从蛋黄可溶性蛋白中提取抗氧化活性肽的研究

H. Ibrahim, Ai Hitotsumatsu, T. Miyata
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引用次数: 0

摘要

背景:鸡蛋是一种营养丰富的食物,含有蛋白质、脂肪、碳水化合物、矿物质和维生素。虽然蛋黄中含有许多蛋白质,但很少有关于它们对健康有益的研究。目的:研究从蛋黄可溶性蛋白中提取的抗氧化肽。在酸性条件下用水稀释后,用活性炭去除脂质和脂蛋白。材料和方法:与蛋白中含有3.6克蛋白质相比,单个大鸡蛋中蛋黄含有2.7克蛋白质。将蛋黄可溶性蛋白(EYsP)在pH 3.0条件下用胃蛋白酶(P-EYsP)消化2h,然后用胰蛋白酶(P-EYsP- t)、α-凝乳胰蛋白酶(P-EYsP-α)或两种酶(P-EYsP- t /α)消化。采用sepphacryl S-100凝胶过滤对活性消解物进行分离,并采用反相高效液相色谱法对活性消解物进行分离。利用黄嘌呤氧化酶的超氧阴离子生成系统、dpph清除实验和酵母细胞作为氧化应激耐受细胞模型来评估其抗氧化活性。结果:完整蛋白(EYsP)具有抗氧化活性,而胃蛋白酶水解产物(P-EYsP)具有较强的超氧化物清除活性,而P-EYsP- t、P-EYsP-α和P-EYsP- t /α缺乏超氧化物清除活性。P-EYsP及其后续酶(P-EYsP- t、P-EYsP-α和P-EYsP- t /α)均表现出显著的DPPH还原活性,但P-EYsP- t、P-EYsP-α和P-EYsP- t /α还原DPPH的活性最强。MALDI-TOF-MS分析发现了5种主要的抗氧化肽,其中2种来自蛋黄糖蛋白40(859、883Da), 2种来自脂卵磷脂(901Da、945 Da), 1种来自活蛋白(1089 Da)。该多肽具有清除超氧阴离子和DPPH的活性,显著增强了酵母细胞对过氧化物诱导的氧化应激的耐受性。结论:研究结果表明,这些生物活性肽在预防和对抗氧化应激相关疾病方面具有潜在的营养价值。关键词:蛋黄蛋白;生物活性肽;抗氧化剂;superoxide-scavenging;DPPH-reduction;酵母宽容
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Identification of potent antioxidant bioactive peptides from the soluble proteins of chicken egg yolk
Background: Eggs are an excellent  nutrient-dense food containing proteins, fats, carbohydrates, minerals, and vitamins. While many proteins are present in egg yolk, there arefew studies on their health benefits. Objective: The aim of this study was to explore the antioxidant peptides derived from the soluble protein fractions of egg yolk. Lipids and lipoproteins were removed with activated carbon and centrifugation after water dilution at acidic pH. Materials and Methods: Compared with 3.6 grams of protein in egg white, egg yolk contains 2.7 grams of protein in  a single large egg. The egg yolk soluble protein (EYsP) was digested with pepsin for 2h at pH 3.0 (P-EYsP), followed by digestion with trypsin (P-EYsP-T), α-chymotrypsin (P-EYsP-α), or both enzymes (P-EYsP-T/α). The most active digest was fractionated using sephacryl S-100 gel filtration, followed by reversed phase-HPLC of the most active fraction. The antioxidant activities were evaluated using a superoxide anion-generating system of xanthine oxidase, DPPH-scavenging assay, and yeast cells as an oxidative-stress tolerance cellular model. Results: The intact proteins (EYsP) showed antioxidant activity, but pepsin hydrolysate (P-EYsP) exhibited greater superoxide-scavenging activities than EYsP, while P-EYsP-T, P-EYsP-α and P-EYsP-T/α lacked activities. The P-EYsP and its subsequent proteases (P-EYsP-T, P-EYsP-α and P-EYsP-T/α) exhibited significant DPPH reduction, but P-EYsP-T, P-EYsP-α and P-EYsP-T/α exhibited the strongest DPPH reduction activities. MALDI-TOF-MS analysis revealed five major antioxidant peptides, two derived from yolk glycoprotein 40 (859 Da, 883Da), two from lipovitelline (901Da, 945 Da), and one from livetin (1089 Da). The peptides exhibited potent superoxide anion as well as DPPH scavenging activities and markedly enhanced the tolerance of yeast cells against peroxide-induced oxidative stress.Conclusion: The results show that these bioactive peptides hold a fascinating opportunity for their potential as nutraceuticals in prevention and combating oxidative stress-associated diseases.Keywords: Egg yolk proteins; bioactive peptides; antioxidant; superoxide-scavenging; DPPH-reduction; yeast tolerance
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