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Applied and Environmental Microbiology, October 2004, p. 6098-6112, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.6098-6112.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Transcriptional Analysis of Biofilm Formation Processes in the Anaerobic, Hyperthermophilic Bacterium Thermotoga maritima

Marybeth A. Pysz,{dagger} Shannon B. Conners, Clemente I. Montero, Keith R. Shockley, Matthew R. Johnson, Donald E. Ward,{ddagger} and Robert M. Kelly*

Department of Chemical Engineering, North Carolina State University, Raleigh, North Carolina

Received 24 February 2004/ Accepted 2 April 2004

Thermotoga maritima, a fermentative, anaerobic, hyperthermophilic bacterium, was found to attach to bioreactor glass walls, nylon mesh, and polycarbonate filters during chemostat cultivation on maltose-based media at 80°C. A whole-genome cDNA microarray was used to examine differential expression patterns between biofilm and planktonic populations. Mixed-model statistical analysis revealed differential expression (twofold or more) of 114 open reading frames in sessile cells (6% of the genome), over a third of which were initially annotated as hypothetical proteins in the T. maritima genome. Among the previously annotated genes in the T. maritima genome, which showed expression changes during biofilm growth, were several that corresponded to biofilm formation genes identified in mesophilic bacteria (i.e., Pseudomonas species, Escherichia coli, and Staphylococcus epidermidis). Most notably, T. maritima biofilm-bound cells exhibited increased transcription of genes involved in iron and sulfur transport, as well as in biosynthesis of cysteine, thiamine, NAD, and isoprenoid side chains of quinones. These findings were all consistent with the up-regulation of iron-sulfur cluster assembly and repair functions in biofilm cells. Significant up-regulation of several ß-specific glycosidases was also noted in biofilm cells, despite the fact that maltose was the primary carbon source fed to the chemostat. The reasons for increased ß-glycosidase levels are unclear but are likely related to the processing of biofilm-based polysaccharides. In addition to revealing insights into the phenotype of sessile T. maritima communities, the methodology developed here can be extended to study other anaerobic biofilm formation processes as well as to examine aspects of microbial ecology in hydrothermal environments.

* Corresponding author. Mailing address: Department of Chemical Engineering, North Carolina State University, Raleigh, NC 27695-7905. Phone: (919) 515-6396. Fax: (919) 515-3465. E-mail: rmkelly{at}eos.ncsu.edu.

{dagger} Present address: Roswell Park Cancer Institute, Department of Pharmacology and Therapeutic, Buffalo, NY 14263.

{ddagger} Present address: Genencor International, Inc., Palo Alto, CA 94304.

Applied and Environmental Microbiology, October 2004, p. 6098-6112, Vol. 70, No. 10
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.10.6098-6112.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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