摘要
以赤小豆凝集素为研究对象,采用响应面试验对其提取工艺进行优化。采用硫酸铵分级沉淀法、Q Sepharose XL阴离子交换色谱、Phenyl FF HP疏水层析色谱和Sephadex G-50凝胶过滤色谱进一步纯化,通过SDS-PAGE电泳对其蛋白质进行分析。结果表明,赤小豆凝集素的最优提取条件为:以PBS为浸提液,料液比1∶23(g/mL),pH 7.8,持续时间14 h,此条件下凝集活性为(37.18±0.18)HU;纯化后的凝集素凝集活性达127 HU,经鉴定目标凝集素具有两条亚基,分子量分别在55 kDa和27 kDa附近。
The Vigna umbellata lectin as the research object,based on the results of single factor experiments,the extraction process is optimized by response surface experiments.Further purification was made by using ammonium sulfate fractionation precipitation method,Q Sepharose XL anion exchange chromatography,phenyl FF HP hydrophobic chromatography and Sephadex G-50 gel filtration chromatography,and the protein was analyzed by SDS-PAGE electrophoresis.The results showed that the optimum extraction parameters were as follows:PBS is used as the extracting solution,the ratio of material to liquid was 1∶23(g/mL),pH value was 7.8,and the duration was 14 h and the agglutination activity of lectin was(37.18±0.18)HU.The agglutination activity of the purified lectin reached 127 HU.Identification of purified lectin has two subunits and the molecular weights were around 55 kDa and 27 kDa.
作者
肖俊琪
翟爱华
张东杰
XIAO Jun-qi;ZHAI Ai-hua;ZHANG Dong-jie(College of Food Science Heilongjiang Bayi Agricultural University,Daqing,Helongjiang 163319,China;Key Laboratory of Agro-Products Processing and Quality Safety of Heilongjiang Province,Daqing,Helongjiang 163319,China;National Coarse Cereals Engineering Research Center,Daqing,Helongjiang 163319,China)
出处
《食品与机械》
北大核心
2020年第7期161-166,200,共7页
Food and Machinery
基金
国家重点研发计划:(编号:2018YFE0206300)
黑龙江省优势特色学科资助项目:(编号:黑教联[2018]4号)。
关键词
赤小豆
凝集素
分离
纯化
电泳分析
Vigna umbellata
lectin
isolation
purification
electrophoresis analysis
