彩绒革盖菌CV-8胞外漆酶的诱导、纯化及部分性质研究

Studies on Induce Condition,Purification and Partial Character of Extracellular Laccase from Coriolus Versicolor CV-8

为探讨彩绒革盖菌(Coriolus versicolor)漆酶的生产条件和性质,对其进行了硫酸铵盐析、半透膜透析、DEAE-纤维素离子交换、Sephadex G100柱层析纯化,粗酶液被纯化36.1倍,比活力10180u/mg,回收率44.6%。凝胶过滤法测定酶的分子量为62kDa。研究了漆酶的产酶曲线及酶作用最适条件。结果表明在该培养条件下,彩绒革盖菌第11d达产酶高峰,峰值酶活为398u/ml,酶作用的最适pH值为4.6,最适温度为25℃,Mg2+、Mn2+、Cu2+对漆酶有激活作用,而Ag+、Fe3+和Cl-则抑制漆酶活性。以邻联甲苯胺为底物的表观Km值为892×10-3mol/L。该菌可作为木质素降解和环保工业漆酶生产菌种。

The laccase of Coriolus versicolor CV - 8 was purified by salting out from ammonium sulfate, dialyzed by semipermeable membrane, DE- AE - Cellulose ion exchange and Sephadex G100 column chromatography in order to explore its productive situation and characteristics. It was purified 36.1 times; and the specific activity and recovery of purification laccase were 10180 u/mg and 46.6% respectively.The curve of laccase production and optimal condation of laccase reaction were also studied. The results showed that Coriolus versicolor came the laccase production peak which was 398u/ml at the 11 day in this culture condition. The optimal temperature and optimal pH of laccase reaction were at 25℃ and pH4.6. Mg^2 ＋ , Mn^2 ＋ and Cu^2 ＋ increased laccase activity whereas Ag^ ＋ , Fe^3 ＋ and Cl^- obviously inhibited it. The form view Km of laccase was 892 × 10^- 3 mol/L with o - Tolidine as substrate. This fungus can be the productive strain of the Lignin Degradation and the laccase in envirmental protection industry.