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Construction of Recombinant E.coli Expressing A Thermophilic Catalase and Its Application in H_2O_2 Wastewater Treatment

机译:表达嗜热性过氧化氢酶的重组大肠杆菌的构建及其在H_2O_2废水处理中的应用

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The recombinant plasmid pETac-CATHis with tac promoter was constructed for the constitutive expression of a thermophilic catalase in E.coli. And the effects of cultivation conditions of two recombinant strains, BL21(DE3)/pETac-CATHis and BL21(DE3)pLysS/pETac-CATHis, such as initial pH, cultivation temperature, cultivation time, and loading volume on catalase activity were investigated. The optimal cultivation conditions were: the initial pH 7.0, cultivation temperature 37°C, cultivation time 12 hours, and loading volume 25 mL in 100 mL flask. Under the optimal cultivation condition, the highest catalase activities of the two recombinant strains were 291.7 U/mL and 349.5 U/mL, respectively. After adding catalase, H_2O_2 in simulation H_2O_2 wastewater containing phenol was decomposed completely very quickly. Declining of catalase activity was detected with enhancing the phenol concentration, probably because the phenol might be a kind of competitive inhibitor for catalase.
机译:具有TAC启动子的重组质粒pETAC-阴离子被构建用于大肠杆菌中嗜热性过氧化氢酶的组成型表达。研究了两种重组菌株的培养条件,BL21(DE3)/ PETAC-阴离子和BL21(DE3)帘布层/ PETAC-阴性的影响,例如初始pH,培养温度,培养时间和在过氧化度酶活性上加载量。最佳培养条件是:初始pH 7.0,培养温度37℃,培养时间12小时,并在100mL烧瓶中加载25ml。在最佳培养条件下,两种重组菌株的最高过氧化酶活性分别为291.7U / mL和349.5U / mL。在加入过氧化氢酶后,在仿真H_2O_2含有苯酚的废水完全非常快速地分解。通过增强酚浓度检测到过氧化氢酶活性的下降,可能是苯酚可能是过氧化氢酶的竞争性抑制剂。

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