물리적 처리에 의한 porcine serum albumin의 allergenicity 변화

Abstract

Ci-ELISA, SDS-PAGE, Immunoblotting을 이용하여 여러 가지 물리적 처리에 의한 돼지고기의 allergen 및 allergenicity의 변화를 알아본 결과는 다음과 같다. 1. P-IgG에 대한 돼지고기의 주 allergen인 PSA와 pork meat extract의 allergenicity 변화 결과는 다음과 같다. 1-1. 가열처리, 가압가열처리, 방사선 처리된 PSA의 allergenicity 변 화 결과 100℃, 20 min 가열 처리시 52%, 가압 가열(121℃, 1 kg/cm²) 30 min 처리시 29%, 3 kGy의 방사선 조사시 30% 이하로 결합력이 크게 감소하였다. Immunoblotting에서도 PSA band는 항체 와 거의 반응하지 않았다. 1-2. 가열처리된 pork meat extract의 allergenicity 변화 결과, 100℃, 121℃에서 처리시 PSA가 p-IgG와의 결합력이 23%, 6%로 크게 감소 되었으며 immunoblotting 결과에서도 반응하지 않았다. 1-3. 초고압 처리에 의한 PSA의 allergenicity 변화를 알아본 결과 400 MPa에서도 PSA와 p-IgG와의 결합력이 높게 유지되었으며, 또 한 초음파 처리시에도 77%정도로 높게 유지되었다. 1-4. 2,450 MHz의 microwave 처리와 5, 10, 30, 60 min간 초음파 처 리하여 allergenicity 변화를 알아본 결과 20 min간 열을 배제하고 microwave 처리시 100% 정도의 높은 결합력을 보였으나 10 min간 중탕으로 microwave 처리에 의해서는 31%의 낮은 결합력을 보였다. 2. Pig-allergic patients' serum에 대한 돼지고기의 주 allergen인 PSA의 allergenicity 변화 결과는 121℃, 30 min 가압가열에 의해 10%, 3 kGy 방사선 조사에 의해 15% 정도의 낮은 결합력을 보였으며 immunoblotting 결과에서도 환자혈청과 반응하지 않았다. 이상의 결과에서 주 allergen인 PSA는 100℃ 이상의 가열처리, 가압가열처리, 방사선 조사에 의해 allergenicity가 크게 감소하였으며, 이는 저 알레르기 돼기고기 식품 제조에 응용할 수 있을 것으로 보인다.

The aim of this study was to evaluate the effects of physical treatment(heating, autoclaving, high hydrostatic pressure, sonication, microwave and gamma irradiation) on the allergenicity of porcine serum albumin(PSA) and pork meat(PM). This study was performed using a competitive indirect immunosorbent assay(Ci-ELISA), sodium dodesyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and immunoblotting to investigate the change in the binding ability of PSA and PM altered by phsical treatment to p-IgG and the serum of patients who were allergic to pigs. The following changes were observed in the allergenicity of PSA and PM to p-IgG. The heat treatment at 100℃ for 20 min on PSA and PM was able to significantly decrease the binding ability of PSA to p-IgG at 52% and 23%, respectively. However heat treatment at 60℃ and 80℃ was not able to decrease the binding ability of PSA to p-IgG. An autoclave treatment on PSA for 30 min and PM for 15 min decreased the binding ability to p-IgG at 29% and 6%, respectively. The binding ability of 3 kGy gamma-irradiated PSA was markedly decreased to 30%. SDS-PAGE and immunoblotting of heat-treated PSA and PM over 100℃ and gamma-irradiated PSA revealed that the intensity of the 66 kD band decreased and that p-IgG barely recognized their presence. The microwave treatment of PSA was able to decrease the binding ability depending on the duration of the treatment. Notably, the binding ability of PSA treated for 10 min was 29%. Immunoblotting of the microwave-treated PSA revealed that the p-IgG was hardly recognized in 5 and 10 min treatments. Exposure to high hydrostatic pressure, sonication, and microwave without heat did not affect the allergen and allergenicity of PSA. The following changes were observed in the allergenicity of the serum of patients' with pig allergies. The binding ability of the autoclave-treated and gamma-irradiated PSA decreased more in the serum of the patients with pig allergies than in p-IgG. Immunoblotting revealed that the serum of the patients with pig allergies hardly even recognized the PSA.