植物血红素加氧酶活性的微量比色测定法与盐胁迫诱导特性的初步研究.PDF

南京农业大学学报摇 2009,32 (3): 146 150 摇 Journal of NanjingAgricultural University http:/ / nauxb郾njau郾edu郾cn 陈舟舟, 齐继艳, 张亮, 等. 植物血红素加氧酶活性的微量比色测定法与盐胁迫诱导特性的初步研究 [J]. 南京农业大学学报, 2009, 32(3): 146 150 植物血红素加氧酶活性的微量比色测定法与 盐胁迫诱导特性的初步研究 陈舟舟, 齐继艳, 张亮, 徐晟, 沈文飚* (南京农业大学生命科学学院/ 生命科学实验中心, 江苏 南京 210095) 摘要: 建立并改进了植物血红素加氧酶 (heme oxygenase, HO) 活性的微量比色测定法。 测定体系的最佳反应条件为: 最适pH7郾2, 最适温度37 益, 加入5~20 滋L酶液量 (总蛋白量控制在5~10 滋g), 酶促反应时间为60 min。 并通过比 较正常生长和盐胁迫条件下多种植物的HO活性变化, 初步证明HO具有盐胁迫诱导特性。 关键词: 酶活性测定; 血红素加氧酶; 盐胁迫诱导特性 中图分类号: Q5 33摇 摇 摇 文献标志码: A摇 摇 摇 文章编号: 1000 2030 (2009) 03 0146 05 A micro鄄volume spectrometry for determination of heme oxygenase activity in plants and preliminary research on its salt鄄inducible character CHEN Zhou鄄zhou, QI Ji鄄yan, ZHANG Liang, XU Sheng, SHEN Wen鄄biao* (College of Life Sciences/ Laboratory Centre of Life Sciences, Nanjing Agricultural University, Nanjing210095, China) Abstract: A micro鄄volume spectrometry for determination of heme oxygenase (HO) activity in plants was established and further modified. The optimum conditionsfor determination were as follows: the optimum pH and temperature were 7郾2 and 37 益, the applied volume of extractwasabout5 20 滋L (equivalent to5 10 滋gprotein), and60 minwaschosen asreactiontime. Final鄄 ly, in comparison with the changes of HO activities under normal growth condition or salt stressin several plant speciesasreaction time, we also preliminarily discovered that HO exhibits the salt鄄inducible character. Key words:determining enzyme activity; heme oxygenase; salt鄄inducible character 2+ 一氧化碳 (carbon monoxide, CO)、 胆绿素 (biliverdin 御, BV) 和游离二价铁离子 (Fe ) 是血 红素加氧酶 (heme oxygenase, HO, EC 1郾14郾99郾3) 催化血红