Effects of heat treatment and pectin addition on beta-lactoglobulin allergenicity.
The specific effects of heat treatment and/or addition of low/high-methylated pectin (LMP/HMP) on the allergenicity of beta-lactoglobulin (beta-Lg) and its hydrolysis products were investigated through a two-step in vitro digestion approach. beta-Lg was first hydrolyzed by pepsin and then by a trypsin/chymotrypsin (T/C) mixture done in a dialysis bag with a molecular weight cutoff of 1000. The protein digestion was followed by SDS-PAGE electrophoresis performed on each digestion product, and their in vitro allergenicity was analyzed by immunoblotting. Such procedure was applied on beta-Lg samples mixed with the two kinds of pectin before or after heating (80 degrees C, 25 min) to determine the respective impact of heat treatment and pectin addition. Heat denaturation improved significantly the susceptibility of beta-Lg against the pepsin and the T/C. This effect, which was coupled to a reduction in immunoreactivity of the digested beta-Lg, appeared to be distinctively modulated by LMP and HMP. Through nonspecific interaction with the beta-Lg, pectin could reduce the accessibility of cleavage sites and/or epitope sequences. This mechanism of action is discussed in relation to the intra- and intermolecular interactions between beta-Lg and pectin initiated under the experimental conditions.
Peyron
Stéphane
S
Unité Mixte de Recherche Ingénierie des Agropolymères et des Technologies Emergentes, Université Montpellier II, cc023, Pl. E Bataillon, 34095 Montpellier, France.
Mouécoucou
Justine
J
Frémont
Sophie
S
Sanchez
Christian
C
Gontard
Nathalie
N
D000485:Allergens / Q000276:immunology*; D002918:Chymotrypsin / Q000378:metabolism; D005512:Food Hypersensitivity; D006358:Hot Temperature; D006868:Hydrolysis; D007782:Lactoglobulins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D008745:Methylation; D010316:Particle Size; D010368:Pectins / Q000737:chemistry / Q000494:pharmacology*; D010434:Pepsin A / Q000378:metabolism; D011489:Protein Denaturation; D014357:Trypsin / Q000378:metabolism
D000485:Allergens; D007782:Lactoglobulins; D010368:Pectins; D002918:Chymotrypsin; D014357:Trypsin; D010434:Pepsin A