Please wait a minute...
 
 
食品与发酵工业  2021, Vol. 47 Issue (5): 35-42    DOI: 10.13995/j.cnki.11-1802/ts.025510
  研究报告 本期目录 | 过刊浏览 | 高级检索 |
青鱼肉活性肽的制备及其抗肿瘤活性研究
张耀1, 张露1, 刘俊1, 涂宗财1,2*
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)
Preparation and antitumor activity of peptides from Mylopharyngodon piceus muscle
ZHANG Yao1, ZHANG Lu1, LIU Jun1, TU Zongcai1,2*
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)
下载:  HTML   PDF (1941KB) 
输出:  BibTeX | EndNote (RIS)      
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。
服务
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章
张耀
张露
刘俊
涂宗财
关键词:  青鱼肉  活性肽  二步酶解  抗氧化活性  抗肿瘤活性    
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
收稿日期:  2020-08-27      修回日期:  2020-09-23                发布日期:  2021-03-31      期的出版日期:  2021-03-15
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)
作者简介:  硕士研究生(涂宗财教授为通讯作者, E-mail:Tuzc_mail@aliyun.com)
引用本文:    
张耀,张露,刘俊,等. 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.
ZHANG Yao,ZHANG Lu,LIU Jun,et al. Preparation and antitumor activity of peptides from Mylopharyngodon piceus muscle[J]. Food and Fermentation Industries, 2021, 47(5): 35-42.
链接本文:  
http://sf1970.cnif.cn/CN/10.13995/j.cnki.11-1802/ts.025510  或          http://sf1970.cnif.cn/CN/Y2021/V47/I5/35
[1] JANSER S D C R, SATO H H.Biologically active peptides:Processes for their generation, purification and identification and applications as natural additives in the food and pharmaceutical industries[J].Food Research International, 2015, 74:185-198.
[2] MOHANTY D P, MOHAPATRA S, MISRA S, et al.Milk derived bioactive peptides and their impact on human health-A review[J].Saudi Journal of Biological Sciences, 2016, 23(5):577-583.
[3] KATHLEEN M, HEHIR, et al.A potential antitumor peptide therapeutic derived from antineoplastic urinary protein[J].Peptides, 2004, 25(4):543-549.
[4] SULERIA H A R, OSBORNE S, MASCI P, et al.Marine-based nutraceuticals:An innovative trend in the food and supplement industries[J].Marine Drugs, 2015, 13(10):6 336-6 351.
[5] 黄春红, 曾伯平, 董建波.青鱼、草鱼、鲢鱼和鳙鱼鱼头营养成分比较[J].湖南文理学院学报(自然科学版), 2008, 20(3):46-48.
HUANG C H, ZENG B P, DONG J B.Comparison of nutrients in the head of black carp, grass carp, silver carp and bighead carp[J].Journal of Hunan University of Arts and Science(Science and Technology), 2008, 20(3):46-48.
[6] 阎欲晓, 粟桂娇, 李小梅, 等.文蛤蛋白抗氧化活性肽的研究[J].食品工业科技, 2007, 28(12):121-123.
YAN Y X, LI G J, LI X M, et al.Study on the antioxidative active peptide of Meretrix meretrix[J].Science and Technology of Food Industry, 2007, 28(12):121-123.
[7] 杨林, 范美华, 刘雪珠, 等.两种合成抗菌肽的结构及抗菌作用机理[J].生物工程学报, 2011, 27(11):1 564-1 573.
YANG L, FAN M H, LIU X Z, et al.Solution structure and antibacterial mechanism of two synthetic antimicrobial peptides[J].Chinese Journal of Biotechnology, 2011, 27(11):1 564-1 573.
[8] 夏龙, 欧阳晓晖, 苏秀兰.抗菌肽及抗癌生物活性肽抗肿瘤机制研究进展[J].世界最新医学信息文摘, 2015(8):58-62.
XIA L, OUYANG X H, SU X L.Advance in the anti-cancer mechanism of antimicrobial peptides and anti-cancer biological active peptide[J].World Latest Medicine Information, 2015(8):58-62.
[9] RAJAPAKSE N, MENDIS E, BYUN H, et al.Purification and in vitro antioxidative effects of giant squid muscle peptides on free radical-mediated oxidative systems[J].The Journal of Nutritional Biochemistry, 2005, 16(9):562-569.
[10] JUNG W K, RAJAPAKSE N, KIM S.Antioxidative activity of a low molecular weight peptide derived from the sauce of fermented blue mussel, Mytilus edulis[J].European Food Research & Technology, 2005, 220(5-6):535-539.
