Zha Cheng, Jia Rong, Tao Xianglin, Yao Zuliang
School of Life Science, Anhui Key Laboratory of Eco-engineering and Bio-technique, Anhui University, Hefei 230039, China.
Sheng Wu Gong Cheng Xue Bao. 2010 Mar;26(3):341-9.
White-rot fungus manganese peroxidase (MnP) that has great potential in degrading azo dyes is one of the extracellular glycolsylated heme proteins. MnP from Schizophyllum sp. F17 was isolated and purified by Sephadex G-75 gel filtration chromatography followed by DEAE-cellulose anion exchange chromatography. The molecular weight of the puried enzyme was 49.2 kDa, while the half-life of the MnP in the presence of 0.1 mmol/L H2O2 was 5-6 min. The efficiency of MnP-catalyzed reactions were determined by three key factors: the concentrations of Mn2+, H2O2, and the amount of MnP. Using single factor analysis, an optimized concentration of Mn2+, H2O2 and enzyme were optimized to be 1.2 mmol/L, 0.1 mmol/L, and 0.4 mL, respectively. A response surface methodology (RSM) employing two-level-three-factor full factorial central composite design was used to optimize the catalytic conditions. The result showed that the concentration of H2O2 and the interaction between H2O2 and MnP mostly affect the MnP catalytic efficiency. Finally, we show that the azo dyes could be efficiently decolorized by the purified MnP under optimized conditions.
白腐真菌锰过氧化物酶（MnP）是一种细胞外糖基化血红素蛋白，在降解偶氮染料方面具有巨大潜力。通过Sephadex G - 75凝胶过滤色谱法，随后进行DEAE - 纤维素阴离子交换色谱法，从裂褶菌属F17中分离并纯化了MnP。纯化后的酶分子量为49.2 kDa，而在0.1 mmol/L H2O2存在下，MnP的半衰期为5 - 6分钟。MnP催化反应的效率由三个关键因素决定：Mn2 +、H2O2的浓度以及MnP的量。通过单因素分析，将Mn2 +、H2O2和酶的优化浓度分别优化为1.2 mmol/L、0.1 mmol/L和0.4 mL。采用二级三因素全因子中心复合设计的响应面法（RSM）来优化催化条件。结果表明，H2O2的浓度以及H2O2与MnP之间的相互作用对MnP催化效率影响最大。最后，我们表明在优化条件下，纯化的MnP可以有效地使偶氮染料脱色。
