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Purification and characterization of antioxidant peptides from cooked eggs using a dynamic in vitro gastrointestinal model in vascular smooth muscle A7r5 cells
npj Science of Food ( IF 6.4 ) Pub Date : 2018-04-23 , DOI: 10.1038/s41538-018-0015-7
Jiapei Wang , Wang Liao , Chamila Nimalaratne , Subhadeep Chakrabarti , Jianping Wu

Antioxidant peptides derived from food sources are considered as safer alternatives to commercially available antioxidant drugs. As one of the most abundant protein sources, hen’s egg proteins were extensively used to produce antioxidant peptides by enzymatic hydrolysis. Our previous work indicated that gastrointestinal digestion of cooked eggs significantly increased the antioxidant activity due to hydrolysis of egg proteins. To characterize the responsible antioxidant peptides, cooked eggs were digested in a simulated in vitro model of human gastro-intestinal digestion. Prepared digests were fractionated with FPLC (Fast Protein Liquid Chromatography) and RP-HPLC (Reverse-Phase High-Performance Liquid Chromatography) and the antioxidant activity was determined in A7r5 cells (vascular smooth muscle cell line). Further identification of peptides from peptide fractions with the highest antioxidant activity was carried out using LC-MS/MS. Four peptides derived from ovalbumin, DSTRTQ (48–53), DKLPG (61–65), DVYSF (96–100), and ESKPV (205–209), were identified; of which DKLPG did not show antioxidant activity in cells. Enzyme cleave analysis suggested that these four peptides were likely released from ovalbumin only by pepsin non-specific cleaves. It is postulated that egg consumption may exert protection against oxidative stress on human health due to release of antioxidant peptides during digestion.



中文翻译:

使用动态体外胃肠道模型在血管平滑肌A7r5细胞中从煮熟的鸡蛋中纯化和鉴定抗氧化肽

食品来源的抗氧化剂肽被认为是市售抗氧化剂药物的更安全替代品。作为最丰富的蛋白质来源之一,鸡蛋蛋白被广泛用于通过酶促水解产生抗氧化剂肽。我们以前的工作表明,煮熟的鸡蛋经肠胃消化后,由于鸡蛋蛋白的水解而显着提高了其抗氧化活性。为了表征负责的抗氧化剂肽,将煮熟的鸡蛋在模拟的人体胃肠消化体外模型中进行消化。将制备的消化物用FPLC(快速蛋白液相色谱)和RP-HPLC(反相高效液相色谱)进行分级,并测定A7r5细胞(血管平滑肌细胞系)的抗氧化活性。使用LC-MS / MS从具有最高抗氧化活性的肽级分中进一步鉴定肽。从卵清蛋白,DSTRTQ(48-53),DKLPG(61-65),DVYSF(96-100)和ESKPV(205-209)中鉴定出四种肽;其中DKLPG在细胞中未显示抗氧化活性。酶裂解分析表明,这四种肽可能仅通过胃蛋白酶非特异性裂解而从卵白蛋白释放。据推测,由于消化过程中抗氧化剂肽的释放,食用鸡蛋可能会对人体健康产生氧化应激的保护作用。其中DKLPG在细胞中未显示抗氧化活性。酶裂解分析表明,这四种肽可能仅通过胃蛋白酶非特异性裂解而从卵白蛋白释放。据推测,由于消化过程中抗氧化剂肽的释放,食用鸡蛋可能会对人体健康产生氧化应激的保护作用。其中DKLPG在细胞中未显示抗氧化活性。酶裂解分析表明,这四种肽可能仅通过胃蛋白酶非特异性裂解而从卵白蛋白释放。据推测,由于消化过程中抗氧化剂肽的释放,食用鸡蛋可能会对人体健康产生氧化应激的保护作用。

更新日期:2018-04-23
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