Reduction of immunoreactivity of bovine beta-lactoglobulin upon combined physical and proteolytic treatment
| Autor: | Hanne Frøkiær, Patrizia Rasmussen, Pierpaolo Rovere, Francesco Bonomi, Alessandro Fiocchi, Claudio Poiesi, Stefania Iametti, A. Gaiaschi, Patrizia Restani |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Proteolysis
Blotting Western Enzyme-Linked Immunosorbent Assay Lactoglobulins Hydrolysate Enzymatic hydrolysis Settore BIO/10 - Biochimica medicine Chymotrypsin Trypsin Beta-lactoglobulin Chromatography High Pressure Liquid chemistry.chemical_classification Chromatography biology medicine.diagnostic_test Chemistry Hydrolysis Proteolytic enzymes Temperature food and beverages General Medicine Hydrogen-Ion Concentration Enzyme Atmospheric Pressure Biochemistry beta-lactoglobulin high-pressure proteolytic enzymes protein immunoreactivity biology.protein Animal Science and Zoology Electrophoresis Polyacrylamide Gel Settore CHIM/10 - Chimica degli Alimenti Food Science medicine.drug |
| Zdroj: | The Journal of dairy research. 70(1) |
| ISSN: | 0022-0299 |
| Popis: | Bovine beta-lactoglobulin was hydrolyzed with trypsin or chymotrypsin before, during and after treatment at 600 MPa and pH 6.8 for 10 min at 30, 37 and 44 degrees C. The extent of beta-lactoglobulin hydrolysis under pressure was noticeably higher than at atmospheric pressure, particularly when chymotrypsin was used. Addition of proteases at ambient pressure to previously pressure-treated beta-lactoglobulin gave only a modest increase in proteolysis with respect to the untreated protein. Products of enzyme hydrolysis under pressure were separated by reverse-phase HPLC, and were found to be different from those obtained at atmospheric pressure when chymotrypsin was used. The residual immunochemical reactivity of the products of combined pressure-enzyme treatment was assessed on the unresolved hydrolysates by ELISA tests using polyclonal and monoclonal antibodies, and on individual hydrolytic fractions by Western Blotting using sera of paediatric patients allergic to whey proteins in cow milk. The immunoreactivity of the whole hydrolysates was related to their content of residual intact beta-lactoglobulin, and no immunochemical reactivity was found for all the products of chymotrypsin hydrolysis under pressure. The results indicate that chymotrypsin effectively hydrolysed hydrophobic regions of beta-lactoglobulin that were transiently exposed during the pressure treatments and that were not accessible in the native protein or in the protein that had been previously pressure treated. |
| Databáze: | OpenAIRE |
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