효소에 의한 단백질분해가 유청단백질의 항원성의 저하에 미치는 영향을 조사하기 위한 기본연구로서, 유청단백질의 가수분해특성을 조사하고 competitive inhibition enzyme-linked immunosorbent assay(cELISA)에 의한 항원성의 변화를 검토하였다. 유청단백질의 가수분해는 chymotrypsin, trypsin, pancreatin, 그리고 Aspergillus oryzae유래의 protease를 각기 4시간 동안 행하였다. TNBS(trinitrobenzensulfonic acid)법에 의하여 측정한 유청단백질의 가수분해도(DH)는 chymotrypsin이나 trypsin을 처리한 경우$(5.05{sim}11.47)$보다 Aspergillus oryzae유래의 protease 및 pancreatin을 처리한 경우$(15.67{sim}20.20)$가 훨씬 높게 나타났으며, 각 효소의 처리전에 열처리($75^{circ}C$, 20분)나 pepsin의 처리를 한 경우에 대체로 약간 높게 나타났다. High performance size exclusion chromatography(HPSEC)에 의하여 분자량분포를 조사한 결과, 가수분해물에 따라 10kDa 이상의 polypeptide가 $12{sim}36%$ 정도 존재하였고, 평균분자량은 $4,252{sim}9,132$ dalton, 평균길이는 아미노산 $38{sim}83$개로 나타났다. 또한 쓴맛은 형성되지 않았다. SDS-PAGE의 결과 처리구에 따라 분자량 14.2kDa 이상의 polypeptide가 일부 존재하였으나 native 유청단백질은 대부분 가수분해에 의하여 제거되었음을 확인하였다. 토끼 항WPI항혈청에 의한 cELISA로 검토한 유청단백질 가수분해물의 monovalent 항원성은 효소처리에 의하여 약 $10^{-1.7}{sim}10^{-4.9}$배 또는 그 이하로 저하되었으며 대체로 가수분해가 많이 일어난 분해물은 그 항원성이 낮아지는 것으로 나타났다. 또한 각 처리구내에서는 열 및 pepsin의 전처리후 다음 효소 분해한 유청단백질 가수분해물(CDP, TDP, PDP, ODP)의 경우 그 항원성이 가장 낮았다. 그중에서도 pancreatin 가수분해물(PDP)의 경우 항원성이 거의 상실된 것으로 나타났다.
As a preliminary study about the reduction of the antigenicity of whey protein isolate(WPI) by treatment of chymotrypsin, trypsin, pancreatin, and protease from Aspergillus oryzae, the properties and antigenicities of whey protein hydrolysates(WPH) were investigated. When degrees of hydrolysis (DH) were measured by use of trinitrobenzensulfonic acid(TNBS), the DH of the WPH treated by pancreatin or protease from Aspergillus oryzae$(5.05{sim}11.47)$ were much higher than those of the tryptic or chymotryptic WPH$(15.67{sim}20.20)$. And the pretreatments of heat$(75^{circ}C)$, 20 min and/or pepsin resulted in higher DH of WPH, generally. When the molecular distributions of the WPH were determined by high performance size exclusion chromatography(HPSEC), the ratios of polypeptides with molecular weight more than 10kDa ranged from 12% to 36%, and the average molecular weights and the average peptide lengths of the WPH were $4,252{sim}9,132$ dalton and $38{sim}83$ amino acids, respectively. And there was no bitter taste in all of the WPH. Results of SDS-PAGE showed that most of intact native proteins were eliminated by the enzymatic hydrolysis but there were a few bands of peptides larger than 14.2 kDa in some WPH. When antigenicity was assayed by competitive inhibition enzyme-linked immunosorbent assay(cELISA), monovalent antigenicity of WPH to rabbit anti-WPI antiserum were lowered to $10^{-1.7}-10^{-4.9}$ times and less by the enzymatic hydrolysis. And the pretreatments of heat and pepsin resulted in the lowest antigenicicy within a group of enzymatic hydrolysis, especially in case of the pancreatic hydrolysate(PDP) whose antigenicity was found almost to be removed.
