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Applied and Environmental Microbiology, April 2008, p. 2384-2390, Vol. 74, No. 8
0099-2240/08/$08.00+0     doi:10.1128/AEM.01854-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Polysaccharides Cellulose, Poly-β-1,6-N-Acetyl-D-Glucosamine, and Colanic Acid Are Required for Optimal Binding of Escherichia coli O157:H7 Strains to Alfalfa Sprouts and K-12 Strains to Plastic but Not for Binding to Epithelial Cells{triangledown}

Ann G. Matthysse,1* Rajendar Deora,2 Meenu Mishra,2 and Alfredo G. Torres3

Department of Biology, University of North Carolina, Chapel Hill, North Carolina 27599-3280,1 Department of Microbiology and Immunology, Wake Forest University Health Sciences, Medical Center Blvd., Room 5086, Winston-Salem, North Carolina 27157,2 Department of Microbiology and Immunology, Department of Pathology, and Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas 77555-10703

Received 9 August 2007/ Accepted 14 February 2008

When Escherichia coli O157:H7 bacteria are added to alfalfa sprouts growing in water, the bacteria bind tightly to the sprouts. In contrast, laboratory K-12 strains of E. coli do not bind to sprouts under similar conditions. The roles of E. coli O157:H7 lipopolysaccharide (LPS), capsular polysaccharide, and exopolysaccharides in binding to sprouts were examined. An LPS mutant had no effect on the binding of the pathogenic strain. Cellulose synthase mutants showed a significant reduction in binding; colanic acid mutants were more severely reduced, and binding by poly-β-1,6-N-acetylglucosamine (PGA) mutants was barely detectable. The addition of a plasmid carrying a cellulose synthase gene to K-12 strains allowed them to bind to sprouts. A plasmid carrying the Bps biosynthesis genes had only a marginal effect on the binding of K-12 bacteria. However, the introduction of the same plasmid allowed Sinorhizobium meliloti and a nonbinding mutant of Agrobacterium tumefaciens to bind to tomato root segments. These results suggest that although multiple redundant protein adhesins are involved in the binding of E. coli O157:H7 to sprouts, the polysaccharides required for binding are not redundant and each polysaccharide may play a distinct role. PGA, colanic acid, and cellulose were also required for biofilm formation by a K-12 strain on plastic, but not for the binding of E. coli O157:H7 to mammalian cells.

* Corresponding author. Mailing address: Department of Biology, Coker Hall, University of North Carolina, Chapel Hill, NC 27599-3280. Phone: (919) 962-6941. Fax: (919) 962-1625. E-mail: ann_matthysse{at}unc.edu

{triangledown} Published ahead of print on 29 February 2008.

Applied and Environmental Microbiology, April 2008, p. 2384-2390, Vol. 74, No. 8
0099-2240/08/$08.00+0     doi:10.1128/AEM.01854-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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