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Applied and Environmental Microbiology, July 2000, p. 2703-2710, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Effect of Model Sorptive Phases on Phenanthrene Biodegradation: Molecular Analysis of Enrichments and Isolates Suggests Selection Based on Bioavailability

M. Friedrich,1,* R. J. Grosser,1,dagger E. A. Kern,2 W. P. Inskeep,1,2 and D. M. Ward1

Department of Land Resources and Environmental Sciences1 and Department of Microbiology and Center for Biofilm Engineering,2 Montana State University, Bozeman, Montana 59717

Received 20 October 1999/Accepted 31 March 2000

Reduced bioavailability of nonpolar contaminants due to sorption to natural organic matter is an important factor controlling biodegradation of pollutants in the environment. We established enrichment cultures in which solid organic phases were used to reduce phenanthrene bioavailability to different degrees (R. J. Grosser, M. Friedrich, D. M. Ward, and W. P. Inskeep, Appl. Environ. Microbiol. 66:2695-2702, 2000). Bacteria enriched and isolated from contaminated soils under these conditions were analyzed by denaturing gradient gel electrophoresis (DGGE) and sequencing of PCR-amplified 16S ribosomal DNA segments. Compared to DGGE patterns obtained with enrichment cultures containing sand or no sorptive solid phase, different DGGE patterns were obtained with enrichment cultures containing phenanthrene sorbed to beads of Amberlite IRC-50 (AMB), a weak cation-exchange resin, and especially Biobead SM7 (SM7), a polyacrylic resin that sorbed phenanthrene more strongly. SM7 enrichments selected for mycobacterial phenanthrene mineralizers, whereas AMB enrichments selected for a Burkholderia sp. that degrades phenanthrene. Identical mycobacterial and Burkholderia 16S rRNA sequence segments were found in SM7 and AMB enrichment cultures inoculated with contaminated soil from two geographically distant sites. Other closely related Burkholderia sp. populations, some of which utilized phenanthrene, were detected in sand and control enrichment cultures. Our results are consistent with the hypothesis that different phenanthrene-utilizing bacteria inhabiting the same soils may be adapted to different phenanthrene bioavailabilities.

* Corresponding author. Present address: Max Planck Institute for Terrestrial Microbiology, Karl-von-Frisch-Straße, D-35043 Marburg/Lahn, Germany. Phone: 49-6421-178830. Fax: 49-6421-178809. E-mail: friedric{at}mailer.uni-marburg.de.

dagger Present address: NRMRL, US EPA, Cincinnati, OH 45268.

Applied and Environmental Microbiology, July 2000, p. 2703-2710, Vol. 66, No. 7
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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