斜卧青霉菌P6木素过氧化物酶的纯化与特性

Purification and identification of lignin peroxidase from Pencillium documbens P6

木素过氧化物酶是降解木素类物质的主要酶类。使用硫酸铵盐析、DEAE 纤维素和CM 纤维素离子交换层析、SephadexG 10 0凝胶过滤等分离与纯化技术 ,从斜卧青霉菌P6 (PenicilliumdocumbensP6 )发酵培养 7d的液体培养液中分离纯化得到电泳纯的木素过氧化物酶 ,经SDS PAGE鉴定为单一条带 ,表观相对分子质量为 4 6 3× 10 3 。以藜芦醇为底物测定了该酶的Km,vmax,作用的 pH和温度范围 ,结果为Km0 5 6 5mmol·L-1,vmax0 0 88mmol·(L·min) -1,pH 4～ 9,温度 2 5～ 5 5℃。N末端序列为VLLPADEKNA ,与真菌LiP无同源性。

Lignin peroxidase was separated and purified from a liquid 7 day culture filtrate of Pencillium documbens P6 by ammonium sulfate precipitation, DEAE cellulose and CM cellulose ion exchanger resin chromatography, Sephadex G 100 gel filtration chromatography. There was a single band in SDS PAGE and molecular weight was estimated to be 46 3×10 3 by SDS PAGE. K m is 0 565?mmol·L -1 , v max is 0 088?mmol·(L·min) -1 . The effect of pH and temperature on enzyme activity using veretryl alcohol as substrate were determined. The pH range is 4-9 and the temperature range is 25?℃-55?℃ The N terminal amino acid residue was VLLPADEKNA, there was no homologous with other fungal LiP.