考马斯亮蓝法测定苹果组织微量可溶性蛋白含量的条件优化.pdfVIP

※分析检测 食品科学 2012, Vol.33, No.24 185 1 1 1,2 1,* 邓丽莉 ，潘晓倩 ，生吉萍 ，申 琳 (1. 中国农业大学食品科学与营养工程学院，北京 100083 ；2. 中国人民大学农业与农村发展学院，北京 100872) 摘 要：以红星苹果果肉为试材，探讨不同提取缓冲液(磷酸缓冲液、Tris-HCl缓冲液和水) 、缓冲液pH值、浓度、 外源添加物和料液比等因素对苹果果肉可溶性蛋白提取效率的影响。结果表明：外源添加PVP和EDTA 以及料液 比是苹果可溶性蛋白提取率的主要影响因素。实验最终确定的提取条件为0.1mol/L pH9.0 Tris-HCl提取缓冲液( 内 含1mmol/L EDTA和1% PVP)、料液比1:25，此条件下苹果果实可溶性蛋白的有效测定含量为0.544% ，为水提组 (0.073%)的7.45倍。 考马斯亮蓝法；苹果；微量可溶性蛋白；吸光度 Optimization of Experimental Conditions for the Determination of Water Soluble Protein in Apple Pulp Using Coomassie Brilliant Blue Method 1 1 1,2 1,* DENG Li-li ，PAN Xiao-qian ，SHENG Ji-ping ，SHEN Lin (1. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China； 2. School of Agricultural Economics and Rural Development, Renmin University of China, Beijing 100872, China) Abstract ：The effects of extraction buffer type, pH and concentration, exogenous additive type and solid-to-solvent ratio on the extraction effi ciency of soluble protein from Red Star apples were studied with the aim of developing a sample pretreatment method for the determination of water soluble protein in Red Star apples. Exogenous PVP and EDTA and solid-to-solvent ratio were found to be main factors that infl uence extraction effi ciency. The best results were obtained using 0.1 mol/L pH 9.0 Tris- HCl buffer containing 1 mmol/L EDTA and 1% PVP as an extraction solvent at a solid-to-solvent ratio of 1:25 (g/mL). Under these conditions, the content of water soluble protein in Red Star appl