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Applied and Environmental Microbiology, April 2002, p. 1569-1575, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1569-1575.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Comparison of Extracellular Enzyme Activities and Community Composition of Attached and Free-Living Bacteria in Porous Medium Columns

R. Michael Lehman* and Seán P. O'Connell,{dagger}

Idaho National Engineering and Environmental Laboratory, Biotechnologies Department, Idaho Falls, Idaho 83415-2203

Received 23 October 2001/ Accepted 4 January 2002

Free-living and surface-associated microbial communities in sand-packed columns perfused with groundwater were compared by examination of compositional and functional characteristics. The composition of the microbial communities was assessed by bulk DNA extraction, PCR amplification of 16S ribosomal DNA fragments, separation of these fragments by denaturing gradient gel electrophoresis, and sequence analysis. Community function was assessed by measurement of ß-glucosidase and aminopeptidase extracellular enzyme activities. Free-living populations in the aqueous phase exhibited a greater diversity of phylotypes than populations associated with the solid phase. The attached bacterial community displayed significantly greater ß-glucosidase and aminopeptidase enzyme activities per volume of porous medium than those of the free-living community. On a per-cell basis, the attached community had a significantly higher cell-specific aminopeptidase enzyme activity (1.07 x 10-7 nmol cell-1 h-1) than the free-living community (5.02 x 10-8 nmol cell-1 h-1). Conversely, the free-living community had a significantly higher cell-specific ß-glucosidase activity (1.92 x 10-6 nmol cell-1 h-1) than the surface-associated community (6.08 x 10-7 nmol cell-1 h-1). The compositional and functional differences observed between these two communities may reflect different roles for these distinct but interacting communities in the decomposition of natural organic matter or biodegradation of xenobiotics in aquifers.

* Corresponding author. Mailing address: Idaho National Engineering and Environmental Laboratory, Biotechnologies Department, P.O. Box 1625, Idaho Falls, ID 83415-2203. Phone: (208) 526-3917. Fax: (208) 526-0828. E-mail: mik4{at}inel.gov.

{dagger} Present address: Western Carolina University, Cullowhee, NC 28723.

Applied and Environmental Microbiology, April 2002, p. 1569-1575, Vol. 68, No. 4
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.4.1569-1575.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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