食品与发酵工业 >

2023 , Vol. 49 >Issue 7: 174 - 180

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.032219

研究报告

一种高纯度小分子解酒护肝肽的制备

  • 温庆仕 ,
  • 魏荷芬 ,
  • 周精卫 ,
  • 应汉杰 ,
  • 陈勇 ,
  • 刘庆国 ,
  • 袁童 ,
  • 刘淼 ,
  • 沈才洪 ,
  • 王松涛
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  • 1(江苏集萃工业生物技术研究所有限公司,江苏 南京, 211800)
    2(南京高新工大生物技术研究院有限公司,江苏 南京, 211800)
    3(南京工业大学 国家生化工程技术研究中心,江苏 南京, 210000)
    4(泸州老窖股份有限公司,四川 泸州,646000)
硕士,助理研究员(刘庆国助理研究员为通信作者,E-mail:liuqg1987@126.com)

收稿日期: 2022-05-05

  修回日期: 2022-06-23

  网络出版日期: 2023-04-28

基金资助

江苏省科协技术基金项目(BE2019001)

收起

Preparation of a high purity small molecule anti-alcoholic liver protecting peptide

  • WEN Qingshi ,
  • WEI Hefen ,
  • ZHOU Jingwei ,
  • YING Hanjie ,
  • CHEN Yong ,
  • LIU Qingguo ,
  • YUAN Tong ,
  • LIU Miao ,
  • SHEN Caihong ,
  • WANG Songtao
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  • 1(Jiangsu Jicui Industrial Biotechnology Research Institute Co.Ltd., Nanjing 211800, China)
    2(Nanjing Hi Tech University Biotechnology Research Institute Co.Ltd.,Nanjing 211800, China)
    3(National Biochemical Engineering Technology Research Center, Nanjing University of Technology, Nanjing 210000, China)
    4(Luzhou Laojiao Co.Ltd., Luzhou 646000, China)

Received date: 2022-05-05

  Revised date: 2022-06-23

  Online published: 2023-04-28

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摘要

目前海洋生物活性肽在功能食品中的应用已成为研究热点。针对中西药类解酒产品不足的问题,该研究制备了一种高纯度、高活性的小分子解酒护肝肽产品。采用多酶中度水解鱿鱼蛋白,以蛋白质回收率和乙醇脱氢酶(alcohol dehydrogenase,ADH)激活率为指标。确定了最佳酶解条件为胃蛋白酶∶碱性蛋白酶∶风味蛋白酶=1∶1∶1(质量比),酶添加量为0.2% (质量分数)、料液比为1∶10(g∶mL)、反应时间3 h。在最佳酶解条件下的酶解液,其蛋白回收率为80.34%,ADH激活率为83.37%。为进一步提纯小肽并提升其功能活性,对酶解液进行精制工艺研究,采用树脂吸附疏水性肽组分,用梯度乙醇洗脱,收集液浓缩、干燥,获得淡黄色产品肽粉末。其蛋白肽纯度为95.33%,其分子质量<3 000 Da的占99.96%,对ADH的酶活率为122.71%,对DPPH自由基的清除率可达92.01%。该研究制备的小肽产品在功能食品领域具有广阔的应用前景。

关键词： 多酶中度水解; 小分子肽; ADH激活率; DPPH自由基; 解酒护肝

本文引用格式

温庆仕 , 魏荷芬 , 周精卫 , 应汉杰 , 陈勇 , 刘庆国 , 袁童 , 刘淼 , 沈才洪 , 王松涛 . 一种高纯度小分子解酒护肝肽的制备[J]. 食品与发酵工业, 2023 , 49(7) : 174 -180 . DOI: 10.13995/j.cnki.11-1802/ts.032219

