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Applied and Environmental Microbiology, July 2000, p. 2743-2747, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Anaerobic Naphthalene Degradation by a Sulfate-Reducing Enrichment Culture†

Rainer U. Meckenstock,1,* Eva Annweiler,2 Walter Michaelis,2 Hans H. Richnow,2,Dagger and Bernhard Schink1

Department of Biology, University of Konstanz, D-78457 Konstanz,1 and Institute of Biogeochemistry and Marine Chemistry, University of Hamburg, D-20146 Hamburg,2 Germany

Received 23 December 1999/Accepted 11 April 2000

Anaerobic naphthalene degradation by a sulfate-reducing enrichment culture was studied by substrate utilization tests and identification of metabolites by gas chromatography-mass spectrometry. In substrate utilization tests, the culture was able to oxidize naphthalene, 2-methylnaphthalene, 1- and 2-naphthoic acids, phenylacetic acid, benzoic acid, cyclohexanecarboxylic acid, and cyclohex-1-ene-carboxylic acid with sulfate as the electron acceptor. Neither hydroxylated 1- or 2-naphthoic acid derivatives and 1- or 2-naphthol nor the monoaromatic compounds ortho-phthalic acid, 2-carboxy-1-phenylacetic acid, and salicylic acid were utilized by the culture within 100 days. 2-Naphthoic acid accumulated in all naphthalene-grown cultures. Reduced 2-naphthoic acid derivatives could be identified by comparison of mass spectra and coelution with commercial reference compounds such as 1,2,3,4-tetrahydro-2-naphthoic acid and chemically synthesized decahydro-2-naphthoic acid. 5,6,7,8-Tetrahydro-2-naphthoic acid and octahydro-2-naphthoic acid were tentatively identified by their mass spectra. The metabolites identified suggest a stepwise reduction of the aromatic ring system before ring cleavage. In degradation experiments with [1-13C]naphthalene or deuterated D8-naphthalene, all metabolites mentioned derived from the introduced labeled naphthalene. When a [13C]bicarbonate-buffered growth medium was used in conjunction with unlabeled naphthalene, 13C incorporation into the carboxylic group of 2-naphthoic acid was shown, indicating that activation of naphthalene by carboxylation was the initial degradation step. No ring fission products were identified.

* Corresponding author. Mailing address: Department of Biology, University of Konstanz, Universitätstr. 10, D-78457 Konstanz, Germany. Phone: 49-7531-884541. Fax: 49-7531-882966. E-mail: rainer.meckenstock{at}uni-konstanz.de.

dagger Publication 88 of Deutsche Forschungsgemeinschaft priority program 546, Geochemical Processes with Long-Term Effects in Anthropogenically Affected Seepage and Groundwater.

Dagger Present address: UFZ Leipzig-Halle GmbH, D-04318 Leipzig, Germany.

Applied and Environmental Microbiology, July 2000, p. 2743-2747, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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