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Applied and Environmental Microbiology, July 2005, p. 3797-3805, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3797-3805.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Isolation of Soil Bacteria Adapted To Degrade Humic Acid-Sorbed Phenanthrene

D. J. Vacca,¹ W. F. Bleam,^1,2 and W. J. Hickey^1,2*

Department of Soil Science,¹ Molecular and Environmental Toxicology Center, University of Wisconsin—Madison, Madison, Wisconsin²

Received 1 November 2004/ Accepted 5 January 2005

The goal of these studies was to determine how sorption by humic acids affected the bioavailability of polynuclear aromatic hydrocarbons (PAHs) to PAH-degrading microbes. Micellar solutions of humic acid were used as sorbents, and phenanthrene was used as a model PAH. Enrichments from PAH-contaminated soils established with nonsorbed phenanthrene yielded a total of 25 different isolates representing a diversity of bacterial phylotypes. In contrast, only three strains of Burkholderia spp. and one strain each of Delftia sp. and Sphingomonas sp. were isolated from enrichments with humic acid-sorbed phenanthrene (HASP). Using [¹⁴C]phenanthrene as a radiotracer, we verified that only HASP isolates were capable of mineralizing HASP, a phenotype hence termed "competence." Competence was an all-or-nothing phenotype: noncompetent strains showed no detectable phenanthrene mineralization in HASP cultures, but levels of phenanthrene mineralization effected by competent strains in HASP and NSP cultures were not significantly different. Levels and rates of phenanthrene mineralization exceeded those predicted to be supported solely by the metabolism of phenanthrene in the aqueous phase of HASP cultures. Thus, competent strains were able to directly access phenanthrene sorbed by the humic acids and did not rely on desorption for substrate uptake. To the best of our knowledge, this is the first report of (i) a selective interaction between aerobic bacteria and humic acid molecules and (ii) differential bioavailability to bacteria of PAHs sorbed to a natural biogeopolymer.

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* Corresponding author. Mailing address: Department of Soil Science, University of Wisconsin—Madison, Madison, WI 53706-1299. Phone: (608) 262-9018. Fax: (608) 265-2595. E-mail: wjhickey{at}wisc.edu.

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Applied and Environmental Microbiology, July 2005, p. 3797-3805, Vol. 71, No. 7
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.7.3797-3805.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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