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Appl. Environ. Microbiol., Sep 1996, 3304-3312, Vol 62, No. 9
Copyright © 1996, American Society for Microbiology

Transformation Kinetics of Chlorinated Ethenes by Methylosinus trichosporium OB3b and Detection of Unstable Epoxides by On-Line Gas Chromatography

JET van Hylckama Vlieg, W de Koning and DB Janssen
Department of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, 9747 AG Groningen, The Netherlands

A rapid and accurate method for the determination of transformation kinetics of volatile organic substrates was developed. Concentrations were monitored by on-line gas chromatographic analysis of the headspace of well-mixed incubation mixtures. With this method, the kinetics of transformation of a number of C(inf1) and C(inf2) halogenated alkanes and alkenes by Methylosinus trichosporium OB3b expressing particulate methane monooxygenase or soluble methane monooxygenase (sMMO) were studied. Apparent specific first-order rate constants for cells expressing sMMO decreased in the order of dichloromethane, vinyl chloride, cis-1,2-dichloroethene, trans-1,2-dichloroethene, 1,1-dichloroethene, trichloroethene, chloroform, and 1,2-dichloroethane. During the degradation of trichloroethene, cis-1,2-dichloroethene, trans-1,2-dichloroethene, and vinyl chloride, the formation of the corresponding epoxides was observed. The epoxide of vinyl chloride and the epoxide of trichloroethene, which temporarily accumulated in the medium, were chemically degraded according to first-order kinetics, with half-lives of 78 and 21 s, respectively. Cells expressing sMMO actively degraded the epoxide of cis-1,2-dichloroethene but not the epoxide of trans-1,2-dichloroethene. Methane and acetylene inhibited degradation of the epoxide of cis-1,2-dichloroethene, indicating that sMMO was involved.

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