Abstract
Wheat bran (WB) from Triticum aestivum has many beneficial effects on human health. To the best of our knowledge, very little has been published about its ability to prevent pathogenic bacterial adhesion in the intestine. Here, a WB extract was fractionated using different strategies, and the obtained fractions were tested in different in vitro methodologies to evaluate their interference in the attachment of enterotoxigenic Escherichia coli (ETEC) K88 to intestinal porcine epithelial cells (IPEC-J2) with the aim of identifying the putative anti-adhesive molecules. It was found that a proteinaceous compound in the >300-kDa fraction mediates the recognition of ETEC K88 to IPEC-J2. Further fractionation of the >300-kDa sample by size-exclusion chromatography showed several proteins below 90 kDa, suggesting that the target protein belongs to a high-molecular-weight (MW) multi-component protein complex. The identification of some relevant excised bands was performed by mass spectrometry (MS) and mostly revealed the presence of various protease inhibitors (PIs) of low MW: Serpin-Z2B, Class II chitinase, endogenous alpha-amylase/subtilisin inhibitor and alpha-amylase/trypsin inhibitor CM3. Furthermore, an incubation of the WB extract with ETEC K88 allowed for the identification of a 7S storage protein globulin of wheat, Globulin 3 of 66 kDa, which may be one of the most firmly attached WB proteins to ETEC K88 cells. Further studies should be performed to gain an understanding of the molecular recognition of the blocking process that takes place. All gathered information can eventually pave the way for the development of novel anti-adhesion therapeutic agents to prevent bacterial pathogenesis.
Highlights
The possibility that pathogens can be inhibited by naturally occurring anti-adhesive compounds is especially attractive and has captured significant attention [1]
Nature of the putative compound of Wheat bran (WB) involved in enterotoxigenic E. coli (ETEC) K88 adhesion
Since anti-adhesives based on carbohydrate structures are more common than are those based on proteins [22,23], a carbohydrate digestion was carried out by using both O-glycosidase and neuraminidase in order to detect carbohydrate participation in the ETEC K88 binding
Summary
The possibility that pathogens can be inhibited by naturally occurring anti-adhesive compounds is especially attractive and has captured significant attention [1]. Plant lectins are well represented in human and animal diets and many of these are well characterized [3], their application to anti-adhesion therapies is a recent strategy that has become an alternative to antibiotics. These lectins could interact with host-cell receptors to block adhesion by competition. They could interact with bacterial adhesins to enhance the clearance of bacteria by exclusion [4]. Numerous investigations involving plant extracts have recently been performed to reduce bacterial adhesion [5,6,7]
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