食品科学 ›› 2012, Vol. 33 ›› Issue (24): 185-189.doi: 10.7506/spkx1002-6630-201224038

• 分析检测 • 上一篇    下一篇

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

邓丽莉1,潘晓倩1,生吉萍1,2,申琳1,*   

  1. 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
  • 收稿日期:2011-09-25 修回日期:2012-11-28 出版日期:2012-12-25 发布日期:2012-12-12
  • 通讯作者: 生吉萍 E-mail:pingshen@cau.edu.cn
  • 基金资助:

    苹果果实采后品质保持和调控机制;采后番茄果实抗病诱导过程中茉莉酸甲酯与一氧化氮协同作用机制

Optimization of Experimental Conditions for the Determination of Water Soluble Protein in Apple Pulp Using Coomassie Brilliant Blue Method

DENG Li-li1,PAN Xiao-qian1,SHENG Ji-ping1,2,SHEN Lin1,*   

  1. 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
  • Received:2011-09-25 Revised:2012-11-28 Online:2012-12-25 Published:2012-12-12

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。

关键词: 考马斯亮蓝法, 苹果, 微量可溶性蛋白, 吸光度

Abstract: The effects of extraction buffer type, pH and concentration, exogenous additive type and solid-to-solvent ratio on the extraction efficiency 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 influence extraction efficiency. 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 apples was 0.544% as determined using Coomassie brilliant blue method, a 7.45-fold increase as opposed to 0.073% with pure water extraction.

Key words: Coomassie brilliant blue method, apple, water soluble protein, absorbance

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