白腐真菌落叶松锈迷孔菌产漆酶液体培养基的优化及其对染料的脱色作用

以白腐真菌落叶松锈迷孔菌(Porodaedalea laricis)胞外漆酶为响应值,通过将Plackett-Burman设计、最陡爬坡设计和Box-Behnken设计相结合,获得了P.laricis产胞外漆酶的最适培养基为:去皮马铃薯365.61 g/L、蛋白胨5.0 g/L、葡萄糖20.0 g/L、KH2PO41.0 g/L、MgSO47H2O 0.5 g/L、MnSO4 H2O 0.15 g/L、CaCl22H2O 0.03 g/L、酒石酸铵6.68 g/L、琥珀酸钠1.5 g/L、吐温800.48 mL/L、玉米芯46.43 g/L、维生素B10.01 g/L。在该条件下,P.laricis漆酶活性为3.29 U/mL,相比于优化前提高了2.81倍,与理论值3.32 U/mL相近,说明该模型准确可靠。此外,将漆酶应用于降解多种合成染料包括活性亮蓝X-BR、雷马素亮蓝R、酸性黑172、刚果红、亚甲基蓝、中性红、靛蓝、萘酚绿B和结晶紫,反应168 h后脱色率分别可达到95.64%、97.21%、36.11%、91.63%、61.42%、74.65%、48.60%、25.13%和68.80%。

Medium Optimization for the Laccase Production by White Rot Fungus Porodaedalea laricis and Its Dye Decolorizing Capacity

In this study,Plackett-Burman design,path of the steepest ascent,and Box-Behnken design were used to optimize the liquid medium composition for laccase production of white rot fungus Porodaedalea laricis with the laccase activity set as a response.The optimal compositions for production of P.laricis laccase was obtained as follows:peeled potato 365.61 g/L,peptone 5.0 g/L,glucose 20.0 g/L,KH2PO41.0 g/L,MgSO47H2O 0.5 g/L,MnSO4 H2O 0.15 g/L,CaCl22H2O 0.03 g/L,ammonium tartrate 6.68 g/L,sodium succinate 1.5 g/L,Tween 800.48 mL/L,corncob 46.43 g/L,and vitamin B10.01 g/L.Under the optimized conditions,the laccase activity was 3.29 U/mL,which was 2.81-fold of that given in the basal medium,and this yield was very in good agreement with the theoretical result of 3.32 U/mL,indicating the model was accurate and reliable.Moreover,the laccase exerted notably decolorizing capacity for many synthetic dyes including Reactive Brilliant Blue X-BR,Remazol Brilliant Blue R,Acid Black 172,Congo Red,Methylene Blue,Neutral Red,Indigo Blue,Naphthol Green B,and Crystal Violet,with the respective decolorization rate of 95.64%,97.21%,36.11%,91.63%,61.42%,74.65%,48.60%,25.13%,and 68.80%after 168 h.