Characterization of a Potential Listeria monocytogenes Virulence Factor Associated with Attachment to Fresh Produce

Abstract

ABSTRACT A study to determine the attachment of L. monocytogenes serotype 4b strain F2365 on vegetables and fruits was conducted. In an initial study, we screened 32 genes encoding surface proteins and lipases of the strain to find highly expressed genes on lettuce leaves. The results showed that transcription levels of LMOf2365_0413, LMOf2365_0498, LMOf2365_0859, LMOf2365_2052, and LMOf2365_2812 were significantly upregulated on lettuce leaves. In silico analysis showed that LMOf2365_0859 contains a putative cellulose binding domain. Thus, we hypothesized that this gene may be involved in an attachment to vegetables, and named it lcp (gene encoding Listeria cellulose binding protein [LCP]). lcp mutant (Δ lcp ) and lcp complement (F2365::pMAD:: cat :: lcp ) strains were generated by homologous recombination. The abilities of a wild-type (WT) strain, the Δ lcp strain, and the complemented strain to attach to lettuce leaves were evaluated, which indicated that the attachment of the Δ lcp strain to lettuce was significantly less than that of the WT and the complemented strains. Similar results were observed for baby spinach and cantaloupe. Fluorescence microscopy and field emission scanning microscopy analysis further supported these findings. The binding of L. monocytogenes to cellulose was determined using cellulose acetate-coated plates. The results showed that a binding ability of the Δ lcp strain was significantly lower than that of the wild type. Combined, these results strongly suggest that LCP plays an important role in an attachment to vegetables and fruits.