食品与发酵工业 >

2021 , Vol. 47 >Issue 5: 35 - 42

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.025510

研究报告

青鱼肉活性肽的制备及其抗肿瘤活性研究

  • 张耀 ,
  • 张露 ,
  • 刘俊 ,
  • 涂宗财
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  • 1(江西师范大学 国家淡水鱼加工技术研发专业中心和江西省淡水鱼高值化利用工程技术研究中心, 江西 南昌, 330022)
    2(食品科学与技术国家重点实验室(南昌大学), 江西 南昌, 330047)
硕士研究生(涂宗财教授为通讯作者, E-mail:Tuzc_mail@aliyun.com)

收稿日期: 2020-08-27

  修回日期: 2020-09-23

  网络出版日期: 2021-03-31

基金资助

国家重点研发专项项目(2018YFD0901101);国家现代农业产业技术体系建设专项基金项目(CARS-45);江西省重点研发计划项目(20192ACB60005)

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Preparation and antitumor activity of peptides from Mylopharyngodon piceus muscle

  • ZHANG Yao ,
  • ZHANG Lu ,
  • LIU Jun ,
  • TU Zongcai
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  • 1(National R&D Center for Freshwater Fish Processing and Engineering Research Center for Freshwater Fish High-value Utilization of Jiangxi Province, Jiangxi Normal University, Nanchang 330022, China)
    2(State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China)

Received date: 2020-08-27

  Revised date: 2020-09-23

  Online published: 2021-03-31

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摘要

以青鱼肉为原料, 经二步酶解、凝胶色谱分离纯化制备抗氧化和抗肿瘤活性肽。首先以抗氧化活性和水解度为评价指标, 采用单因素和正交试验优化青鱼肉活性肽的制备工艺;然后采用凝胶色谱进行分离纯化, 得到不同分子质量多肽组分并分析其氨基酸组成、1, 1-二苯基-2-三硝基苯肼(1, 1-diphenyl-2-picrylhydrazyl, DPPH)自由基清除能力和抗肿瘤活性。结果表明, 二步酶解青鱼肉的最优酶解组合为碱性蛋白酶-复合蛋白酶, 最佳制备工艺为:pH 8.00、酶解温度50 ℃、料液比2∶10(g∶mL)、酶添加量6 000 U/g、酶解时间3 h, 在此条件的二步酶解物清除DPPH自由基的半抑制浓度(half inhibiting concentration, IC50)为9.52 mg/mL;经Sephadex G-15分离得到5个肽组分, 其分子质量范围为130~1 397 Da;抗氧化实验表明, 组分IV具有最强的DPPH自由基清除能力, 其IC50为3.17 mg/mL, 为二步水解物的3倍。细胞增殖抑制实验发现, 10 mg/mL组分IV对HepG2细胞抑制率为92.54%, 表明青鱼肉活性肽同时具有抗氧化和抗肿癌活性。

关键词： 青鱼肉; 活性肽; 二步酶解; 抗氧化活性; 抗肿瘤活性

本文引用格式

张耀 , 张露 , 刘俊 , 涂宗财 . 青鱼肉活性肽的制备及其抗肿瘤活性研究[J]. 食品与发酵工业, 2021 , 47(5) : 35 -42 . DOI: 10.13995/j.cnki.11-1802/ts.025510

Abstract

Antioxidant and antitumor peptides were prepared from Mylopharyngodon piceus muscle by two-step enzymolysis and gel chromatography separation. The preparation process was optimized by single factor and optimization experiment with antioxidant activity and degree of hydrolysis as indicators. Gel chromatography was used to separate and purify peptides with different molecular weights. The amino acid composition, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging capacity and antitumor activity of purified peptide fractions were also analyzed. The results showed that the optimum enzymatic hydrolysis conditions were: alkaline protease and composite protease, pH 8.00, 50 ℃, liquid-to-material ratio of 2∶10 (g∶mL), enzyme concentration of 6 000 U/g and hydrolysis time of 3 h. The half inhibitory concentration (IC50) value for DPPH free radical scavenging of sample prepared under optimal conditions was 9.52 mg/mL. Five peptides fractions (I-V) with molecular weights ranged from 130 to 1 397 Da were isolated by Sephadex G-15. Among which, fraction IV exhibited the highest scavenging ability on DPPH radicals, the IC50 value was 3.17 mg/mL, which was 3 times higher than that of the two-step hydrolysate. Cancer cell inhibition experiment showed that the inhibition rate of fraction IV (10 mg/mL) on HepG2 cells was 92.54%, indicating considerable antioxidant and antitumor activity of M. piceus muscle bioactive peptides. The results enrich the source of antitumor peptides and provides a technical reference for the high-value utilization of freshwater fish resources.

Key words： Mylopharyngodon piceus muscle; bioactive peptides; two-step enzymatic hydrolysis; antioxidant activity; antitumor activity

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