凹凸棒石粘土固定化辣根过氧化物酶处理废水研究
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摘要
本文以凹凸棒石粘土(简称“凹土”)为原料,水玻璃为绞结剂,淀粉为造孔剂,研究制备了用于辣根过氧化物酶固载的复合凹土,并对影响载体散失率的各种因素进行正交实验,确定了最佳载体制备条件;通过单因素平行实验,确定了固定化HRP的最佳条件、降解苯胺与硝基酚的最佳条件以及固定化HRP的重复使用性能;通过Lineweaver-Burk线性拟合初始硝基酚浓度与反应速度的关系,确定了固定化HRP降解硝基酚的动力学方程、最大反应速度Vmax以及米氏常数Km。
实验结果表明:凹土与水玻璃和淀粉的质量比为100:12:20,造粒于550℃马弗炉中煅烧时间2 h,此时固定HRP的载体具备高机械强度;固载的工艺条件为常温,固定时间1.5 h,单位活度载体量1.5 mg/U,pH值5.0;固定化酶降解苯胺废水反应时,反应时间为1.5小时,不需要缓冲体系控制pH值,每毫升100mg/L的苯胺废水需要4.8U的固定酶,H2O2与苯胺的摩尔比为1.2:1;在此条件下固定化HRP可以重复使用4次,并均能维持90%以上的苯胺去除率。固定化HRP处理硝基酚废水的最佳条件为反应时间1.5 h,每毫升100 mg/L的硝基酚废水需6.0 U的给酶活(按固定率为95%计算,固定酶活为5.7 U),H2O2与硝基酚的最佳化学计量比为1.2:1;固定化HRP具有较好的重复使用性能,经重复使用7次后,硝基酚的去除率仍大于60%;固定化HRP降解硝基酚的动力学方程为:1/v = 1.403/[S] - 0.081,最大反应速度Vmax = 12.35 mg/L·min,米氏常数Km = 17.32 mg/L。
Attapulgite clay immobilized horseradish peroxidase was successfully prepared,the effects of the vector formulation、immobilization conditions and reaction conditions are studied, the optimal conditions for the preparation of the carrier, enzyme immobilization and reaction conditions are determined through the orthogonal experiments. The optimal conditions for HRP immobilization, nitrophenol degradation reaction, the repeatability of immobilized HRP are also investigated through single factor parallel experiments. the kinetic parameter of degradation of aniline and nitrophenol by immobilized HRP, the maximum response velocity and michaelis constant (Km) by Lineweaver-Burk linear fitting between the initial concentration of nitrophenol and response velocity are investigated.
The results shows when the amount of mass ratio of attapulgite clay : sodium silicate: soluble starch are 100:12:20,the calcination temperature of 550℃and the calcination time of 2 h, we can get carriers holding the best absorption properties. The optimal technological parameters for HRP immobilization are PH = 5.0,the volume of unit vector activity of the 1.5 mg/U and the immobilized 1.5 h at room temperature.When non-adjust PH, the reaction time is 1.5 h, 100 mg/L of aniline need 4.8 U enzyme activity, the molar ratio is H_2O_2 and aniline is 1.2, the immobilized HRP can be reused for 4 times and aniline degradation efficiency maintain over 90%.
In the meantime, when non-adjust PH, the reaction time is 1.5 h, 100 mg/L of nitrophenol need 6.0 U enzyme activity, the molar ratio is H2O2 and nitrophenol is 1.2,t he immobilized HRP can be reused for 7 times and nitrophenol degradation efficiency maintain over 60%.The kinetic equation is 1/v = 1.403/[S] - 0.081, the Vmax and Km are 12.35 mg/L·min,and 17.32 mg/L, respectively.
实验结果表明:凹土与水玻璃和淀粉的质量比为100:12:20,造粒于550℃马弗炉中煅烧时间2 h,此时固定HRP的载体具备高机械强度;固载的工艺条件为常温,固定时间1.5 h,单位活度载体量1.5 mg/U,pH值5.0;固定化酶降解苯胺废水反应时,反应时间为1.5小时,不需要缓冲体系控制pH值,每毫升100mg/L的苯胺废水需要4.8U的固定酶,H2O2与苯胺的摩尔比为1.2:1;在此条件下固定化HRP可以重复使用4次,并均能维持90%以上的苯胺去除率。固定化HRP处理硝基酚废水的最佳条件为反应时间1.5 h,每毫升100 mg/L的硝基酚废水需6.0 U的给酶活(按固定率为95%计算,固定酶活为5.7 U),H2O2与硝基酚的最佳化学计量比为1.2:1;固定化HRP具有较好的重复使用性能,经重复使用7次后,硝基酚的去除率仍大于60%;固定化HRP降解硝基酚的动力学方程为:1/v = 1.403/[S] - 0.081,最大反应速度Vmax = 12.35 mg/L·min,米氏常数Km = 17.32 mg/L。
Attapulgite clay immobilized horseradish peroxidase was successfully prepared,the effects of the vector formulation、immobilization conditions and reaction conditions are studied, the optimal conditions for the preparation of the carrier, enzyme immobilization and reaction conditions are determined through the orthogonal experiments. The optimal conditions for HRP immobilization, nitrophenol degradation reaction, the repeatability of immobilized HRP are also investigated through single factor parallel experiments. the kinetic parameter of degradation of aniline and nitrophenol by immobilized HRP, the maximum response velocity and michaelis constant (Km) by Lineweaver-Burk linear fitting between the initial concentration of nitrophenol and response velocity are investigated.
The results shows when the amount of mass ratio of attapulgite clay : sodium silicate: soluble starch are 100:12:20,the calcination temperature of 550℃and the calcination time of 2 h, we can get carriers holding the best absorption properties. The optimal technological parameters for HRP immobilization are PH = 5.0,the volume of unit vector activity of the 1.5 mg/U and the immobilized 1.5 h at room temperature.When non-adjust PH, the reaction time is 1.5 h, 100 mg/L of aniline need 4.8 U enzyme activity, the molar ratio is H_2O_2 and aniline is 1.2, the immobilized HRP can be reused for 4 times and aniline degradation efficiency maintain over 90%.
In the meantime, when non-adjust PH, the reaction time is 1.5 h, 100 mg/L of nitrophenol need 6.0 U enzyme activity, the molar ratio is H2O2 and nitrophenol is 1.2,t he immobilized HRP can be reused for 7 times and nitrophenol degradation efficiency maintain over 60%.The kinetic equation is 1/v = 1.403/[S] - 0.081, the Vmax and Km are 12.35 mg/L·min,and 17.32 mg/L, respectively.
引文
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