双向电泳技术分离燕窝水溶性蛋白  

Applicat ion of Two-Dimensional Electrophoresis Technology in Separation of Water-Soluble Protein from Edible Bird's Nest

作　　者：郭丽丽[1,2] 吴亚君[1] 刘鸣畅[1] 王斌[1] 韩建勋[1,2] 葛毅强[2,3] 陈颖[1] 

机构地区：[1]中国检验检疫科学研究院,北京100123 [2]中国农业大学食品科学与营养工程学院,北京100083 [3]中国农村技术开发中心,北京100045

出　　处：《食品科学》2013年第24期97-101,共5页Food Science

基　　金："十二五"国家科技支撑计划项目(2013AA102202)

摘　　要：为研究燕窝功效成分以及建立燕窝真伪检测和质量分级体系,采用双向电泳技术分离了燕窝水溶性蛋白。以印度尼西亚苏门答腊岛毛燕为材料,从提取溶剂、提取条件、水化上样缓冲液和分离胶体积分数4个方面对燕窝蛋白双向电泳技术进行优化。结果表明:以超纯水作为提取溶剂,60℃静置提取6h,以8mol/L尿素、4g/100mL CHAPS、65mmol/L DTT、0.2%(体积分数)Bio-Lyte 4/6 Ampholyte、0.1%Bio-Lyte 5/8 Ampholyte、0.001g/100mL溴酚蓝组成的溶液作为水化上样缓冲液,采用8%的分离胶体积分数可获得高分离度、高重复性的燕窝蛋白双向电泳图谱。同时,采用4个不同的燕窝样品对所建立的方法进行验证,证实了方法的可靠性。In order to explore the functional components in edible bird＇s nest （EBN） and establish a system for the identification of EBN adulteration and quality classification, two-dimensional electrophoresis （2-DE） was adopted to separate water-soluble protein from EBN. EBN samples from the Indonesian island of Sumatra were analyzed by 2-DE and the assay was optimized with respect to extraction solvent, extraction conditions, rehydration buffer composition and resolving gel concentration. The results showed that 2-DE map of EBN protein with high resolution and good reproducibility could be obtained under the conditions： 6 h of extraction at 60 ℃ using ultrapure water as extraction solvent,, rehydration buffer composed of 8 mol/L urea, 4 g/100mL CHAPS, 65 mmol/L DTT, 0.2% Bio-Lyte 4/6 Ampholyte, 0.1% Bio-Lyte 5/8 Ampholyte and 0.001 g/100mL bromophenol blue, and separation gel concentration of 8%. Furthermore, four different EBN samples were separated successfully by the established 2-DE technology, showing the reliability of the developed method.

关 键 词：燕窝 蛋白质 提取 SDS-聚丙烯酰胺凝胶电泳 双向电泳 

分 类 号：TS201.2[轻工技术与工程—食品科学]