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Applied and Environmental Microbiology, January 2006, p. 819-825, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.819-825.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Mycobacterium avium Genes Associated with the Ability To Form a Biofilm

Yoshitaka Yamazaki,1 Lia Danelishvili,1 Martin Wu,1 Molly MacNab,1 and Luiz E. Bermudez1,2*

Department of Biomedical Sciences, College of Veterinary Medicine,1 Department of Microbiology, College of Science, Oregon State University, Corvallis, Oregon 973312

Received 6 September 2005/ Accepted 24 October 2005

Mycobacterium avium is widely distributed in the environment, and it is chiefly found in water and soil. M. avium, as well as Mycobacterium smegmatis, has been recognized to produce a biofilm or biofilm-like structure. We screened an M. avium green fluorescent protein (GFP) promoter library in M. smegmatis for genes involved in biofilm formation on polyvinyl chloride (PVC) plates. Clones associated with increased GFP expression ≥2.0-fold over the baseline were sequenced. Seventeen genes, most encoding proteins of the tricarboxylic acid (TCA) cycle and GDP-mannose and fatty acid biosynthesis, were identified. Their regulation in M. avium was confirmed by examining the expression of a set of genes by real-time PCR after incubation on PVC plates. In addition, screening of 2,000 clones of a transposon mutant bank constructed using M. avium strain A5, a mycobacterial strain with the ability to produce large amounts of biofilm, revealed four mutants with an impaired ability to form biofilm. Genes interrupted by transposons were homologues of M. tuberculosis 6-oxodehydrogenase (sucA), enzymes of the TCA cycle, protein synthetase (pstB), enzymes of glycopeptidolipid (GPL) synthesis, and Rv1565c (a hypothetical membrane protein). In conclusion, it appears that GPL biosynthesis, including the GDP-mannose biosynthesis pathway, is the most important pathway involved in the production of M. avium biofilm.

* Corresponding author. Mailing address: Department of Biomedical Sciences, College of Veterinary Medicine, Oregon State University, 105 Magruder Hall, Corvallis, OR 97331. Phone: (541) 737-6538. Fax: (541) 737-2730. E-mail: luiz.bermudez{at}oregonstate.edu

Applied and Environmental Microbiology, January 2006, p. 819-825, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.819-825.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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