驯化对长焰煤制生物气过程氢化酶活性的响应分析

Response Analysis of Acclimation to Hydrogenase Activity During the Process of Biogenic Gas Generation from Long Flame Coal

以义马千秋矿长焰煤为产气底物,选取矿井水、沼液和污泥作为外加菌源,通过优化氢化酶活性的检测方法,并通过对菌源菌群的逐代驯化,考察其氢化酶活性与产气量及菌浊的响应关系。结果表明:氢化酶活性测试中以甘氨酸为电子载体的测定效果要优于以甲基紫精为电子载体的测定效果,而溶菌酶只对产氢菌氢化酶测试起作用。各菌源菌群驯化培养后氢化酶活性达到峰值的条件分别为:矿井水中产氢菌经三次驯化并培养48 h,峰值为0.11 mL/(mg·min);沼液中产氢菌经二次驯化并培养48 h,峰值为0.17 mL/(mg·min);污泥中产氢菌经二次驯化并培养56 h,峰值为0.14 mL/(mg·min);矿井水中产甲烷菌经三次驯化并培养56 h,峰值为2.84 mL/(mg·min);沼液中产甲烷菌经二次驯化并培养64 h,峰值为2.23 mL/(mg·min);污泥中产甲烷菌经二次驯化并培养80 h,峰值为2.45 mL/(mg·min);不同菌源菌群的产气效果和菌浊与各自驯化阶段表现出的氢化酶活性高度一致。

The hydrogen and methane production experiments were conducted with mine water,biogas slurry and sludge being selected as additional bacterial sources,and the long flame coal from Qianqiu Mine in Henan Province as the gas production substrate.The detection method of hydrogenase activity was optimized and the hydrogenase activity,the gas production and bacterial turbidity of different bacterial sources after the generational acclimation were investigated.The result show that the use glycine as the electron carrier performances better than the methyl viologen,while the lysozyme only works on the hydrogenase test of the hydrogen-producing bacteria.The conditions for the hydrogenase activity of each bacterial source acclimation and culture to reach the peak are as follows:under the third-time acclimation and culture for 48 h,the peak value for the hydrogen-producing bacteria in the mine water corresponds to 0.11 mL/(mg·min);under the secondary acclimation and culture for 48 h,the peak value reaches 0.17 mL/(mg·min)for the biogas slurry;under the secondary acclimation and culture for 56 h,the peak value reaches 0.14 mL/(mg·min)for the sludge;under the third-time acclimation and culture for 56 h,the peak value corresponds to 2.84 mL/(mg·min)for the methanogenic bacteria in the mine water;under the secondary acclimation and culture for 64 h,the peak value reaches 2.23 mL/(mg·min)for the biogas slurry;and under the secondary acclimation and culture for 80 h,the peak value for the sludge reaches 2.45 mL/(mg·min).The gas production effect and bacterial turbidity of different bacterial sources are highly consistent with the hydrogenase activity shown in the respective acclimation stages.