长萼堇菜叶蛋白水解物的制备及其活性分析  

Preparation and activity analysis of Viola inconspicua Blume leaf protein hydrolysates

作　　者：习雪玲 杨茗婷 覃婷 廖斌[3] 张军 XI Xueling;YANG Mingting;QIN Ting;LIAO Bin;ZHANG Jun(School of Biosciences and Biopharmaceutics,Guangdong Pharmaceutical University,Guangzhou,Guangdong 510006,China;Guangdong Province Key Laboratory for Biotechnology Drug Candidates,Guangzhou,Guangdong 510006,China;School of Life Science,Sun Yat-sen University,Guangzhou,Guangdong 510275,China)

机构地区：[1]广东药科大学生命科学与生物制药学院,广东广州510006 [2]广东省生物技术候选药物重点实验室,广东广州510006 [3]中山大学生命科学学院,广东广州510275

出　　处：《湖南农业大学学报（自然科学版）》2020年第6期716-722,共7页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(31570506);广东药科大学创新强校项目(2017KCXTD020)。

摘　　要：以长萼堇菜(Viola inconspicua Blume)为材料,优选碱提酸沉法参数,提取其叶蛋白;采用酶法制备叶蛋白水解物;利用超滤法分离叶蛋白水解物组分,分析叶蛋白水解物的抗氧化活性,并对叶蛋白水解物组分进行固相萃取脱盐、HPLC分析及其对肿瘤细胞的抑制活性和对线虫寿命影响的测试。结果表明:采用料液比、碱提pH和酸沉pH分别为1∶9、10.0和4.5的参数组合可稳定提取长萼堇菜叶蛋白;优选的3种蛋白酶(木瓜蛋白酶、碱性蛋白酶和风味蛋白酶)均能有效将水解物主要酶解为相对分子质量小于3.3×10^3的组分,并通过超滤可有效分离出相对分子质量分别为<1×10^3(PH1)、1×10^3~<3×10^3(PH2)和≥3×10^3(PH3)的3个组分;长萼堇菜蛋白水解物中PH1组分的抗氧化活性和抑肿瘤细胞活性优于其他组分;PH1和PH2延长线虫寿命的效果相近,都优于PH3。可见,长萼堇菜叶蛋白水解物组分PH1更具应用开发价值。Viola inconspicua Blume was purified using the optimized alkali extraction and acid precipitation method.Leaf protein hydrolysates were prepared by enzymatic method.The components of leaf protein hydrolysates were separated by ultrafiltration,and their antioxidant activities were analyzed.The isolated hydrolysate components were desalted by solid phase extraction,evaluated by HPLC analysis,and assayed the inhibitory activities to tumor cells along with the effects on lifespan of nematodes.V.inconspicua leaf proteins could be stably extracted with the optimized parameters(ratio of material to liquid 1∶9,pH value of alkali extraction 10.0,and pH value of acid precipitation 4.5).The three selected proteases(papain,basic protease and flavor enzyme)could make most of hydrolysates with relative molecular weight less than 3.3×10^3,and ultrafiltration could isolate the groups of hydrolysate components with molecular weight of<1×10^3(PH1),1×10^3-<3×10^3(PH2)and≥3×10^3(PH3),respectively.The antioxidant activity and anticancer activity of the component PH1 were better than those of other components.PH1 and PH2 had similar effects on lifespan of nematodes,but both were superior to PH3.Therefore,the component PH1 of Viola inconspicua leaf protein hydrolysates had more application and development value.

关 键 词：长萼堇菜 叶蛋白 蛋白水解物 秀丽隐杆线虫 生物学活性 

分 类 号：TQ936.1[化学工程] TQ464.7