湖北农业科学 ›› 2024, Vol. 63 ›› Issue (11): 153-159.doi: 10.14088/j.cnki.issn0439-8114.2024.11.026

• 贮藏·加工 • 上一篇    下一篇

响应面优化超声波辅助酶解法制备金针菇ACE抑制肽

杨志雨, 王广慧, 孟涓涓, 郑晓宇, 徐丽晶, 韩昊霖   

  1. 绥化学院食品与制药工程学院,黑龙江 绥化 152061
  • 收稿日期:2023-05-08 出版日期:2024-11-25 发布日期:2024-12-03
  • 通讯作者: 王广慧(1973-),女,黑龙江绥化人,教授,硕士,主要从事食用菌中生物活性物质的分离与鉴定研究,(电话)18846509786(电子信箱)wanggh5487@sina.com。
  • 作者简介:杨志雨(2002-),女,黑龙江大庆人,在读本科生,专业方向为制药工程,(电话)18246833970(电子信箱)76575042@qq.com。
  • 基金资助:
    黑龙江省大学生创新创业训练计划项目(202210236006); 黑龙江省省属高等学校基本科研业务费基础研究项目(YWK10236200203)

Optimization of ultrasonic-assisted enzymatic hydrolysis for preparation of ACE inhibitory peptides from Flammulina velutipes by response surface methodology

YANG Zhi-yu, WANG Guang-hui, MENG Juan-juan, ZHENG Xiao-yu, XU Li-jing, HAN Hao-lin   

  1. College of Food and Pharmaceutical Engineering, Suihua University, Suihua 152061, Heilongjiang,China
  • Received:2023-05-08 Published:2024-11-25 Online:2024-12-03

摘要: 以金针菇(Flammulina velutipes)为原料,采用超声波辅助酶解法制备血管紧张素转化酶(ACE)抑制肽,为金针菇的深加工提供依据。筛选制备金针菇ACE抑制肽的酶,优化等电点沉淀法分离活性肽的最适pH。以ACE抑制率为响应值,开展液料比、超声时间、酶添加量等5个因素的单因素试验,设计四因素三水平的响应面试验。结果表明,制备金针菇ACE抑制肽的最佳蛋白酶为胰蛋白酶,沉淀活性肽的最佳pH为6.5。ACE抑制肽最佳制备条件为超声功率250 W,酶解温度37 ℃,酶解pH 8,液料比40∶1(mL/g),超声时间5 min,酶添加量为金针菇样品量的1.2%(g/g),酶解时间90 min。通过超滤分离法对ACE抑制肽进行了分级,并分析其活性。在此条件下,金针菇肽的ACE抑制率为(65.36±1.53)%;截留分子质量<3 kDa的肽ACE抑制率最高。

关键词: 金针菇(Flammulina velutipes), ACE抑制肽, 超声波辅助酶法, 响应面优化

Abstract: Angiotensin converting enzyme (ACE) inhibitory peptide was prepared from Flammulina velutipes by ultrasonic-assisted enzymolysis, providing the basis for further processing of Flammulina velutipes. The enzyme for preparing ACE inhibitory peptide of Flammulina velutipes was screened, and the optimal pH for separating active peptide by isoelectric point precipitation was optimized. With ACE inhibition rate as the response value, a single factor test was carried out with five factors, including liquid to material ratio, ultrasonic time, and enzyme dosage, and a response surface test with four factors and three levels was designed. The results showed that the best protease for preparing ACE inhibitory peptide of Flammulina velutipes was trypsin, and the best pH for precipitating active peptide was 6.5. The optimal preparation condition of ACE inhibitory peptide was as follows: ultrasonic power of 250 W, enzymolysis temperature of 37 ℃, enzymolysis pH of 8, liquid to material ratio of 40∶1 (mL/g), and ultrasonic time of 5 min. Enzyme addition was 1.2% (g/g) of Flammulina velutipes sample, and enzymolysis time was 90 min. ACE inhibitory peptides were classified and their activity was analyzed using ultrafiltration separation method. Under this condition, the ACE inhibitory rate of Flammulina velutipes peptide was (65.36 ± 1.53) %; peptides with a molecular weight of <3 kDa had the highest ACE inhibition rate.

Key words: Flammulina velutipes, ACE inhibitory peptide, ultrasonic-assisted enzymatic method, response surface optimization

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