马面鱼皮ACE抑制肽的制备、分离纯化及稳定性
作者:
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(1.浙江万里学院生物与环境学院 浙江宁波 315100;2.上海海洋大学食品学院 上海 201306)

作者简介:

蔡金秀(1995—),女,硕士生

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基金项目:

浙江省重点研发项目(2019C02071);浙江省公益技术应用研究项目(GN22C206180)


Preparation, Isolation and Stability of Collagen ACE Inhibitory Peptides from the Skin of Navodon Septentrionalis
Author:
Affiliation:

(1.College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, Zhejiang;2.College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306)

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    摘要:

    为实现马面鱼皮的高值化利用,优化其胶原蛋白制备ACE抑制肽的酶解条件,通过超滤、凝胶色谱、高效液相及UPLC-MS等方法分离、纯化及分析ACE抑制肽。探讨pH值、温度及体外模拟胃肠消化对分离制备多肽的ACE抑制活性的影响。结果表明:利用复合酶解法可制备高活性ACE抑制肽,即在蛋白酶 N“天野”2G用量1 200 U/g,胰蛋白酶用量2 400 U/g,底物质量分数4%,温度40 ℃,pH 8的条件下酶解4 h,酶解产物ACE抑制率达74.3%。经超滤、凝胶分离制备3个高活性级分(a1,a2,a3),稳定性研究表明这3个级分在酸性和弱碱性条件下能保持较高活性,热稳定性好,经胃肠模拟消化后仍能保持较高的ACE抑制活性,其中级分a1的ACE抑制活性和稳定性最佳。HPLC及UPLC-MS分析表明,级分a1纯度较高,其氨基酸序列可能为Gly-Val-Ala或Ala-Val-Gly。本研究结果为马面鱼皮的精深加工及新型ACE抑制剂的开发提供一定的理论依据。

    Abstract:

    In order to realize the high value utilization of the skin of navodon septentrionalis, the optimal enzyme hydrolyzation technology for preparation of angiotensin converting enzyme (ACE) inhibitor peptide from collagen was studied. The ACE inhibitory peptides was separated, purified and analyzed by ultrafiltration, gel chromatography, high performance liquid chromatography (HPLC) and UPLC-MS. In addition, the effect of the pH, temperature and simulating gastrointestinal digestion on the stability of the ACE inhibitory peptides were assessed. The result revealed that the optimal parameters for producing protein hydrolysates with the highest ACE inhibitory activity were as follows: hydrolysis time 4 h at pH 8, Protease N “Amano” 2Genzyme dosage 1 200 U/g as well as trypsin enzyme dosage 2 400 U/g, hydrolysis temperature 40 ℃, and the enzyme-to-substrate ratio 4%. Under these conditions, the ACE inhibitory rate of enzymatic hydrolysates could reach 74.3%. After ultrafiltration and gel filtration, three high activity component (a1, a2, a3) were obtained, the stability study showed that the three high activity component(a1, a2, a3) maintained high activity under the thermal, acidic, mildly alkaline condition and demonstrated good stability in gastrointestinal digestion environment. Among them, a1 showed the best ACE inhibitory activity and stability. HPLC and UPLC-MS analysis showed that the purity of a1 was high, which amino acid sequence might be Gly-Val-Ala or Ala-Val-Gly. The above results provide a theoretical basis for the intensive processing of navodon septentrionalis skin and the development of new ACE inhibitors.

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蔡金秀,夏姗姗,马佳雯,王佳圆,曹少谦,戚向阳.马面鱼皮ACE抑制肽的制备、分离纯化及稳定性[J].中国食品学报,2022,22(3):225-234

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  • 收稿日期:2021-03-17
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  • 在线发布日期: 2022-04-13
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