白腐真菌SYBC-L2漆酶的分离纯化及其在毛纺染料脱色中的应用  

Purification of Laccase from the White Rot Fungus Strain SYBC-L2 and Its Application in Decolorization of Woolen Dyes

作　　者：王志新[1] 施晓燕[1] 蔡宇杰[1] 廖祥儒[1] 

机构地区：[1]江南大学工业生物技术教育部重点实验室,无锡214122

出　　处：《生物技术通报》2009年第8期156-161,165,共7页Biotechnology Bulletin

基　　金：长江学者和创新团队发展计划(IRT0532)

摘　　要：经过硫酸铵分级盐析、DEAE-cellulose离子交换层析、SephacrylTMS-200柱层析,从白腐菌株Phellinus sp.SYBC-L2所产的漆酶中纯化得到电泳纯漆酶Lac3,其纯化倍数为28.27,回收率为14.76%。Lac3为一种糖蛋白,糖含量29.66%,SDS-PAGE表观分子量约为64.3kD;其催化氧化DMP(2,6-Dimethoxyphenol)的表观Km值和Vmax分别为1.01mmol/L和186.2U/mg蛋白。Lac3的最适温度为60℃,最适pH为3.5;在30℃～50℃和pH4.5～9.0范围内稳定。Cu2+、SO42-、CO32-对Lac3具有一定促进作用,而Fe3+、Fe2+、NO3-则具有抑制作用。色素的最佳脱色条件为(Lac3终浓度为2U/ml):媒介元PV在温度50℃、pH值3.0的条件下反应2h,脱色率可达95.10%;弱酸蓝在温度40℃、pH值4.0的条件下作用3h,脱色率为75.09%。The laccase Lac3 from Phellinus sp. SYBC-L2 was purified by salt precipitation with ammonium sulphate, DEAE-cellulose ion exchange column chromatography and SephacrylTMS-200 gel column chromatography. Purification of 28.27 fold was achieved with an overall yield of 14.76 %. Lac3 was a glycnprotein with carbohydrate content of 29.66%. SDS-PAGE showed that the apparent molecular weight of the laccase was 64.3 kD. The apparent Km and Vmax of Lac3 for DMP was 1.01 mmol/L and 186.2 U/mg proteins, respectively. The optimum temperature and pH value was 60℃ and 3.5, respectively. Lac3 was stable within 30℃ to 50℃ , and pH range from 4.5 to 9.0. The laccase activity was enhanced by Cu2＋ , SO42- and CO32- , while inhibited by Fe3＋ , Fe2＋ and NO3-. The optimal condition was obtained as follows, the decolorization rate of Diamond Black PV reached 95.10 % in 2 h at 50℃ , pH 3.0, and the decolorization rate of disperse black reached 75.09% in 3 h at 40℃ , pH 4.0.

关 键 词：PHELLINUS sp SYBC-L2 漆酶 分离纯化 酶学性质 脱色 

分 类 号：X791[环境科学与工程—环境工程]