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目的:研究灰树花(Grifola frondosa)蛋白聚糖的提取与分离方法及氨基酸组成。方法:采用碱提法提取蛋白聚糖,QSepharose FF离子交换层析进行纯化,SDS-PAGE电泳分离蛋白聚糖及结合蛋白,高效液相色谱法鉴定蛋白聚糖的纯度及分子量,并测定18种氨基酸的含量。结果:分离出蛋白聚糖GFPⅠ、GFPⅡ和GFPⅢ,糖含量分别为:43.69%、40.88%、28.33%,蛋白含量分别为:40.42%、34.65%、51.19%。其中GFPⅢ为均一成分,纯度93.61%,重均相对分子量(Mr)150kDa,GFPⅢ结合蛋白分子量约为33.0kDa,为单一谱带。氨基酸组成分析结果显示:GFPⅢ的氨基酸总和达到66.60g/100g,其中8种必需氨基酸总量为26.14g/100g,4种呈味氨基酸总量为25.43g/100g,Asp、Glu、Leu、Ala和Lys含量较高。结论:分离出的蛋白聚糖为结构均一成分,纯度较高,可用于质量研究和生物学研究。
Objective: To study the extraction and isolation methods and amino acid composition of Grifola frondosa proteoglycan. Methods: Proteoglycan was extracted by alkali extraction, purified by QSepharose FF ion exchange chromatography, proteoglycan and binding protein were separated by SDS-PAGE electrophoresis, and the purity and molecular weight of proteoglycan were identified by HPLC. Amino acid content. Results: The proteoglycan GFPⅠ, GFPⅡ and GFPⅢ were isolated. The content of carbohydrate was 43.69%, 40.88%, 28.33%, respectively. The protein contents were 40.42%, 34.65%, 51.19% respectively. GFPⅢ was homogenous, with a purity of 93.61%. The weight average molecular weight (Mr) was 150 kDa. The molecular weight of GFP Ⅲ binding protein was about 33.0 kDa, which was a single band. The results of amino acid composition analysis showed that the total amino acids of GFP Ⅲ reached 66.60g / 100g, the total contents of 8 essential amino acids were 26.14g / 100g, the total contents of 4 kinds of amino acids were 25.43g / 100g, Asp, Glu, Leu, Ala and Lys content is higher. Conclusion: Proteoglycan isolated is a homogenous component with high purity, which can be used for quality research and biology research.