Identification of proteolytic enzymes from Eriphia verrucosa and Palinurus elephas capable to degrade gliadin

In small intestinal disease, coeliac sprue, proline-rich gluten peptides from wheat, rye and barley are relatively resistant to gastrointestinal digestion, and therefore remain in the intestinal lumen to elicit immunopathology in genetically susceptible individuals. Since most serine endopeptidases are unable to hydrolyse proline residues, proline specific proteases may be therapeutic keys in digestive diseases. Partial hydrolysis reduces the risk of allergenic sensitization while total hydrolysis ensures the elimination of the allergenicity of whey protein (Villad´oniga and others 2007). Kimoto and others (1998) reported that 18-, 31-, 37- and 58-kDa wheat allergens were recognized by the IgE antibodies in the sera of 20%, 60%, 60%, and 57% of allergic patients, respectively. Gliadin has been considered to cause wheat-dependent exercise-induced anaphylaxis. In this work, we used a new mixture of extracted marine proteases to digest gliadin. The enzymes were extracted from epatopancreas of Eriphia verrucosa and Palinurus elephas; the tissues were homogenised and after differential centrifugation the enzymes were used in enzymatic assay. The digestion was performed at low temperature (4°C). After digestions the substrate was analysed in SDS-PAGE and the allergenic fragments result are completely digested by the action of both different marine species enzymes. At present, we are try to identify the marine prolyl endopeptidases and/or prolyl oligopeptidases, families of serine proteases having the ability to hydrolyse the peptide bond on the carboxyl side of a proline residue. These proline specific enzymes are widely distributed in bacteria, fungi, animals and plants.