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Journal of Bacteriology, September 2005, p. 6128-6136, Vol. 187, No. 17
0021-9193/05/$08.00+0     doi:10.1128/JB.187.17.6128-6136.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Biodiversity-Based Identification and Functional Characterization of the Mannose-Specific Adhesin of Lactobacillus plantarum

Gabriele Pretzer,1,2 Johannes Snel,1 Douwe Molenaar,1,3 Anne Wiersma,1,3 Peter A. Bron,1,3,{dagger} Jolanda Lambert,1,3 Willem M. de Vos,3 Roelof van der Meer,1,3 Mari A. Smits,2 and Michiel Kleerebezem1,3*

NIZO Food Research, P.O. Box 20, 6710 BA Ede, The Netherlands,1 Animal Sciences Group of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands,2 Wageningen Centre for Food Sciences, P.O. Box 557, 6700 AN Wageningen, The Netherlands3

Received 4 March 2005/ Accepted 16 June 2005

Lactobacillus plantarum is a frequently encountered inhabitant of the human intestinal tract, and some strains are marketed as probiotics. Their ability to adhere to mannose residues is a potentially interesting characteristic with regard to proposed probiotic features such as colonization of the intestinal surface and competitive exclusion of pathogens. In this study, the variable capacity of 14 L. plantarum strains to agglutinate Saccharomyces cerevisiae in a mannose-specific manner was determined and subsequently correlated with an L. plantarum WCFS1-based genome-wide genotype database. This led to the identification of four candidate mannose adhesin-encoding genes. Two genes primarily predicted to code for sortase-dependent cell surface proteins displayed a complete gene-trait match. Their involvement in mannose adhesion was corroborated by the finding that a sortase (srtA) mutant of L. plantarum WCFS1 lost the capacity to agglutinate S. cerevisiae. The postulated role of these two candidate genes was investigated by gene-specific deletion and overexpression in L. plantarum WCFS1. Subsequent evaluation of the mannose adhesion capacity of the resulting mutant strains showed that inactivation of one candidate gene (lp_0373) did not affect mannose adhesion properties. In contrast, deletion of the other gene (lp_1229) resulted in a complete loss of yeast agglutination ability, while its overexpression quantitatively enhanced this phenotype. Therefore, this gene was designated to encode the mannose-specific adhesin (Msa; gene name, msa) of L. plantarum. Domain homology analysis of the predicted 1,000-residue Msa protein identified known carbohydrate-binding domains, further supporting its role as a mannose adhesin that is likely to be involved in the interaction of L. plantarum with its host in the intestinal tract.

* Corresponding author. Mailing address: NIZO Food Research, P.O. Box 20, 6710 BA Ede, The Netherlands. Phone: 31-318-659629. Fax: 31-318-650400. E-mail: Michiel.Kleerebezem{at}nizo.nl.

{dagger} Present address: APC (UCC), Cork, Ireland.

Journal of Bacteriology, September 2005, p. 6128-6136, Vol. 187, No. 17
0021-9193/05/$08.00+0     doi:10.1128/JB.187.17.6128-6136.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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