摘要
本研究将菜籽蛋白通过碱性蛋白酶-风味蛋白酶分步酶解制备得到菜籽蛋白水解物(RPH),在经过超滤、葡聚糖凝胶柱(SephadexG-15)以及半制备液相(RP-HPLC)分离纯化得到各级分离组分。采用MTT比色法分析菜籽蛋白水解物各组分RPHs(MW<1ku)对人体肝癌细胞HepG2、乳腺癌细胞MBA-MD-231及人体结肠癌细胞Caco-2的体外抗增殖活性。当RPHs质量浓度为50~1000μg/mL时,RPHs对HepG2细胞、MBA-MD-231细胞及Caco-2细胞均有一定的抑制作用,且抑制效果与样品浓度之间呈现出剂量依赖性;其中,HepG2细胞对RPHs的作用最为敏感。经SephadexG-15凝胶柱分离后得到2个洗脱峰,收集并测定其活性,发现组分RPHs-F2具有更强的抗肿瘤细胞增殖活性;再进一步对其经半制备液相分离得到的组分进行体外抗增殖活性分析发现,除组分RPHs-F2-4外,其他3个分离组分都具有显著的抑制作用,而组分RPHs-F2-3对HepG2细胞具有最高的体外增殖抑制率,当作用质量浓度为1000μg/mL时,抑制率为(51.53±5.59)%。因此认为菜籽蛋白酶解物分离组分RPHs-F2-3具有较好的抗肿瘤活性,可以作为功能性成分用于抗肿瘤相关的功能食品及保健品的开发。
In this study,rapeseed protein was hydrolyzed by stepwise enzymatic hydrolysis (alkaline proteinase and flavourzyme).Each fraction of rapeseed protein hydrolysates was obtained by separating with ultrafiltration,gel column and semi-preparative HPLC.The proliferation of HepG2,MBA-MD-231,and Caco-2 cancer cells were also analyzed by MTT assay.When the RPHs (MW<1 ku) content was 50-1000μg/mL,RPHs inhibited HepG2 cells,mba-md-231 cells and caco-2 cells were in a dose-dependent manner.HepG2 cells were most sensitive to RPH.And the inhibition effect was dose-dependent with the concentration of the sample.Among them,HepG2 cancer cell showed the most sensitivity to the change of RPHs content.2 eluting peaks were obtained after the separation of Sephadex G-15 gel column.It was found that rphs-f2 had stronger anti-tumor cell proliferation activity upon collection and activity determination.Except for component RPHs-F2-4,the other three components showed significant inhibitory effects,while the component RPHs-F2-3 showed the highest in vitro inhibition of HepG2 cell proliferation.That is,when the action mass concentration was 1 000 g/mL,the inhibition rate was (51.53 5.59)%.Thus is can be seen that the separation of rapeseed protease hydrolysates RPHs-F2-3 has good anti-tumor activity,which can be used for the development of anti-tumor related functional food and health products as functional component.
作者
姚轶俊
袁强
鞠兴荣
王立峰
Yao Yijun;Yuan Qiang;Ju Xingrong;Wang Lifeng(Collaborative Innovation Center for Modern Grain Circulation and Safety/College of Food Science and Engineering,Nanjing University of Finance and Economics,Nanjing 210023;School of Food Science and Technology,Jiangnan University,Wuxi 214122)
出处
《中国粮油学报》
EI
CAS
CSCD
北大核心
2019年第7期47-53,共7页
Journal of the Chinese Cereals and Oils Association
基金
国家重点研发计划(2016YFD0401401)
南京市科技计划(201716034)
关键词
菜籽蛋白酶解物
分离纯化
抗肿瘤活性
rapeseed protein hydrolysate
separation and purification
antitumor activity
