食品科学 ›› 0, Vol. ›› Issue (): 0-0.

• 生物工程 •    下一篇

利用牡蛎制备DPP-IV抑制肽及其活性分析

曹敏杰   

  1. 集美大学
  • 收稿日期:2020-02-26 修回日期:2021-01-22 出版日期:2021-05-25 发布日期:2021-05-26
  • 通讯作者: 曹敏杰 E-mail:mjcao@jmu.edu.cn
  • 基金资助:
    鱼类脯氨酰内肽酶的结构、功能及其对胶原肽的降解机理;国家贝类产业技术体系

Preparation and Activity Analysis of DPP-IV Inhibitory Peptides from Pacific Oyster (Crassostrea gigas)

  • Received:2020-02-26 Revised:2021-01-22 Online:2021-05-25 Published:2021-05-26

摘要: 以牡蛎肉为原料制备二肽基肽酶-IV(DPP-IV)抑制肽。通过对比5种蛋白酶(木瓜蛋白酶、碱性蛋白酶、胰酶、中性蛋白酶和复合蛋白酶)制备的酶解液对DPP-IV的抑制活性,发现胰酶制备的酶解液对DPP-IV抑制效果最好。通过优化胰酶酶解条件,确定加酶量0.8%,pH 8.0,温度37?C,酶解时间90 min。酶解液经超滤、Sephadex G-15和反向高效液相色谱分离纯化,获得肽片段。通过质谱鉴定筛选得到两种肽段的序列为Glu-Ile-Thr-Ala-Leu-Ala-Pro-Ser-Thr-Met-Lys(EITALAPSTMK)和Ile-Leu-Ala-Pro-Pro-Glu-Arg(ILAPPER)。利用BIOPEP在线模拟分析了这2个肽的胃肠液消化特性。采用固相合成法合成肽段APSTM和ILAPPER,并应用于DPP-IV抑制活性研究。结果显示,APSTM和ILAPPER的IC50值分别为354.81和16.98 ?mol/L。本研究通过酶解牡蛎肉制备高抑制活性的DPP-IV抑制肽,为以牡蛎为原料的功能性食品开发利用提供了理论基础。

关键词: 牡蛎, 酶解, DPP-IV抑制肽, 分离纯化, 抑制活性

Abstract: Dipeptidyl peptidase-IV (DPP-IV) inhibitory peptides were prepared from Pacific oyster (Crassostrea gigas). By comparing the inhibitory activities of the hydrolysates prepared using 5 kinds of proteinases, including papain, alcalase, pancreatin, neutrase and protromex on DPP-IV, the pancreatin hydrolysate revealed the highest inhibitory activity. Optimal hydrolyzing condition was determined to be with 0.8% pancreatin, at pH 8.0, 37?C and hydrolysis for 90 min. The hydrolysate was further purified by ultrafiltration, Sephadex G-15 and reversed-phase high performance liquid chromatography (RP-HPLC). After purification, 2 peptides were obtained and identified as Glu-Ile-Thr-Ala-Leu-Ala-Pro-Ser-Thr-Met-Lys (EITALAPSTMK) and Ile-Leu-Ala-Pro-Pro-Glu-Arg (ILAPPER) by mass spectrometry. The digestibility characteristics of these two peptides in gastrointestinal fluid were analyzed using online simulation of BIOPEP. Two peptides APSTM and ILAPPER were synthesized by solid-phase synthesis. The results showed that IC50 values of APSTM and ILAPPER were 354.81 ?mol/L and 16.98 ?mol/L, respectively. Using oyster as raw material, peptides with effective DPP-IV inhibitory activity were prepared, which may provide a solid basis for comprehensive utilization of oyster in the future.

Key words: oyster, hydrolysis, DPP-IV inhibitory peptide, isolation and purification, inhibitory activity

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