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

作　　者：林塬 吴毓炜[2] 王焱 吉哲蓉 乐学义 LIN Yuan;WU Yuwei;WANG Yan;JI Zherong;LE Xueyi(Department of Applied Chemistry,South China Agricultural University,Guangzhou,Guangdong 510642,China;Products Quality Supervision and Testing Institute of Hainan Province/National Quality Supervision and Inspection Center for Tropical Agriculture Products,Haikou,Hainan 570203,China)

机构地区：[1]华南农业大学应用化学系,广东广州510642 [2]海南省产品质量监督检验所/国家热带农产品质量监督检验中心,海南海口570203

出　　处：《热带作物学报》2021年第4期1106-1112,共7页Chinese Journal of Tropical Crops

基　　金：海南省科学技术厅项目(No.KYYS-2014-69)。

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。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.

关 键 词：椰浆 蛋白分析 鉴定 RP-HPLC SDS-PAGE MALDI-TOF/TOF-MS 

分 类 号：S667.4[农业科学—果树学]