摘要
为研究霍山铁皮石斛糖蛋白的功效,以霍山铁皮石斛鲜茎为试验材料,比较盐溶液浸提和水提醇沉2种方法提取糖蛋白的效果。盐(NaCl)溶液浸提条件为:提取温度4℃、浓度0.3mol/L、料液比1∶16、提取时间24h,提取液经硫酸铵分级(15%、35%、60%)沉淀,去离子水透析脱盐后,再经聚乙二醇浓缩得糖蛋白提取液。水提醇沉提取条件:65℃水浴浸提,浸提3次,每次3h,对铁皮石斛进行浸提得提取液,加不同浓度(50%、70%、90%)乙醇进行沉淀得糖蛋白粗品,经去离子水复溶得糖蛋白溶液。2种方法所得的提取产物用SDS-PAGE电泳,再经蛋白染色和糖染色,确定并对比糖蛋白条带。结果表明:盐溶液浸提法获得的糖蛋白条带在浓度、数量和分子量范围上都显著高于水提醇沉法。试验结果为进一步开展霍山铁皮石斛糖蛋白的活性研究奠定了基础。
For the study of Mount Holyoke Dendrobium glycoprotein efficacy experiment on Mount Holyoke Dendro⁃bium as experimental materials,according to the glycoprotein extraction effect,comparative study of“salt leaching glycoprotein”and“water extraction and alcohol precipitation of glycoprotein”two methods.Salt(NaCl)solution ex⁃traction conditions:extraction temperature of 4 DEG C,the concentration of 0.3mol/L,solid-liquid ratio 1∶16,extrac⁃tion time 24h,extract by ammonium sulfate(15%,35%,60%)precipitation,deionized water purified by polyethyl⁃ene glycol,concentrating glycoprotein extract.Water extract alcohol extraction conditions:65 DEG C water bath ex⁃traction,extraction three times,each time 3h,extract of Dendrobium officinale extract liquid with different concentra⁃tions(50%,70%,90%)ethanol precipitated crude glycoprotein,dissolved by deionized water to glycoprotein solu⁃tion.The products were identified by 12%SDS-PAGE electrophoresis,and then stained and stained with glucose to determine and compare the glycoprotein bands.The results showed that the number and concentration of the glyco⁃protein bands obtained by salt solution extraction method were significantly higher than that of water extraction and alcohol precipitation method.The experimental results provide a basis for further research work activity of Dendrobi⁃um candidum glycoprotein in Mount Holyoke.
作者
祝启张
刘莉彬
崔绍进
刘庆慧
宁克壮
陈文正
张宁磊
邓辉
ZHU Qizhang;LIU Libin;CUI Shaojin;LIU Qinghui;NING Kezhuang;CHEN Wenzheng;ZHANG Ninglei;DENG Hui(College of Biological and Pharmaceutical Engineering,Luan 237012,China;West Anhui University Anhui Engi-neering Laboratory for Conservation and Sustainable Utilization of Traditional Chinese Medicine Resources,Luan 237012,China)
出处
《安徽农学通报》
2022年第2期24-26,共3页
Anhui Agricultural Science Bulletin
基金
2019年安徽省重点研究与开发计划项目(201904f06020007)
2018年度安徽省自然科学基金(1808085MH307)。
关键词
霍山铁皮石斛
糖蛋白
提取方法
Mount Holyoke Dendrobium
Glycoprotein
Extraction method
