摘要
[目的]研究蜂蜜中蛋白质提取的有效方法,优化蜂蜜蛋白电泳条件。[方法]该试验使用丙酮、钨酸钠、硫酸铵、尿素-硫脲4种方法提取蜂蜜中的蛋白并进行比较,然后进行双向电泳分析,对影响双向蛋白电泳的IPG胶条的选择、蛋白质上样量、等电聚焦时间等主要因素进行优化。[结果]试验表明,采用硫脲法提取蜂蜜蛋白获得的蛋白含量最高,硫脲法和丙酮沉淀法获得的蛋白纯度最好,而硫脲法因干扰物质较少、易于裂解适于双向电泳技术,采用硫脲法提取的蜂蜜蛋白选择IPG胶条p H 5~8,蛋白上样量为150μg,等电聚焦时间达到32 000 V·h的情况下能充分地聚焦,获得的双向电泳图谱最佳,优化后的洋槐蜜双向电泳图谱上可检测到326个蛋白点,重复性和分辨率均较好。[结论]蜂蜜类含糖样品中使用硫脲法提取蛋白能减少糖类物质干扰。
[ Objective ] To research a effective method for protein extraction from honey, and to optimize the protein electrophoresis conditions of honey. [ Method] Protein was extracted from honey by acetone, sodium tungstate, ammonium sulfate and urea-thiourea. Their results were compared and two-dimensional eleetrophoresis was analyzed. [ Result ] Protein content was the maximum by thiourea method; protein purity was the highest in thiourea method and acetone precipitation method. Since thiourea method had few interfering substances, and was suitable for two-dimensional electrophoresis technology, we selected thiourca method to extract protein. The IPG adhesive tape was pH 5 - 8 ; loading quantity of protein was 150 I^g; isoelectric focusing time was 32 000V h. Under these conditions, the optimal two-dimensional electrophoreto- gram was obtained. A total of 326 protein spots were detected with relatively good repeatabihty and resolution. [ Conclusion ] Extracting protein from honey by thiourea method reduces the interference of glucose.
出处
《安徽农业科学》
CAS
2016年第3期85-87,95,共4页
Journal of Anhui Agricultural Sciences
基金
国家质检总局科技计划项目(2014IK087)
关键词
蜂蜜蛋白
双向电泳
蛋白提取
Honey protein
Two-dimensional electrophoresis
Protein extraction
