猴头菌抗氧化肽的分离纯化

Isolation and Purification of Antioxidant Peptides from Hericium erinaceus

比较高压蒸煮法和闪式提取法提取猴头菌(Hericium erinaceus)蛋白的提取率,闪式提取法的蛋白提取率为18.56%,高于高压蒸煮法的12.48%。以蛋白提取率为指标,采用单因素实验和正交实验优化闪式提取法的工艺,优化的工艺为电压180 v、料液比1∶27.5(g∶mL)、提取次数4次。在此条件下,猴头菌蛋白提取率为20.34%。将优化条件下获得的粗提物水溶液进行膜分离和凝胶过滤层析,得到3个抗氧化肽组分LEP-1、LEP-2、LEP-3,其对DPPH自由基和ABTS自由基的清除率分别为76.5%~86.2%和94.2%~96.8%,3个组分间无显著性差异;LEP-1对Fe离子的还原能力优于LEP-2和LEP-3。经推测LEP-1的氨基酸序列为Gly-Ser-Gly-Gly-Ser-Gly-Gly-Ser-Ala,LEP-2的氨基酸序列为Gly-Gly-Val-Gly-Ala-Pro,LEP-3的氨基酸序列为Leu-Ala-Gly-His。

Two protein extraction methods,high-pressure cooking method and flash extraction method,were compared for extraction rate of H.erinaceus proteins.The flash extraction method showed better results and was then optimized through single factor experiments and orthogonal design.The optimized process was as follows:extraction voltage 180 v,solid-liquid ratio 1∶27.5(g∶mL),extraction repeated four times.Using the optimized process,the protein extraction rate was 20.34%,and the resultant protein extract was further isolated and purified by membrane separation and gel filtration chromatography to yield three peptide components,LEP-1,LEP-2,and LEP-3.The three peptide components showed similar capacity in scavenging DPPH and ABTS free radicals,and their DPPH and ABTS scavenging rates were 76.5%-86.2%and 94.2%-96.8%,respectively.The ferric ion reducing power of LEP-1 was greater than that of the other two.The amino acid sequences of the three peptides were predicted to be Gly-Ser-Gly-Gly-Gly-Gly-Ser-Gly-Gly-Ser-Ala,Gly-Gly-Val-Gly-Ala-Pro and Leu-Ala-Gly-His,respectively.