裂褶菌F17锰过氧化物酶酶活力影响因素的响应面优化 (英文)
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安徽省自然科学基金项目 (No. 2010) 资助。


Optimization of process variables for the manganese peroxidase of the white-rot fungus Schizophyllum sp. F17 by full factorial central composite design
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Natural Science Research Program of the Educational Of?ce of Anhui Province (No. 2010).

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    摘要:

    锰过氧化物酶是真菌分泌的一种糖基化的含有血红素辅基的胞外蛋白, 在染料降解和脱色过程中起着重要作用。本实验利用本实验室保存的的白腐真菌裂褶菌Schizophyllum sp. F17产锰过氧化物酶(MnP),研究MnP的酶学性质,并对酶活条件进行优化。实验通过超滤浓缩、DEAE-纤维素、DE52离子交换层析和Sephadex G-75凝胶过滤等步骤,分离纯化得到电泳纯的锰过氧化物酶。该酶蛋白含量为23 μg/mL,分子量大小为49.2 kDa,在0.1 mmol/L H2O2中半衰期为5~6 min。Mn2+、H2O2以及酶的用量可以影响MnP酶促反应的效率,在单因子分析法的基础上,通过全因子中心组合设计响应面分析表明:H2O2以及H2O2与酶用量之间的交互作用对酶促反应的作用是最显著的。在优化条件下,酶对偶氮染料金橙G、刚果红显示出较强的脱色能力。

    Abstract:

    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.

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查诚,荚荣,陶香林,姚祖亮. 裂褶菌F17锰过氧化物酶酶活力影响因素的响应面优化 (英文)[J]. 生物工程学报, 2010, 26(3): 341-349

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  • 收稿日期:2009-09-07
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