食品科学 ›› 2005, Vol. 26 ›› Issue (5): 60-66.

• 基础研究 • 上一篇    下一篇

膜胆固醇氧化酶的萃取及特性研究

 李清, 贾洁, 牛天贵   

  1. 中国农业大学食品科学与营养工程学院
  • 出版日期:2005-05-15 发布日期:2011-09-19

Research on Extraction and Physical Properties of Membrane-bound Cholesterol Oxidase

 LI  Qing, JIA  Jie, NIU  Tian-Gui   

  1. College of Food Science and Nutritional Engineering, China Agricultural University
  • Online:2005-05-15 Published:2011-09-19

摘要: 胆固醇氧化酶在食品加工、医疗检测、生物抗虫方面的作用日益受到人们的重视,并且显示出巨大的应用潜力。本实验着重研究了膜结合型胆固醇氧化酶的萃取条件及其基本理化特性。10mmol/L磷酸钾缓冲液,pH7.5+0.5%TritonX-100的萃取液对胞膜酶的萃取效果最佳,萃取时间为10min。不含有TritonX-100的极性溶液的萃取效果较差,0.9%NaCl和pH7.5磷酸钾缓冲液相对而言具有稍好的萃取效果。酶活力测定体系中含有1%TritonX-100时显色最深,红色均匀且稳定持久,酶活力最高。酶蛋白中存在有含量相当高的糖组分,糖含量为3.763mg/ml,蛋白含量为1.106mg/ml,二者之比为3.4:1。酶反应的最适pH7.5,酶稳定范围pH5.0~1.0。粗酶的热稳定性较好,40℃以及40℃以下酶活力保持不变。

关键词: 膜胆固醇氧化酶, 萃取, Triton-100, 特性

Abstract: The significant functions of cholesterol oxidase in food processing and medical assay for blood cholesterol level and biological pest-resistance were now gradually recognized and showed great potential in application. In this paper, we investi- gated the extraction conditions and physical properties of the membrane-bound enzyme. The 10mmol/L potassium phosphate buffer containing 0.5%Triton X-100 of pH7.5 could efficiently extract the membrane-bound enzyme from the cells with the extraction time 10 minutes. The extraction effect of sodium chloride solution with the buffer without Triton X-100 was much worse than the buffer containing Triton X-100. The enzyme assay system containing 1% Triton X-100 showed the best color of red with good stability and the highest enzyme activity. The enzyme protein contained high level of saccharide, while the proportion of saccharide and protein was 3.4:1. The optimal pH for enzyme reaction was 7.5, and the enzyme remained stable within pH5.0 to 10.0. The enzyme had kept excellent thermal stability at temperature 40℃ and even lower than 40℃.

Key words: membrane-bound cholesterol oxidase, extraction, Triton X-100, physical properties