Production and transepithelial transportation of casein-derived peptides and identification a novel antioxidant peptide LHSMK
Production and transepithelial transportation of casein-derived peptides and identification a novel antioxidant peptide LHSMK
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DOI：
10.1016/j.lwt.2021.112194
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发表时间：
2021-08-02
影响因子：
6
通讯作者：
Guo, Yuxing
中科院分区：
文献类型：
--
作者：
Liu, Mingzhen;Zhang, Tao;Guo, Yuxing
本研究利用胰蛋白酶水解酪蛋白衍生肽，然后通过Caco-2细胞单层转运，筛选肠道可吸收的抗氧化肽。通过响应面法确定了胰蛋白酶水解酪蛋白的最佳条件为：E/S = 5.45%，pH = 7.13，温度为51.99 ℃。将中空纤维超滤膜纯化的10 kDa以下组分应用于Sephadex G-10凝胶过滤层析，并将所得组分转运穿过Caco-2细胞单层。采用超高效液相色谱-串联质谱法在Caco-2细胞单层的基底外侧检测转运肽。发现LHSMK是一种未见报道的五肽。合成肽LHSMK具有清除自由基的能力，其抗氧化活性呈剂量依赖性增加。此外，LHSMK还可以调节氧化损伤细胞中抗氧化酶的活性。本研究为制备肠道可吸收肽提供了方法，为开发功能性食品市场提供了基础研究。
In this study, casein derived peptides were hydrolyzed with trypsin and then transported across Caco-2 cell monolayers to choose intestinal absorbable antioxidant peptides. E/S ratio at 5.45 %, pH at 7.13 and temperature at 51.99 degrees C were found to be the optimal conditions for enzymatic hydrolysis of casein by trypsin by Response Surface Method. The fraction below 10 kDa purified by the hollow fiber ultrafiltration membrane was applied to Sephadex G-10 gel filtration chromatography, and the obtained components were transported across the Caco-2 cell monolayer. The transported peptides were detected on the basolateral side of the Caco-2 cell monolayer using ultra-performance liquid chromatography-tandem mass spectrometry. It was found that LHSMK was an unreported pentapeptide. The synthetic peptide LHSMK had free radical scavenging ability, and the antioxidant activity exhibited a dose-dependent increase. Furthermore, LHSMK can also regulate antioxidant enzymes activity in oxidative-damage cells. This study provides a method for the preparation of intestinal absorbable peptides, and provides basic research for the development of functional food market.