[11] CHALAMAIAH M, YU W L, WU J P.Immunomodulatory and anticancer protein hydrolysates (peptides) from food proteins:A review[J].Food Chemistry, 2018, 245:205-222.
[12] DO N, WEINDL G, GROHMANN L, et al.Cationic membrane-active peptides-anticancer and antifungal activity as well as penetration into human skin[J].Experimental Dermatology, 2014, 23(5):326-331.
[13] LIU M, MIN L, ZHU C, et al.Preparation, characterization and antioxidant activity of silk peptides grafted carboxymethyl chitosan[J].International Journal of Biological Macromolecules, 2017, 104:723-738.
[14] 毛楷林. 皱瘤海鞘抗氧化及抗肿瘤组分的研究[D].海口:海南大学, 2018.
MAO K L.Study of antioxidant and anti-tumor components of Styela plicata[D].Haikou:Hainan University, 2018.
[15] 李军.鲢鱼骨胶原多肽的制备及其抗氧化、钙螯合活性的研究[D].南昌:江西师范大学, 2020.
LI J.Study on the preparation and antioxidant, calcium chelating activities of collagen polypeptides from silver carp bone[D].Nanchang:Jiangxi Normal University, 2020.
[16] 李军, 罗娟, 涂宗财, 等.鲢鱼骨胶原多肽的制备及其抗氧化活性研究[J].食品与发酵工业, 2020, 46(2):222-230.
LI J, LUO J, TU Z C, et al.Study on the preparation and antioxidant activity of collagen polypeptide from silver carp bone[J].Food and Fermentation Industries, 2020, 46(2):222-230.
[17] 涂宗财, 唐平平, 郑婷婷, 等.响应面优化鱼鳔胶原肽制备工艺及其抗氧化活性研究[J].食品与发酵工业, 2017, 43(5):160-166.
TU Z C, TANG P P, ZHENG T T, et al.Optimization of swimming bladder collagen peptide preparation using response surface methodology and its antioxidant activity research[J].Food and Fermentation Industries, 2017, 43(5):160-166.
[18] 罗艳华, 王全杰, 陈沛海, 等.蛋白水解物水解度测定方法的研究[J].皮革与化工, 2017, 34(2):26-31.
LUO Y H, WANG Q J, CHEN P H, et al.Study on determination method for hydrolyzed degree of protein hydrolysate[J].Leather and Chemicals, 2017, 34(2):26-31.
[19] CAI L, WU X, ZHANG Y, et al.Purification and characterization of three antioxidant peptides from protein hydrolysate of grass carp (Ctenopharyngodon idella) skin[J].Journal of Functional Foods, 2015, 16:234-242.
[20] 刘媛.海湾扇贝酶解物的制备、鉴定及其体内外抗肿瘤作用研究[D].保定:河北农业大学, 2015.
LIU Y.Preparation and Identification of the Argopecten irradias hydrolysate and its anti-tumor effects in vivo and in vitro[D].Baoding:Agricultural University of Hebei, 2015.
[21] 王文, 黄继红, 杨铭乾, 等.玉米胚芽蛋白水解物的制备及其抗氧化活性研究[J].食品工业, 2016, 37(6):47-52.
WANG W, HUANG J H, YANG M Q, et al.Study on preparation and antioxidant activity of hydrolysate from corn germ protein[J].The Food Industry, 2016, 37(6):47-52.
[22] 白仁奥.酶法制备文蛤活性多肽的研究[D].厦门:集美大学, 2013.
BAI R A.Study on the preparation of active peptides from Meretrix meretrix Linnaeu protein by enzymolysis[D].Xiamen:Jimei University, 2013.
[23] 肖群飞, 王天泽, 王羽桐, 等.鸡肉酶解物中不同氨基前体对肉香味形成的贡献[J].中国食品学报, 2020, 20(3):221-232.
XIAO Q F, WANG T Z, WANG Y T, et al.Roles of different amino-containing precursors in enzymatically hydrolyzed chicken in meat flavor formation[J].Journal of Chinese Institute of Food Science and Technology, 2020, 20(3):221-232.
[24] SONG R, ZHANG K Q, WEI R B.In vitro antioxidative activities of squid (Ommastrephes bartrami) viscera autolysates and identification of active peptides[J].Process Biochemistry, 2016, 51(10):1 674-1 682.
[25] 邹礼根, 郭水荣, 翁丽萍, 等.两种不同养殖模式对青鱼肌肉营养品质的影响[J].宁波大学学报(理工版), 2018, 31(4):25-30.
ZOU L G, GUO S R, WENG L P, et al.Effects of two different culture modes on muscle nutrients of black carp[J].Journal of Ningbo University (NSEE), 2018, 31(4):25-30.
