Journal article
Xylene monooxygenase, a membrane-spanning non-heme diiron enzyme that hydroxylates hydrocarbons via a substrate radical intermediate
Journal of biological inorganic chemistry, v 8(7), pp 733-740
01 Sep 2003
PMID: 12811621
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The non-heme diiron enzyme xylene monooxygenase (XylM) has been shown to hydroxylate hydrocarbons via a hydrogen abstraction-carbon radical recombination mechanism (oxygen rebound). Using the radical clock bicyclo[4.1.0]heptane (norcarane) in a whole-cell assay, and observing the ratio of rearranged 3-(hydroxymethyl)cyclohexene and unrearranged 2-norcaranol products, the lifetime of the substrate radical was determined to be approximately 0.2 ns. The wild-type organism Pseudomonas putida mt-2 and two separate Escherichia coli clones expressing xylMA genes gave similar results. One clone produced the Pseudomonas putida mt-2 XylMA hydroxylase and the other produced Sphingomonas yanoikuyae B1 XylMA hydroxylase. Clones were constructed by inserting genes for xylene monooxygenase and xylene monooxygenase reductase downstream from an IPTG-inducible T7 promoter. Mechanistic investigations using whole-cell assays will facilitate more rapid screening of structure-function relationships and the identification of novel oxygenases. This approach should enable the construction of a picture of the key metalloenzymes and the mechanisms they use in selected parts of the global carbon cycle without requiring the isolation of every protein involved.
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Details
- Title
- Xylene monooxygenase, a membrane-spanning non-heme diiron enzyme that hydroxylates hydrocarbons via a substrate radical intermediate
- Creators
- Rachel N Austin - Bates CollegeKate Buzzi - Bates CollegeEungbin Kim - Yonsei UniversityGerben J Zylstra - Rutgers, The State University of New JerseyJohn T Groves - Princeton UniversityEdward Kim - Computer Science (Computing)
- Publication Details
- Journal of biological inorganic chemistry, v 8(7), pp 733-740
- Publisher
- Springer Nature
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Computer Science
- Web of Science ID
- WOS:000185413100006
- Scopus ID
- 2-s2.0-0141679036
- Other Identifier
- 991019173555504721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology
- Chemistry, Inorganic & Nuclear