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Applied and Environmental Microbiology, June 2005, p. 2902-2909, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.2902-2909.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of a Gene Cluster for the Formation of Extracellular Polysaccharide Precursors in the Chemolithoautotroph Acidithiobacillus ferrooxidans

Marlen Barreto,1,2 Eugenia Jedlicki,2 and David S. Holmes1*

Laboratory of Bioinformatics and Genome Biology, Andrés Bello University, and Millennium Institute of Fundamental and Applied Biology,1 Program of Cellular & Molecular Biology, University of Chile, Santiago, Chile2

Received 3 November 2004/ Accepted 21 December 2004

A cluster of five genes, proposed to be involved in the formation of extracellular polysaccharide (EPS) precursors via the Leloir pathway, have been identified in the acidophilic autotroph Acidithiobacillus ferrooxidans. The order of the genes is luxA-galE-galK-pgm-galM, encoding a LuxA-like protein, UDP-glucose 4-epimerase, galactokinase, phosphoglucomutase, and galactose mutarotase, respectively. The gal cluster forms a single transcriptional unit and is therefore an operon. Two other putative genes of the Leloir pathway, galU, potentially encoding UDP-glucose pyrophosphorylase, and a gene designated galT-like, which may encode a galactose-1-phosphate uridylyltransferase-like activity, were found unlinked in the genome. Using semiquantitative reverse transcription-PCR, the genes of the gal operon were shown to be expressed more during growth in iron medium than in growth in sulfur medium. The functions of galE, pgm, galU, and the galT-like gene were validated by complementation of Escherichia coli mutants and by in vitro enzyme assays. The data suggest that A. ferrooxidans is capable of synthesizing the EPS precursors UDP-glucose and UDP-galactose. In addition, genes rfbA, -B, -C, and -D were identified in the genome of A. ferrooxidans, suggesting that it can also synthesize the EPS precursor dTDP-rhamnose. Since EPSs constitute the major bulk of biofilms, this study may provide an initial model for the metabolic pathways involved in biofilm formation in A. ferrooxidans and aid in understanding the role of biofilms in mineral leaching and the formation of acid mine drainage.

* Corresponding author. Mailing address: Laboratory of Bioinformatics and Genome Biology, Andres Bello University and Millennium Institute of Fundamental and Applied Biology, Santiago, Chile. Phone: 56-2-239-8969. Fax: 56-2-237-2259. E-mail: dsholmes2000{at}yahoo.com.

Applied and Environmental Microbiology, June 2005, p. 2902-2909, Vol. 71, No. 6
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.6.2902-2909.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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