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杂色云芝组成型漆酶Ⅰ的纯化和底物专一性
Purification and Substrate Specificity of a Constitutive Laccase Ⅰ from a White-rot Fungus Trametes versicolor
【摘要】 采用合成培养基培养杂色云芝As5 4 8,从发酵液中纯化出一种组成型漆酶同功酶Ⅰ .经超滤浓缩 ,DEAE SephadexA 5 0离子交换层析 ,Bio gelP 10 0凝胶过滤纯化了该酶 .SDS PAGE分析发现 ,该酶分子量为 6 8kD ,薄层等电聚焦测得等电点为 3 5 .漆酶Ⅰ的底物范围较宽 ,以O2 为电子受体 ,可以氧化多种木素单体模型物 ,包括 2 ,6 二甲氧基酚 ,2 ,2′ 联氮 二 (3 乙基 苯并噻唑 6 磺酸 )(ABTS) ,愈创木酚 ,咖啡酸 ,阿魏酸和邻联茴香胺 .结果表明 ,该酶在木质素的生物降解中可能有重要的作用和应用价值 .
【Abstract】 A constitutive laccase Ⅰ (EC 1.10.3.2) isolated from Trametes vesicolor As5.48 was purified to electrophoretic homogeneity by ultrafiltration, DEAE Sephadex A50 ion exchange chromatography and gel filtration on Bio gel P 100. The molecular weight of the enzyme was 68 000 determined by SDS PAGE and the isoelectric point was estimated to be pH 3 5 by IEF. The enzyme has a wide range of substrates in the presence of oxygen as the electron receptor, including 2,6 dimethoxyphenol, ABTS, guaiacol, caffeic acid, ferulic acid, o dianisidine.The results indicates that the enzyme may play an important role in lignin degradation.
【Key words】 Trametes versicolor; laccase; purification; lignin model compound;
- 【文献出处】 中国生物化学与分子生物学报 ,Chinese Journal of Biochemistry and Molecular Biology , 编辑部邮箱 ,2002年06期
- 【分类号】Q554
- 【被引频次】24
- 【下载频次】228