食品科学 ›› 2020, Vol. 41 ›› Issue (24): 110-116.doi: 10.7506/spkx1002-6630-20191014-114

• 生物工程 • 上一篇    下一篇

条斑紫菜蛋白酶解液α-葡萄糖苷酶和DPP-IV抑制活性的表征及肽成分解析

黄钦钦,田亚平   

  1. (江南大学生物工程学院,江苏 无锡 214122)
  • 出版日期:2020-12-25 发布日期:2020-12-28
  • 基金资助:
    企业横向协作项目(180625)

α-Glucosidase and Dipeptidyl Peptidase IV Inhibitory Activity and Peptide Composition of Porphyra yezoensis Protein Hydrolysate

HUANG Qinqin, TIAN Yaping   

  1. (School of Biotechnology, Jiangnan University, Wuxi 214122, China)
  • Online:2020-12-25 Published:2020-12-28

摘要: 在特定可控酶解条件下制备获得条斑紫菜1 000 Da以下小分子多肽为主的蛋白酶解液,分析发现其具有对α-葡萄糖苷酶和二肽基肽酶IV(dipeptidyl peptidase IV,DPP-IV)的双酶抑制活性,且具有良好的pH值、温度稳定性和胃肠消化耐受性。将酶解液经超滤、纳滤、凝胶过滤层析分离纯化后,采用超高效液相色谱-电喷雾-四极杆飞行时间质谱对活性成分进行鉴定,解析出2 条多肽氨基酸序列为Ala-Pro、Pro-Gly-Gly-Val。按上述序列合成出这2 条多肽(纯度99.69%)表征其对2 种酶的抑制活性,结果表明:Pro-Gly-Gly-Val为双酶抑制肽,它对α-葡萄糖苷酶的IC50为18.60 mmol/L,对DPP-IV的IC50为17.80 mmol/L;而Ala-Pro只对DPP-IV有抑制作用,其IC50为4.65 mmol/L。

关键词: 条斑紫菜;酶解液特性表征;α-葡萄糖苷酶抑制活性;二肽基肽酶IV抑制活性;分离纯化;成分鉴定

Abstract: Porphyra yezoensis protein hydrolysate, prepared enzymatically under specific and controlled conditions, was found to be dominated by small peptides with molecular mass less than 1 000 Da. It had inhibitory activities against α-glucosidase and dipeptidyl peptidase IV (DPP-IV) and was able to tolerate pH, temperature and gastrointestinal digestion. The enzymatic hydrolysate was separated and purified consecutively by ultrafiltration, nanofiltration and gel filtration chromatography. Two bioactive peptides, Ala-Pro and Pro-Gly-Gly-Val, were identified by ultra-high performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (UPLC-ESI-Q-TOF-MS). Peptides corresponding to the sequences with a purity of 99.69% were separately synthesized and their inhibitory activities against the two enzymes were characterized. The results showed that Pro-Gly-Gly-Val was a double-enzyme inhibitory peptide with an IC50 of 18.60 mmol/L for α-glucosidase and of 17.80 mmol/L for DPP-IV. Ala-Pro had an inhibitory effect only against DPP-IV with an IC50 of 4.65 mmol/L.

Key words: Porphyra yezoensis; characterization of enzymatic hydrolysate; α-glucosidase inhibitory activity; dipeptidyl peptidase IV inhibitory activity; separation and purification; composition identification

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