鲨鱼皮明胶水解肽的制备、分离纯化与抗氧化活性研究

Study on Purification,Characterization,Antioxidant Activity of Shark Skin Gelatin Hydrolysate

目的:基于鲨鱼皮明胶水解肽的抗氧化活性,制备、分离纯化高活性的抗氧化多肽。方法:分别使用5种商业蛋白酶水解鲨鱼皮明胶,测定其水解产物的DPPH自由基清除能力、ABTS自由基清除活性以及对Fe2+诱导卵黄脂蛋白多不饱和脂肪酸过氧化反应的抑制作用;利用SP-Sephadex C-25阳离子交换色谱、Sephadex G-50凝胶过滤色谱和C18反向高效液相色谱(HPLC)对酸性蛋白酶的水解产物进行分离纯化。结果:酸性蛋白酶水解产物具有最佳的抗氧化活性;利用SP-Sephadex C-25、Sephadex G-50和C18-HPLC对酸性蛋白酶的水解产物进行分离纯化,所得洗脱组分E2具有最强的清除DPPH自由基活性。经过ESI质谱分析,其多肽组分的主要分子质量为1356u。结论:本研究制备并分离纯化得到高活性的抗氧化多肽,为将来的生产应用提供理论依据。

Objective：The hydrolysate of shark skin gelatin shows antioxidant activity, the study aims to prepare and purify the antioxidant peptide with significant activity. Methods： Five different kinds of commercial protease were used to hydrolyze shark skin gelatin, and the antioxidant activities such as the abilities to scanvage DPPH radicals and ABTS radicals, as well as theinhibition to peroxidation of yolk lipoprotein polyunsaturated fatty acid induced by Fe （Ⅱ） were measured, respectively; SP-Sephadex C-25 cation exchange chromatography, Sephadex G-50 gel filtration chromatogra- phy and C18-HPLC were used to separate the gelatin hydrolysate. Results：The acid protease hydrolysate showed the best antioxidant activity. Furthermore, an eluted fraction E2 with the strongest activities of scanvaging DPPH radicals was obtained after a series of chromatography separation. The result of ESI mass spectrometry analysis indicated that the main components of fraction E2 was a peptide with molecular weight of 1356 Da. Conclusion： The prepared antioxidant pep- tide might have potential application in industry.