Abstract

At present, the research of marine bioactive peptides in functional foods has become popular. To solve the problem of the deficiency in traditional Chinese and Western anti-alcoholics, a small molecule anti-alcoholic liver protecting peptide with high purity and activity was prepared in this study. Squid protein was moderately hydrolyzed by multiple enzymes, and the protein recovery rate and alcohol dehydrogenase (ADH) activation rate were used as indicators. The optimum enzymatic hydrolysis conditions were∶pepsin∶alkaline protease∶flavor protease=1∶1∶1, enzyme addition 0.2% (mass fraction), solid-liquid ratio 1∶10 (g∶mL) and reaction time 3 h. Under the optimum conditions, the protein recovery rate was 80.34% and the ADH activation rate was 83.37%. In order to further purify the small peptide and improve its functional activity, the refining process of the enzymatic hydrolysate was studied. The hydrophobic peptide components were adsorbed by resin, eluted with gradient ethanol, and the collected solution was concentrated and dried to obtain a light yellow peptide powder. The purity of its protein peptide was 95.33%, peptide with the molecular weight under 3 000 Da was 99.96%, the ADH activation ratio was 122.71%, and the scavenging rate of DPPH free radical was 92.01%. The small peptide products prepared in this study have great application potential in the field of functional food.

Key words： moderate enzyme-complex hydrolysis; small molecule peptide; ADH activation rate; DPPH free radicals; protect liver therapy

