三氯乙酸法测定烟叶中蛋白质含量  

Determination of protein content in tobacco leaves by trichloroacetic acid precipitation

作　　者：刘肖肖 樊新顺 施东青 田泱源 王艺霖 丁瑞红 王洪涛[1] 王文领 LIU Xiaoxiao;FAN Xinshun;SHI Dongqing;TIAN Yangyuan;WANG Yilin;DING Ruihong;WANG Hongtao;WANG Wenling(School of Life Science,Henan University,Kaifeng 475004,Henan,China;Henan Cigarette Industry（Xuchang）Tobacco Sheet Company Limited,Xuchang 461000,Henan,China;School of Life Science,Nanjing Agricultural University,Nanjing 210095,Jiangsu,China)

机构地区：[1]河南大学生命科学学院,河南开封475004 [2]河南卷烟工业(许昌)烟草薄片有限公司,河南许昌461000 [3]南京农业大学生命科学学院,江苏南京210095

出　　处：《化学研究》2018年第5期484-487,共4页Chemical Research

基　　金：中国烟草总公司标准预研项目(2013QB017)

摘　　要：烟叶中蛋白质的含量与烟草的品质息息相关.如何快速、准确地测定烟叶中蛋白质含量则显得尤为重要.本实验通过对烟叶蛋白质抽提液的最佳浓度、蛋白质沉淀剂的最佳浓度及加入量进行确定,探索出了一种适合于测定烟叶中蛋白质含量的方法:室温条件下,用浓度为8 mol·L-1的尿素溶液充分抽提烟叶中的蛋白质(抽提时间大于6 h),12 000 r/min离心20 min后仅留上清液,将所得上清液稀释4倍后加入3%三氯乙酸,静置10 min,充分沉淀上清液中的蛋白质,之后再次12 000 r/min离心20 min后弃上清,留沉淀,将沉淀加入丙酮清洗后烘干至恒重,达到恒重的沉淀质量即为该烟叶中蛋白质的质量.The content of protein in the cigarette affects its quality a lot. Therefore, how to determine the protein content in tobacco leaves quickly and accurately is particularly important. In this experiment, we explored a method suitable for determining protein content in tobacco leaves. The method was involved with different concentrations of urea in extraction buffer, different concentrations and amounts of trichloroacetic acid as a precipitant. The established system includes the following steps： extracting total protein from C3K tobacco leaf protein by 8 mol·L -1 urea solution dealing with for more than 6 h, centrifuging （ 12 000 r/min, 20 min）, diluting the supernatant to 4 times and then using 3% trichloroacetic acid to precipitate the protein, centrifuging （ 12 000 r/min, 20 min） again to receive the protein, washing the precipitate with acetone, lastly drying precipitate until it reaches constant weight. The quality of the precipitation is the quality of the protein in the tobacco leaf.

关 键 词：烤烟 蛋白测定 尿素 三氯乙酸 

分 类 号：Q816[生物学—生物工程]