摘要
以竹荪子实体为原料,以异丙醇溶剂脱脂,采用酶解法提取粗蛋白,并进行了时间、pH、温度以及加酶量实验来确定综合最佳提取条件.通过聚乙二醇浓度、硫酸铵浓度、氯化钠浓度以及pH的单因素实验确定了萃取率最佳的双水相体系并将竹荪多糖与蛋白分离开,经膜分离透析方法后使用离子交换层析的方法进一步去除多余的盐,最终获得高纯度竹荪蛋白(纯度达99%以上).采用SDS-PAGE法测定了蛋白质的相对分子量,并对蛋白结构进行了质谱分析,结果表明:竹荪蛋白含有60种蛋白质及77个肽段.
Taking the fruiting body of dictyophora as the raw material,the oil was removed by isopropanol solvent degreasing treatment,the crude protein was extracted by enzymatic hydrolysis,and the parallel experiments with different time,pH,temperature&enzyme amount were carried out to determine the method with the best comprehensive conditions.Then the aqueous two-phase extraction was carried out.The aqueous two-phase system with the best extraction rate was determined by the single factor parallel experiment of sodium chloride concentration and pH,and the polysaccharide of dictyophora was separated from the protein.After membrane separation and dialysis,the excess salt was further removed by ion exchange chromatography,and finally the high-purity dictyophora protein(purity more than 99%)was obtained.After that,the relative molecular weight of the protein was determined by SDS-PAGE,and the protein structure was analyzed by mass spectrometry.The results showed that dictyophora protein contained 60 proteins and 77 peptides.
作者
李享
王心茹
张明浩
周莉
LI Xiang;WANG Xinru;ZHANG Minghao;ZHOU Li(School of Pharmaceutical Sciences,South-Central Minzu University 430070,Wuhan China)
出处
《中南民族大学学报（自然科学版）》
CAS
北大核心
2023年第2期187-195,共9页
Journal of South-Central University for Nationalities：Natural Science Edition
基金
国家自然科学基金资助项目(30471225)。
关键词
竹荪
蛋白质
分离纯化
质谱
Dictyophora
protein
purification
mass spectrometry
