Association of missense mutations in epoxyalkane coenzyme M transferase with adaptation of Mycobacterium sp. strain JS623 to growth on vinyl chloride

Yang Oh Jin; Samantha Cheung; Nicholas V Coleman; Timothy E Mattes

doi:10.1128/AEM.01320-09

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Association of missense mutations in epoxyalkane coenzyme M transferase with adaptation of Mycobacterium sp. strain JS623 to growth on vinyl chloride

Journal article

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Peer reviewed

Association of missense mutations in epoxyalkane coenzyme M transferase with adaptation of Mycobacterium sp. strain JS623 to growth on vinyl chloride

Yang Oh Jin, Samantha Cheung, Nicholas V Coleman and Timothy E Mattes

Applied and environmental microbiology, Vol.76(11), pp.3413-3419

06/2010

DOI: 10.1128/AEM.01320-09

PMCID: PMC2876445

PMID: 20363787

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Abstract

Vinyl chloride (VC) is a toxic groundwater pollutant associated with plastic manufacture and chlorinated solvent use. Aerobic bacteria that grow on VC as a carbon and energy source can evolve in the laboratory from bacteria that grow on ethene, but the genetic changes involved are unknown. We investigated VC adaptation in two variants (JS623-E and JS623-T) of the ethene-oxidizing Mycobacterium strain JS623. Missense mutations in the EtnE gene developed at two positions (W243 and R257) in cultures exposed to VC but not in cultures maintained on ethene. Epoxyalkane-coenzyme M transferase (EaCoMT) activities in cell extracts of JS623-E and JS623-T (150 and 645 nmol/min/mg protein, respectively) were higher than that of wild-type JS623 (74 nmol/min/mg protein), and in both variant cultures epoxyethane no longer accumulated during growth on ethene. The heterologous expression of two variant etnE alleles (W243G [etnE1] and R257L [etnE2]) from strain JS623 in Mycobacterium smegmatis showed that they had 42 to 59% higher activities than the wild type. Recombinant JS623 cultures containing mutant EtnE genes cloned in the vector pMV261 adapted to growth on VC more rapidly than the wild-type JS623 strain, with incubation times of 60 days (wild type), 1 day (pMVetnE1), and 35 days (pMVetnE2). The JS623(pMVetnE) culture did not adapt to VC after more than 60 days of incubation. Adaptation to VC in strain JS623 is consistently associated with two particular missense mutations in the etnE gene that lead to higher EaCoMT activity. This is the first report to pinpoint a genetic change associated with the transition from cometabolic to growth-linked VC oxidation in bacteria.

Gene Expression

Oxidation-Reduction

Bacterial Proteins - genetics

Molecular Sequence Data

DNA, Bacterial - chemistry

Mutation, Missense

Carbon-Sulfur Lyases - metabolism

Sequence Analysis, DNA

Ethylenes - metabolism

Mycobacterium - growth & development

Mycobacterium - metabolism

Water Pollutants, Chemical - metabolism

DNA, Bacterial - genetics

Carbon-Sulfur Lyases - genetics

Cloning, Molecular

Vinyl Chloride - metabolism

Amino Acid Substitution - genetics

Bacterial Proteins - metabolism

Mycobacterium - enzymology

Details

Title: Subtitle: Association of missense mutations in epoxyalkane coenzyme M transferase with adaptation of Mycobacterium sp. strain JS623 to growth on vinyl chloride
Creators: Yang Oh Jin - Department of Civil and Environmental Engineering, 4105 Seamans Center, University of Iowa, Iowa City, IA 52242, USA
Samantha Cheung
Nicholas V Coleman
Timothy E Mattes
Resource Type: Journal article
Publication Details: Applied and environmental microbiology, Vol.76(11), pp.3413-3419
DOI: 10.1128/AEM.01320-09
PMID: 20363787
PMCID: PMC2876445
NLM abbreviation: Appl Environ Microbiol
ISSN: 1098-5336
eISSN: 1098-5336
Publisher: United States
Language: English
Date published: 06/2010
Academic Unit: Civil and Environmental Engineering
Record Identifier: 9983991989602771

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