermediates formed during alkane oxidation. Results from oxidations of this probe with organisms that contain the two most prevalent medium-chain-length alkane-oxidizing metalloenzymes, alkane ω-monooxygenase (AlkB) and cytochrome P450 (CYP), are reported. The results-radical lifetimes of 1-7 ns for AlkB and less than 100 ps for CYP-indicate that these two classes of enzymes are mechanistically distinguishable and that whole-cell mechanistic assays can identify the active hydroxylase. The oxidation of norcarane by several recently isolated strains (Hydrocarboniphaga effusa AP103, rJ4, and rJ5, whose alkane-oxidizing enzymes have not yet been identified) is also reported. Radical lifetimes of 1-3 ns are observed, consistent with these organisms containing an AlkB-like enzyme and inconsistent with their employing a CYP-like enzyme for growth on hydrocarbons.

Original languageEnglish (US)
Pages (from-to)165-172
Number of pages8
JournalChemistry and Biology
Volume14
Issue number2
DOIs
StatePublished - Feb 1 2007

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

Keywords

  • CHEMBIOL

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