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Applied and Environmental Microbiology, January 2002, p. 173-180, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.173-180.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of Novosphingobium sp. Strain MT1, a Dominant Polychlorophenol-Degrading Strain in a Groundwater Bioremediation System

Marja A. Tiirola,1 Minna K. Männistö,2 Jaakko A. Puhakka,2 and Markku S. Kulomaa1*

Department of Biological and Environmental Science, University of Jyväskylä, 40351 Jyväskylä,1 Institute of Environmental Engineering and Biotechnology, Tampere University of Technology, 33101 Tampere, Finland2

Received 11 June 2001/ Accepted 9 October 2001

A high-rate fluidized-bed bioreactor has been treating polychlorophenol-contaminated groundwater in southern Finland at 5 to 8°C for over 6 years. We examined the microbial diversity of the bioreactor using three 16S ribosomal DNA (rDNA)-based methods: denaturing gradient gel electrophoresis, length heterogeneity-PCR analysis, and restriction fragment length polymorphism analysis. The molecular study revealed that the process was dependent on a stable bacterial community with low species diversity. The dominant organism, Novosphingobium sp. strain MT1, was isolated and characterized. Novosphingobium sp. strain MT1 degraded the main contaminants of the groundwater, 2,4,6-trichlorophenol, 2,3,4,6-tetrachlorophenol, and pentachlorophenol, at 8°C. The strain carried a homolog of the pcpB gene, coding for the pentachlorophenol-4-monooxygenase in Sphingobium chlorophenolicum. Spontaneous deletion of the pcpB gene homolog resulted in the loss of degradation ability. Phenotypic dimorphism (planktonic and sessile phenotypes), low growth rate (0.14 to 0.15 h–1), and low-copy-number 16S rDNA genes (single copy) were characteristic of strain MT1 and other MT1-like organisms isolated from the bioreactor.

* Corresponding author. Mailing address: Department of Biological and Environmental Science, University of Jyväskylä, P.O. Box 35 (YAB), 40351 Jyväskylä, Finland. Phone: 358-14-260 2273. Fax: 358-14-260 2221. E-mail: kulomaa{at}csc.fi.

Applied and Environmental Microbiology, January 2002, p. 173-180, Vol. 68, No. 1
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.1.173-180.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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