参考文献

[1] YOKOYAMA H, BARAONA E, LIEBER C S.Upstream structure of human ADH7 gene and the organ distribution of its expression[J].Biochemical and Biophysical Research Communications, 1995, 216(1):216-222.
[2] 刘鹏. 低分子量促乙醇代谢玉米肽制备与蛋白成分对活性贡献关系研究[D].广州:华南理工大学,2015.
LIU P.Preparation of corn originated low-molecular-weight peptides with anti-alcoholism activity and the contribution of different protein components to activity[D].Guangzhou:South China University of Technology,2015.
[3] 赵谋明,马梅,苏国万,等.具有醒酒活性的玉米肽的制备、富集和鉴定[J].中国食品学报,2020,20(9):86-94.
ZHAO M M,MA M,SU G W, et al.Preparation, enrichment and identification of corn peptides with facilitating alcohol metabolism activity[J].Journal of Chinese Institute of Food Science and Technology, 2020,20(9):86-94.
[4] WU R B, WU C L, LIU D, et al.Overview of antioxidant peptides derived from marine resources:The sources, characteristic, purification, and evaluation methods[J].Applied Biochemistry and Biotechnology, 2015, 176(7):1 815-1 833.
[5] HOU Y Q, WU Z L, DAI Z L, et al.Protein hydrolysates in animal nutrition:Industrial production, bioactive peptides, and functional significance[J].Journal of Animal Science and Biotechnology, 2017, 8(1):24.
[6] HAYES M, MORA L.Alternative proteins as a source of bioactive peptides:The edible snail and generation of hydrolysates containing peptides with bioactive potential for use as functional foods[J].Foods (Basel, Switzerland), 2021, 10(2):276.
[7] WANG X Q, YU H H, XING R E, et al.Characterization, preparation, and purification of marine bioactive peptides[J].BioMed Research International, 2017, 2017:9746720.
[8] KIM S K, NGO D H, VO T S.Marine fish-derived bioactive peptides as potential antihypertensive agents[J].Advances in Food and Nutrition Research, 2012, 65:249-260.
[9] LIU D, HUANG J F, WU C L, et al.Purification, characterization, and application for preparation of antioxidant peptides of extracellular protease from Pseudoalteromonas sp.H2[J].Molecules (Basel, Switzerland), 2019, 24(18):3373.
[10] QIAN B J, ZHAO X, YANG Y, et al.Antioxidant and anti-inflammatory peptide fraction from oyster soft tissue by enzymatic hydrolysis[J].Food Science & Nutrition, 2020, 8(7):3 947-3 956.
[11] 叶昱辉. 近江牡蛎多肽的分离纯化及其抗氧化、抗光老化活性研究[D].广州:华南理工大学, 2018.
YE Y H.Purification of Ostrea rivularis peptides and their antioxidant, antiphotoaging activities[D].Guangzhou:South China University of Technology, 2018.
[12] DE J FIMBRES-ROMERO M, CABRERA-CHÁVEZ F, EZQUERRA-BRAUER J M, et al.Utilization of collagenolytic enzymes from sierra fish (Scomberomorus sierra) and jumbo squid (Dosidicus gigas) viscera to generate bioactive collagen hydrolysates from jumbo squid muscle[J].Journal of Food Science and Technology, 2021, 58(7):2 725-2 733.
[13] 江敏, 胡小军, 杨彩凤, 等.鱿鱼酶解工艺优化及酶解液抗氧化活性的研究[J].中国酿造, 2012, 31(6):143-146.
JIANG M, HU X J, YANG C F, et al.Optimization of enzymatic technology of squid and antioxidant activities of its hydrolyzate[J].China Brewing, 2012, 31(6):143-146.
[14] 江建梅. 鱿鱼酶解工艺及其酶解物的研究[D].天津:天津科技大学, 2010.
JIANG J M.Study on hydrolysis processing of squid and its hydrolysates[D].Tianjin:Tianjin University of Science & Technology, 2010.
[15] 刘铭, 刘玉环, 王允圃, 等.制备、纯化和鉴定生物活性肽的研究进展及应用[J].食品与发酵工业, 2016, 42(4):244-251.
LIU M, LIU Y H, WANG Y P, et al.Research progress on production, purification and identification of bioactive peptides and its challenges in application[J].Food and Fermentation Industries, 2016, 42(4):244-251.
[16] 林燕, 王鸿飞, 李和生, 等.鱿鱼皮酶解产物的功能特性[J].中国食品学报, 2011, 11(5):83-88.
LIN Y, WANG H F, LI H S, et al.Functional properties of enzymatic hydrolysates from squid skin[J].Journal of Chinese Institute of Food Science and Technology, 2011, 11(5):83-88.
[17] MANIATIS T.Molecular Cloning:A Laboratory Manual[M].Cold Spring Harbor:Cold Spring Harbor Laboratory Press, U.S., 1989.
[18] SHAVANDI A, HU Z H, TEH S, et al.Antioxidant and functional properties of protein hydrolysates obtained from squid pen chitosan extraction effluent[J].Food Chemistry, 2017, 227:194-201.
[19] SABLE R, PARAJULI P, JOIS S.Peptides, peptidomimetics, and polypeptides from marine sources:A wealth of natural sources for pharmaceutical applications[J].Marine Drugs, 2017, 15(4):124.
[20] 邹媛婷, 张健, 王共明, 等.生物活性肽在特殊医学用途配方食品中的应用研究进展[J].食品与发酵工业, 2022, 48(17):336-344.
ZOU Y T, ZHANG J, WANG G M, et al.Research progress on the application of bioactive peptides in Food for Special Medical Purpose[J].Food and Fermentation Industries, 2022, 48(17):336-344.
[21] 李富强, 张廷新, 朱丽萍, 等.食物蛋白源免疫调节肽研究进展[J].食品与发酵工业, 2022, 48(1):308-314.
LI F Q, ZHANG T X, ZHU L P, et al.Advance of immunomodulatory peptides from dietary-protein source[J].Food and Fermentation Industries, 2022, 48(1):308-314.
[22] HASEBA T, YAMAMOTO I, KAMII H, et al.Alcohol dehydrogenase (ADH) isozymes in the AdhN/AdhN strain of Peromyscus maniculatus (ADH-deermouse) and a possible role of class III ADH in alcohol metabolism[J].Biochemical Genetics.1995,33(9):349-363.
[23] 叶松, 任凌丽, 陈曦, 等.王氏保赤丸对急性酒精损伤小鼠醒酒护肝/胃作用及机制研究[J].中国中药杂志, 2021, 46(15):3 900-3 906.
YE S, REN L L, CHEN X, et al.Protective effect and mechanism of Wangshi Baochi Pills against acute alcoholic liver/stomach injury in mice[J].China Journal of Chinese Materia Medica, 2021, 46(15):3 900-3 906.
[24] 隋玉杰, 何慧, 石燕玲, 等.玉米肽的醒酒活性体外试验及其醒酒机理研究[J].中国粮油学报, 2008, 23(5):54-58.
SUI Y J, HE H, SHI Y L, et al.A study on mechanism and tests in vitro for corn peptides facilitating alcohol metabolism[J].Journal of the Chinese Cereals and Oils Association, 2008, 23(5):54-58.
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