[26] CHI C F, CAO Z H, WANG B, et al.Antioxidant and functional properties of collagen hydrolysates from spanish mackerel skin as influenced by average molecular weight[J].Molecules, 2014, 19(8):11 211-11 230.
[27] 胡小军, 江敏, 王标诗, 等.鱿鱼蛋白抗氧化肽的稳定性及抗疲劳和抗癌活性[J].食品工业科技, 2017, 38(16):60-64.
HU X J, JIANG M, WANG B S, et al.The stability of antioxidant peptides from squid and their anti-fatigue and anti-cancer activities[J].Science and Technology of Food Industry, 2017, 38(16):60-64.
[1] 冯艳钰, 臧延青. 三种小麦麸皮总黄酮的体外抗氧化活性[J]. 食品与发酵工业, 2021, 47(9): 16-24.
[2] 牛娜娜, 沙如意, 杨陈铭, 王珍珍, 茹语婷, 戴静, 韩洪庚, 张黎明, 毛建卫. 预处理工艺对黑蒜功能性成分、抗氧化活性影响及相关性研究[J]. 食品与发酵工业, 2021, 47(8): 67-75.
[3] 王子涵, 向敏, 徐茂, 蒋和体. 响应面优化黑果腺肋花楸汁澄清工艺及其抗氧化活性评价[J]. 食品与发酵工业, 2021, 47(8): 189-196.
[4] 顾欣, 高涛, 刘梦雅, 丛之慧, 张诚雅, 肖乐艳, 李迪, 胡景涛. 梁平柚柚皮多糖的提取、结构解析及抗氧化能力研究[J]. 食品与发酵工业, 2021, 47(7): 137-145.
[5] 匡文玲, 李佳, 韩林, 蒋永波, 邱玲岚, 汪开拓, 王敏. 柠檬果汁主要水溶性成分分析及对高脂诱导L-02肝细胞氧化损伤影响的研究[J]. 食品与发酵工业, 2021, 47(5): 43-47.
[6] 邓永平, 车鑫, 艾瑞波, 刘晓兰, 辛嘉英, 王晓杰. 好食脉孢霉发酵产类胡萝卜素的鉴定、抗氧化性及稳定性研究[J]. 食品与发酵工业, 2021, 47(4): 15-20.
[7] 孙洁, 李燕, 施文正, 汪之和. 虾类生物活性肽的研究进展[J]. 食品与发酵工业, 2021, 47(4): 261-268.
[8] 徐珍珍, 于秋生, 陈天祥, 平向莉, 曾朱强, 程丹. 大米蛋白肽的制备与ACE抑制活性分析[J]. 食品与发酵工业, 2021, 47(3): 53-58.
[9] 陆娟, 谢东雪, 贺柳洋, 王月, 郑志艳. 洋甘菊多糖的分离纯化、性质结构及抗氧化活性分析[J]. 食品与发酵工业, 2021, 47(3): 72-78.
[10] 谢三都, 陈惠卿, 庄培荣, 洪家榕, 游利杰. 冲泡型灵芝白茶的制备及其茶汤的抗氧化活性[J]. 食品与发酵工业, 2021, 47(3): 135-142.
[11] 彭松林, 潘成磊, 康梦瑶, 李懿璇, 赵紫悦, 郑仁兵, 尚永彪. 卤烤鸭中类黑精的提取及其抗氧化活性与化学稳定性研究[J]. 食品与发酵工业, 2021, 47(2): 22-29.
[12] 江飞凤, 谭晓辉, 胡鹏刚, 潘雪梅, 闫锦. 超声-微波协同提取柚子皮多糖工艺优化及单糖组成、结构和抗氧化活性分析[J]. 食品与发酵工业, 2021, 47(2): 196-204.
[13] 董艺凝, 李煜, 黄开军, 贾胜, 宋之文. 富含黄酮低度滁菊浸泡酒研制及品质分析[J]. 食品与发酵工业, 2021, 47(2): 220-225.
[14] 成柯, 闫俊, 严晓雪, 袁方. 湖北地区不同品种蓝莓果汁加工品质特征及抗氧化活性评价[J]. 食品与发酵工业, 2020, 46(9): 146-151.
[15] 朱秀灵, 叶精勤, 盛伊健, 孔雯瑾, 陈廷然, 傅锡鹏, 戴清源. 体外模拟消化对苹果多酚及其抗氧化活性的影响[J]. 食品与发酵工业, 2020, 46(8): 63-71.
No Suggested Reading articles found!
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
版权所有 © 《食品与发酵工业》编辑部
地址:北京朝阳区酒仙桥中路24号院6号楼111室
本系统由北京玛格泰克科技发展有限公司设计开发  技术支持:support@magtech.com.cn