凤尾菇漆酶性质及应用的研究

The studies on characterization and Application of edible fungi-Pleurotus eryngii

液体发酵培养凤尾菇Pleurotus eryngii（凤杰一号）,并对该菌株进行漆酶酶学性质和染料脱色的研究。粗酶液经盐析、透析、浓缩初步纯化后,漆酶活力达4.27 U.mL^-1。该酶催化ABTS氧化反应的最适pH为4.5,温度为65℃,在60℃以下、pH 4.5-6.0该酶的稳定性最好,以ABTS为底物的km为0.335 mmo.lL^-1,Vmax为0.795μmol.^L-1.min^-1,反应时间4-5 min为佳。该酶在不同底物的作用下,ABTS的氧化能力最强,愈创木酚、氢醌、儿茶酚次之,阿魏酸无氧化能力。叠氮化钠是该漆酶高效抑制剂,浓度为0.5 mmol.L^-1使漆酶活性完全抑制。该酶对三苯甲烷染料（孔雀绿、结晶紫）均有降解脱色性能,脱色率分别达到72.3%和50%。但该酶对孔雀绿的脱色能力比结晶紫要好,同时在体系中加入ABTS可提高脱色能力。

In the research, By fermentation of Pleurotus eryngii （Fengjie NO 1）, and Futher studies on the characterization and decolor ability were carried out with this strain. With preliminary purification by salting out, dialyzing and condensing, the enzyme activity reached 4.27 U mL^-1. The enzyme is stable below 60 ℃ and in the pH range of 4.5-6.0. The optimum pH and temperature for oxidation of ABTS were 4.5 and 65 ℃, respectively. The km value is 0. 335 mmol · L^-1 and Vmax is 0. 795μmol · L ^-1. min^-1 with ABTS as the substrate. The reaction time is 4-5 min. Under different substrates, ABTS displayed maximum oxygenation capability, Guaiacol, Hydroquinone, catechol take second place, Ferulic acid have no oxygenation capability. Sodium azide is the most efficient inhibitor, which can restrain laccase activity completely at 0.5 mmol · L^-1. The laccase could decolor triphenylmethane （mal achite green stain, crystal violet） . And from the research, The laccase decoloring malachite green stain is better than crystal violet. Moreover, adding ABTS into the system could impove its discoloring ability.