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Applied and Environmental Microbiology, May 2000, p. 1834-1843, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Isolation of Adherent Polycyclic Aromatic Hydrocarbon (PAH)-Degrading Bacteria Using PAH-Sorbing Carriers

Leen Bastiaens,1,2 Dirk Springael,1,* Pierre Wattiau,3 Hauke Harms,4 Rupert deWachter,5 Hubert Verachtert,2 and Ludo Diels1

Environmental Technology, Vlaamse Instelling voor Technologisch Onderzoek, B-2400 Mol,1 Laboratory of Industrial Microbiology and Biochemistry, Katholieke Universiteit Leuven B-3001 Leuven,2 Microbial Pathogenesis Unit, Université Catholique de Louvain, B-1200 Brussels,3 and Department of Biochemistry, University of Antwerp B-2610 Antwerp,5 Belgium, and Swiss Federal Institute for Environmental Science and Technology CH-8600 Dübendorf, Switzerland4

Received 24 September 1999/Accepted 26 January 2000

Two different procedures were compared to isolate polycyclic aromatic hydrocarbon (PAH)-utilizing bacteria from PAH-contaminated soil and sludge samples, i.e., (i) shaken enrichment cultures in liquid mineral medium in which PAHs were supplied as crystals and (ii) a new method in which PAH degraders were enriched on and recovered from hydrophobic membranes containing sorbed PAHs. Both techniques were successful, but selected from the same source different bacterial strains able to grow on PAHs as the sole source of carbon and energy. The liquid enrichment mainly selected for Sphingomonas spp., whereas the membrane method exclusively led to the selection of Mycobacterium spp. Furthermore, in separate membrane enrichment set-ups with different membrane types, three repetitive extragenic palindromic PCR-related Mycobacterium strains were recovered. The new Mycobacterium isolates were strongly hydrophobic and displayed the capacity to adhere strongly to different surfaces. One strain, Mycobacterium sp. LB501T, displayed an unusual combination of high adhesion efficiency and an extremely high negative charge. This strain may represent a new bacterial species as suggested by 16S rRNA gene sequence analysis. These results indicate that the provision of hydrophobic sorbents containing sorbed PAHs in the enrichment procedure discriminated in favor of certain bacterial characteristics. The new isolation method is appropriate to select for adherent PAH-degrading bacteria, which might be useful to biodegrade sorbed PAHs in soils and sludge.

* Corresponding author. Mailing address: Vito, Boeretang 200, B-2400 Mol, Belgium. Phone: 32 14 33 51 76. Fax: 32 14 58 05 23. E-mail: dirk.springael{at}vito.be.

Applied and Environmental Microbiology, May 2000, p. 1834-1843, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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