Purification and characterization of laccases from the white-rot basidiomycete Dichomitus squalens.
Purification and characterization of laccases from the white-rot basidiomycete Dichomitus squalens.
复制标题
白腐担子菌 Dichomitus squalens 漆酶的纯化和表征。
DOI：
10.1006/abbi.1998.0625
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Gold,MH
中科院分区：
文献类型：
--
作者：
Perie,FH;Reddy,GV;Blackburn,NJ;Gold,MH
用DEAE-Sepharose和Mono-Q快速蛋白液层析法，从角点毛滴虫木质素分解培养物的胞外培养液中分离纯化了两种漆酶c1和c2，纯化倍数约为225倍。经SDS-PAGE测定，各均一漆酶(c1和c2)的相对分子质量约为66 kDa。这两种形式都是糖蛋白，每种形式的蛋白质分子都含有四个铜原子。这两种漆酶的N-末端序列的前20个氨基酸是相同的，并且与其他木质素降解真菌的漆酶相似。这些漆酶的电子吸收光谱在610和330 nm处有吸收带，表明它们是第一类和第三类铜。漆酶C1的EPR谱显示出指示I型和II型铜的条带。该漆酶能氧化多种酚类底物，其氧化2，6-二甲氧基苯酚的最适pH为3.0，并受氟化物和叠氮化物的强烈抑制。
Two chromatographic forms of laccase c1 and c2 were purified approximately 225-fold from the extracellular culture fluid of ligninolytic cultures ofDichomitus squalens,using DEAE–Sepharose and Mono-Q fast protein liquid chromatography. Each homogeneous laccase (c1 and c2) has a molecular mass of approximately 66 kDa as determined by SDS–PAGE. Both forms are glycoproteins, and each contains four copper atoms per molecule of protein. The first 20 amino acids of the N-terminal sequences of these two laccases are identical and are similar to those of laccases from other lignin-degrading fungi. The electronic absorption spectra of these laccases exhibit bands at 610 and 330 nm, indicative of type I and type III copper. The EPR spectrum of laccase c1 exhibits bands indicative of type I and type II copper. Each laccase oxidizes a variety of phenolic substrates, has a pH optimum of 3.0 for the oxidation of 2,6-dimethoxyphenol, and is inhibited strongly by fluoride and azide.