Reduction of immunoreactivity of bovine β-lactoglobulin upon combined physical and proteolytic treatment

Francesco Bonomi, Alessandro Fiocchi, Hanne Frøkiær, Antonella Gaiaschi, Stefania Iametti, Claudio Poiesi, Patrizia Rasmussen, Patrizia Restani, Pierpaolo Rovere

Research output: Contribution to journalArticle

Abstract

Bovine β-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°C. The extent of β-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 β-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 β-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 β-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.

Original languageEnglish
Pages (from-to)51-59
Number of pages9
JournalJournal of Dairy Research
Volume70
Issue number1
DOIs
Publication statusPublished - Feb 2003

Keywords

  • β-lactoglobulin
  • High-pressure
  • Protein immunoreactivity
  • Proteolytic enzymes

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Applied Microbiology and Biotechnology
  • Food Science

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