Faculty of Food Science and Engineering, Dunarea de Jos University of Galati, 111 Domneasca Street, 800201 Galati, Romania.
Molecules. 2024 Mar 16;29(6):1327. doi: 10.3390/molecules29061327.
Pepsin, trypsin and proteinase K were used in the present study to hydrolyse the proteins from whole eggs, yolks or whites, and the resulting hydrolysates were characterised in terms of antioxidant and IgE-binding properties, using a combination of in vitro and in silico methods. Based on the degree of hydrolysis (DH) results, the egg yolk proteins are better substrates for all the tested enzymes (DH of 6.2-20.1%) compared to those from egg whites (DH of 2.0-4.4%). The SDS-PAGE analysis indicated that pepsin and proteinase K were more efficient compared to trypsin in breaking the intramolecular peptide bonds of the high molecular weight egg proteins. For all the tested substrates, enzyme-assisted hydrolysis resulted in a significant increase in antioxidant activity, suggesting that many bioactive peptides are encrypted in inactive forms in the parent proteins. The hydrolysates obtained with proteinase K exhibited the highest DPPH radical scavenging activity (124-311 µM Trolox/g protein) and the lowest residual IgE-binding capacity. The bioinformatics tools revealed that proteinase K is able to break the integrity of the main linear IgE-binding epitopes from ovalbumin and ovomucoid. It can be concluded that proteinase K is a promising tool for modulating the intrinsic properties of egg proteins.
本研究使用胃蛋白酶、胰蛋白酶和蛋白酶 K 水解全蛋、蛋黄或蛋清中的蛋白质,并结合使用体外和计算方法来研究水解产物的抗氧化和 IgE 结合特性。根据水解度(DH)的结果,与蛋清蛋白(DH 为 2.0-4.4%)相比,蛋黄蛋白是所有测试酶的更好底物(DH 为 6.2-20.1%)。SDS-PAGE 分析表明,与胰蛋白酶相比,胃蛋白酶和蛋白酶 K 更有效地打破了高分子量鸡蛋蛋白的分子内肽键。对于所有测试的底物,酶辅助水解导致抗氧化活性显著增加,这表明许多生物活性肽以非活性形式存在于母体蛋白中。用蛋白酶 K 获得的水解产物表现出最高的 DPPH 自由基清除活性(124-311 µM Trolox/g 蛋白)和最低的残留 IgE 结合能力。生物信息学工具表明,蛋白酶 K 能够破坏卵清蛋白和卵类黏蛋白中主要线性 IgE 结合表位的完整性。可以得出结论,蛋白酶 K 是一种有前途的调节鸡蛋蛋白固有特性的工具。