School of Medicine and Pharmacy and College of Food Science and Technology, Ocean University of China, Qingdao 266003, China.
School of Food Science and Technology, Qingdao Agricultural University, Qingdao 266109, China.
Mar Drugs. 2019 Jul 31;17(8):450. doi: 10.3390/md17080450.
The effect of collagen peptides (CPs) in intestinal mucosal protection has been approved in both cell and animal models. However, its structure-activity relationship and efficient peptide sequences are unclear, which hinders the in-depth study of its action mechanism and relative nutraceuticals and pharmaceuticals development. In this work, size exclusion chromatography, cation-exchange chromatography, and RP-HPLC were used to separate Alaska pollock skin-derived collagen hydrolysates based on their molecular weight, charge property, and hydrophobicity. The intestinal epithelial barrier function (IEBF) protective effect of separated peptide fractions were evaluated by tumor necrosis factor (TNF)-α-induced Caco-2 cell model. Results indicated that lower molecular weight (500-1000 Da) and higher hydrophilicity of CPs were related to better IEBF protective effect. Two high-efficiency IEBF protective peptide sequences, GPSGPQGSR and GPSGLLGPK with the corresponding molecular weights of 841.41 Da and 824.38 Da, were subsequently identified by UPLC-QToF-MS/MS. Their IEBF protective ability are comparable or even better than the currently used intestinal health supplements glutamine and arginine. The present findings suggested that the hydrophilic CPs, with molecular weight between 500 Da to 1000 Da, should be preferred in IEBF protective peptides preparation. GPSGPQGSR and GPSGLLGPK might have the potential of being IEBF protective ingredients used in intestinal health supplements and drugs.
胶原蛋白肽（CPs）在肠黏膜保护中的作用已在细胞和动物模型中得到证实。然而，其结构-活性关系和有效的肽序列尚不清楚，这阻碍了其作用机制的深入研究及其相关营养保健品和药物的开发。在这项工作中，使用排阻色谱、阳离子交换色谱和反相高效液相色谱（RP-HPLC）根据分子量、电荷性质和疏水性对阿拉斯加鳕鱼皮胶原水解物进行分离。采用肿瘤坏死因子（TNF）-α诱导的 Caco-2 细胞模型评价分离肽段的肠上皮屏障功能（IEBF）保护作用。结果表明，CPs 的低分子量（500-1000Da）和高亲水性与更好的 IEBF 保护作用相关。通过 UPLC-QToF-MS/MS 鉴定出两种高效 IEBF 保护肽序列 GPSGPQGSR 和 GPSGLLGPK，其分子量分别为 841.41Da 和 824.38Da。它们的 IEBF 保护能力与目前使用的肠道健康补充剂谷氨酰胺和精氨酸相当，甚至更好。本研究结果表明，在制备 IEBF 保护肽时，应优先选择分子量在 500Da 至 1000Da 之间的亲水性 CPs。GPSGPQGSR 和 GPSGLLGPK 可能具有作为肠道健康补充剂和药物中 IEBF 保护成分的潜力。
