王斌,杨绣荣,邬华威,何宇

• 1:浙江海洋大学食品与医药学院
• 2:宁波今日食品有限公司

摘要(Abstract)：

【目的】建立并优化金枪鱼精巢中鱼精蛋白的制备方法,并研究其肝素拮抗效应。【方法】通过单因素和响应面试验,优化金枪鱼鱼精蛋白的提取工艺,采用羧甲基-琼脂糖凝胶CL-6B(CM Sephaorse CL-6B)离子交换层析对金枪鱼鱼精蛋白粗品进行纯化,并通过坂口反应、Tricine-十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)电泳、氨基酸分析和肝素结合力-滴定法测定样品纯度和效价。【结果与结论】确定金枪鱼鱼精蛋白的提取最佳工艺条件为提取温度36℃、硫酸浓度0.4 mol/L、液料比为6∶1、提取时间65 min,在此条件下得率为12.51%±1.2%。采用CM Sephaorse CL-6B离子交换层析对金枪鱼鱼精蛋白粗品进行纯化,获得Ⅰ、Ⅱ、Ⅲ3个组分。坂口反应显示组分Ⅲ呈现阳性反应,Tricine-SDS-PAGE电泳表明其分子质量为6.5～21.1ku,氨基酸质量分数为76.90%±0.37%,碱性氨基酸质量分数为39.00%±0.27%,精氨酸质量分数为25.86%±0.32%,肝素结合力-滴定法测定结果显示组分Ⅲ的效价约为标品鱼精蛋白的2/3。以上结果证明组分Ⅲ为目标蛋白,即金枪鱼鱼精蛋白。

关键词(KeyWords)： 金枪鱼;精巢;鱼精蛋白;活性

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81673349);; 舟山市科技专项(2019c21015)

作者(Author): 王斌,杨绣荣,邬华威,何宇

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