天然产物研究与开发 ›› 2020, Vol. 32 ›› Issue (8): 1348-1356.doi: 10.16333/j.1001-6880.2020.8.010

• 研究简报 • 上一篇    下一篇

阿胶多肽的高分辨质谱鉴定及活性研究

熊雅茹1,2,3,4，傅红1,4，杨方2,3*   

  1. 1福州大学生物科学与工程学院，福州 350108；2福州海关技术中心；3福建省检验检疫技术研究重点实验室，福州 350001；4福州大学 福建省海洋酶工程重点实验室，福州 350108

  • 出版日期:2020-08-28 发布日期:2020-09-02
  • 基金资助:
    国家重点研发专项（2017YFF0211000）

Identification and activity study of donkey-hide gelatin polypeptide by high resolution mass spectrometry

XIONG Ya-ru1,2,3,4,FU Hong1,4,YANG Fang2,3 *   

  1. 1College of Biological Sciences and Engineering,Fuzhou University,Fuzhou 350108,China;2Technology Center of Fuzhou Customs;3Fujian Key Laboratory for Technology Research of Inspection and Quarantine,Fuzhou 350001,China;4Fujian Provincial Key Laboratory of Marine Enzyme Engineering,Fuzhou University,Fuzhou 350108,China

  • Online:2020-08-28 Published:2020-09-02

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摘要：

建立了液相色谱-高分辨质谱分析阿胶中功能肽的方法，并进行了体外活性验证研究。阿胶经炖化、胰蛋白酶酶解，C18 SPE柱净化，以UPLC-Q-Exactive四极杆-静电场轨道阱高分辨质谱联用仪检测，MaxQuant软件分析质谱结果，所得肽段在UniProt上进行BLAST序列对比鉴定。结果表明，3次重复分析得到的63条共有肽中，具有明确蛋白及种属来源的肽19条。根据多肽活性构效关系，选择9条可能具有活性的阿胶多肽进行抗氧化活性、抗凝血活性测定，结果显示9条多肽均具有不同程度的抗氧化活性，其中5条具有抗凝血活性。本文所建立的方法快速有效，为揭示阿胶多肽结构与活性的关系提供技术支持。

关键词: 阿胶多肽, 炖化, 抗氧化活性, 抗凝血活性, 高分辨质谱

Abstract:

In this paper,a method for the identification of functional peptides in donkey-hide gelatin by liquid chromatography-high resolution mass spectrometry was established.In addition,the in vitro activity verification was carried out.After stewing,trypsin hydrolysis and purification with C18 SPE column,donkey-hide gelatin was detected by UPLC-Q-Exactive quadrupole-electrostatic field orbital trap high resolution mass spectrometer,and analyzed by MaxQuant software.The peptide segments were identified by BLAST sequence comparison on UniProt.The results showed that among 63 consensus peptides obtained from three repeated analyses,there were 19 peptides with clear protein and species origin.According to the structure-activity relationship of peptide activity,nine peptides that may be active were selected for antioxidant activity and anticoagulant activity determination.The results showed that nine peptides all had different degrees of antioxidant activity,of which five of them had anticoagulant activity.The method established in this paper is rapid and effective,and provides technical support for revealing the relationship between the structure and activity of donkey-hide gelatin polypeptide.

Key words: donkey-hide gelatin polypeptide, stewing, antioxidant activity, anticoagulant activity, high resolution mass spectrometry

中图分类号:  TS207.3

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