食品科学 ›› 0, Vol. ›› Issue (): 167-172.

• 生物工程 • 上一篇    下一篇

酶解浒苔蛋白制备ACE抑制肽

盘赛昆1,2,周鸣谦1,2,姚东瑞1,2,孙 惠2   

  1. 1.江苏省海洋生物技术重点建设实验室 2.淮海工学院食品工程学院
  • 收稿日期:2011-09-17 修回日期:2012-04-04 出版日期:2012-05-15 发布日期:2012-05-07
  • 通讯作者: 盘赛昆 E-mail:pskgx@yahoo.com.cn
  • 基金资助:

    江苏海洋生物技术重点实验室开放基金

Preparation of ACE Inhibitory Peptides by Enzymatic Hydrolysis of Protein from Enteromorpha clathrata

Sai-Kun PAN,   

  • Received:2011-09-17 Revised:2012-04-04 Online:2012-05-15 Published:2012-05-07
  • Contact: Sai-Kun PAN E-mail:pskgx@yahoo.com.cn

摘要: 为了开发利用绿潮藻类条浒苔中含量丰富的蛋白质,采用可见分光光度法测定酶解物对ACE的抑制作用,考察碱性蛋白酶、Alcalase和胰蛋白酶对条浒苔蛋白的水解作用及其水解产物的ACE抑制活性,筛选出Alcalase作为酶解条浒苔蛋白制备具有降血压活性酶解物的适宜水解酶。在单因素试验的基础上,采用响应面分析法对该酶的酶解条件进行优化。结果表明,最佳酶解条件为:料液比1:50、加酶量2982U/g pro、温度47.9℃、pH7.53、酶解时间90min,在该条件下,酶解物的ACE抑制率IC50值为0.66mg/mL。不同截留分子质量的组分中,分子质量范围在1~5kD,平均肽链长度为16的组分ACE抑制活性最强,模拟体外消化实验结果表明,分子质量小于10kD的组分具有一定的对消化酶的抗性。

关键词: 浒苔, 酶解, ACE抑制肽, Alcalase

Abstract: In order to exploit abundant proteins in the green algae Enteromorpha clathrata, Enteromorpha clathrata was enzymatically hydrolyzed and the angiotensin-converting enzyme (ACE) inhibitory activity of its hydrolysate was determined spectrometrically. Alcalase was found to be more suitable for the preparation of ACE inhibitory peptides from Enteromorpha clathrata proteins than alkaline protease and trypsin. Based on one-factor-at-a-time experiments, response surface methodology was employed to optimize the hydrolysis conditions. The optimal hydrolysis process conditions were solid/liquid ratio of 1:50, enzyme dose of 2982 U/g protein, hydrolysis temperature of 47.9 ℃, hydrolysis pH of 7.53 and hydrolysis duration of 90 min. Under these conditions, a hydrolysate with an ACE inhibitory IC50 of 0.66 mg/mL was obtained. The hydrolysate consisted of fractions with a MWCO (molecular weight cut-off) of 1-5 kD and the fraction with an average chain length of 16 exhibited the strongest ACE inhibitory activity. The fractions with molecular weight of lower than 10 kD showed resistance to gastrointestinal proteases in vitro.

Key words: Enteromorpha clathrata, enzymatic hydrolysis, angiotensin Ⅰ-converting enzyme (ACE) inhibitory peptide, alcalase

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