初级基质对降解菌Pseudomonas putida共代谢苯扎贝特的影响

Effect of primary substrate on co-metabolism of bezafibrate by Pseudomonas putida

通过对3,5-二硝基水杨酸(DNS)法测定还原糖含量各影响因素的筛选,优化确定了显色时间8 min,稳定时间10 min,最佳波长490 nm下进行吸光度测定的检测方法;分析分别以葡萄糖和麦芽糖作为外加碳源时,降解菌Pseudomonas putida B-31的生长情况和共代谢降解典型药物苯扎贝特(BZF)的过程。结果表明,降解菌只有在外加碳源的条件下才可正常生长,而且其在葡萄糖环境中生长得更好;拟合得到的葡萄糖、麦芽糖和BZF代谢动力学结果显示,葡萄糖对BZF去除的促进作用更为明显,同时从葡萄糖培养基中降解菌所提取的酶比活力要高于麦芽糖培养基,分析原因可能是葡萄糖所诱导的降解菌关键酶活力更强,而且还可能会产生不同的蛋白质点位。

Factors affecting the detection of reducing sugars using 3,5-dinitrosalicylic acid （DNS） method were screened. Also,the method was optimized as follows：color-development time of 8 min, stabilization time of 10 min and detection wavelength of 490 nm. Growth of the degrading strains of Pseudornonas putida B-31 and its co-metabolism of bezafibrate were investigated by adding glucose and/or maltose as the primary substrate. The re- sults showed that the degrading strains could only grow in the presence of glucose or maltose, and better growth was obtained with glucose. The fitting results of the metabolic kinetics of glucose,maltose and bezafibrate indica- ted that glucose could evidently accelerate the degradation of bezafibrate. Additionally,the specific activity of en- zymes extracted from the degrading strains growing in the glucose culture media was higher than that in the malt- ose culture media. The possible reason was that the activity of enzymes induced by glucose was much higher and different protein spots were generated.