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椰浆中椰子蛋白的提取、分离和鉴定
Extraction, Isolation and Identification of Coconut Protein from Coconut Milk
【摘要】 为了更好地了解市售椰浆中椰子蛋白质量状况,本研究以市售的10种椰浆为样品,采用反向高效液相色谱(reversed-phase high performance liquid chromatography,RP-HPLC)及十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(sodium laurylsulfonate-polyacrylamide gel electrophoresis,SDS-PAGE)对提取蛋白进行分析,采用基质辅助激光解吸电离飞行时间串联质谱(matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry,MALDI-TOF/TOF-MS)对蛋白进行鉴定。结果表明:椰浆RP-HPLC约在10.4 min和11.9 min分别出现1个色谱峰,大多数样品峰面积大小都与蛋白标签含量一致,SDS-PAGE条带数量和颜色的深浅与RP-HPLC得到的峰面积结果一致,个别样品标签含量高,SDS-PAGE能分离出的蛋白条带数量也较多,但RP-HPLC的椰子蛋白峰面积小,经MALDI-TOF/TOF-MS鉴定,部分条带没有鉴定出椰子有关的肽段或蛋白。研究结果为椰浆蛋白的食品安全与质量控制提供了方法参考和示范。
【Abstract】 In order to better understand the quality of coconut protein in commercially available coconut milk, 10 types of coconut milk were used as the samples bought from the market. Reversed-phase high performance liquid chromatography(RP-HPLC) and sodium laurylsulfonate-polyacrylamide gel electrophoresis(SDS-PAGE) were used to analyze the extracted proteins, identified through matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry(MALDI-TOF/TOF-MS). There were two peaks in RP-HPLC of coconut milk at 10.4 min and 11.9 min, the peak area of most samples were consistent with the protein label content. The number of SDS-PAGE bands and intensity were consistent with the peak area results obtained by RP-HPLC. Some samples had high label content, and SDS-PAGE could separate more protein bands, but the peak area of coconut protein from RP-HPLC was small. The protein bands were identified with MALDI-TOF/TOF-MS and no coconut-related peptides or proteins were identified. This method could be used for the isolation and identification of coconut protein in coconut milk.
【Key words】 coconut milk; protein analysis; identification; RP-HPLC; SDS-PAGE; MALDI-TOF/TOF-MS;
- 【文献出处】 热带作物学报 ,Chinese Journal of Tropical Crops , 编辑部邮箱 ,2021年04期
- 【分类号】TS201.21;O657
- 【被引频次】1
- 【下载频次】672