Influence of the high pressure on reduction of allergenicity of proteins rMal d1, rDau c1 and orientation quantification of their contents in apple Golden Delicious and in carrot
Abstract
The aim of study was to verify the hypothesis of impact of the high pressure, time and moderate temperature on the reduction of allergenicity of proteins rMal d1 and rDau c1. Recombinant allergens rMal d1 and rDau c1 were treated by high pressures 400, 450, 500, 550 MPa held for 3 and 10 minutes at constant temperature of 25 °C and by the constant pressure of 500 MPa for 10 minutes at temperatures 30, 40 and 50°C.
The basophile activation test was performed using cells from 20 birch allergic patients with oral allergy syndrome to apple and to carrot who have sensitisation to Bet v 1, Mal d 1 and Dau c1. The reactivity of the serum from sensitized patients with treated allergens rMal d1 and rDau c1 at pressure 500MPa for 10 minutes at 30-50°C was verified by Western blot.
The modification of structures rMal d1 and rDau c1 by high pressure was determined by circular dichroism (spectropolarimetr J-810). We did not confirm the influence of high pressure and moderate temperature on allergenicity of rMal d1 and rDau c1 proteins in vitro.
The high pressure of 500 MPa showed major spectral changes in protein rMal d1 without the reduction of its in vitro reactivity. We have attempted to quantify the content of these proteins in apples and carrot.