参豉糖蛋白提取工艺优化及纤溶活性研究  

Study on Extraction Process Optimization and Fibrinolytic Activity of Glycoprotein from Ginseng-douchi

作　　者：宋吉鹏 孙银玲 丁纯洁 郑宏宇 赵娢 孙丹丹 陈丽艳 SONG Ji-peng;SUN Yin-ling;DING Chun-jie(Heilongjiang Academy of Traditional Chinese Medicine,Harbin,Heilongjiang 150036)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036

出　　处：《安徽农业科学》2023年第12期147-153,共7页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(U20A20400);现代农业产业技术体系建设专项(CARS-21);黑龙江省财政专项(CZKYF-2021A002);黑龙江省卫生健康委课题(20211313050162)。

摘　　要：[目的]对参豉中糖蛋白的提取工艺进行考察,并对其纤溶活性进行评价。[方法]以糖蛋白提取率为指标,通过单因素法考察缓冲液、提取温度、料液比、提取时间和提取次数对其的影响,并运用响应面法分析优化参豉糖蛋白的提取工艺,采用蒽酮-硫酸法测定糖蛋白中多糖含量,SDS-PAGE和Tricine-SDS-PAGE凝胶电泳确定糖蛋白的分子量,利用琼脂糖纤维蛋白平板法测定纤溶活性。[结果]缓冲液、提取时间和料液比对糖蛋白提取率影响较大;响应面法优化获得参豉糖蛋白的最佳提取工艺条件为Tris-Base-NaCl缓冲溶液(pH=8.04)、料液比1∶10(g∶mL),于4℃提取6 h,参豉糖蛋白提取率为4.85%,其中多糖含量为12.34%;SDS-PAGE、Tricine-SDS-PAGE凝胶电泳结果显示,参豉糖蛋白分子量为4.1~27.0 kD;与豆豉糖蛋白相比,参豉糖蛋白纤溶活性提高了40.6%。[结论]经验证利用响应面法分析测得参豉糖蛋白与模型预测值接近,可用于参豉糖蛋白的提取,且参豉糖蛋白具有较高的纤溶活性,为其进一步开发利用提供依据。[Objective]The extraction process of glycoprotein in ginseng-douchi was investigated,and its fibrinolytic activity was evaluated.[Method]Taking the glycoprotein extraction rate as an index,the effects of buffers,extraction temperature,solid-liquid ratio,extraction time and extraction times were investigated by one-way method,and optimize the extraction process of ginseng-douchi glycoprotein by the response surface method.The polysaccharide content in glycoproteins was determined by anthraconone-sulfuric acid method,the molecular weight of glycoproteins was determined by SDS-PAGE and Tricine-SDS-PAGE gel electrophoresis,and its fibrinolytic activity was determined by agarose fibrin plate method.[Result]The buffers,extraction time and solid-liquid ratio had a greater impact on glycoprotein extraction rate.The optimal extraction process conditions for obtaining ginseng-douchi glycoprotein were optimized by the response surface method:Tris Base NaCl buffer solution(pH=8.04),solid-liquid ratio 1∶10(g∶mL),and extracted at 4℃for 6 hours,the extraction rate of ginseng-douchi glycoprotein was 4.85%,and the polysaccharide content in ginseng glycoprotein was 12.34%.SDS-PAGE and Tricine-SDS-PAGE gel electrophoresis results showed that the molecular weight of ginseng-douchi glycoprotein was 4.1-27.0 kD;compared with ginseng-douchi glycoprotein,the fibrinolytic activity of ginseng-douchi glycoprotein increased by 40.6%.[Conclusion]After verification,the ginseng-douchi glycoprotein measured by the response surface method is close to the predicted value of the model,which can be used for the extraction of ginseng-douchi glycoprotein,and ginseng-douchi glycoprotein has a high fibrinolytic activity,which provides a basis for its application.

关 键 词：参豉 糖蛋白 提取工艺 优化 响应面法 蛋白质分子量 纤溶活性 

分 类 号：R284[医药卫生—中药学]