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首页> 外文期刊>Biochemistry >Isolation and Purification of Enzymes from Ligninolytic Complex of the Basidial Fungus Trametes pubescens (Schumach.) Pilat and Study of Their Properties
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Isolation and Purification of Enzymes from Ligninolytic Complex of the Basidial Fungus Trametes pubescens (Schumach.) Pilat and Study of Their Properties

机译：毛霉菌（Schumach。）Pilat木质素分解复合物酶的分离纯化及其性质的研究

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摘要

A method for purification of enzymes from the ligninolityc complex of the basidiomycete Trametes pubescens (Schumach.) Pilat has been elaborated.Two homogeneous isoforms of laccases (laccase 1 and laccase 2) as well as a homogeneous preparation of lignin peroxidase were isolated.Basic biochemical parameters of the enzymes were determined,such as the molecular weights (67,67,and 45 kD,respectively),isoelectric points (5.3,5.1,and 4.2,respectively),as well as content and composition of the carbohydrate moiety of the laccases (N-acetylglucosamine,mannose,and xylose).The pH dependences and thermal stabilities of the laccases were investigated.The kinetic parameters of the enzymatic reactions catalyzed by the laccases were determined using different substrates,such as catechol,hydroquinone,2,2'-azinobis-(3-ethyl-benzthiazoline-6-sulfonate),and K_4Fe(CN)_6.The structure of the active sites of both laccases and the lignin peroxidase were studied by EPR,CD,and UV-VIS spectroscopy,as well as using fluorescence analysis.Our studies showed similarity of the spectral characteristics of the two laccases,whereas their kinetic properties were found to be different.
机译：提出了一种从担子菌Trametes pubescens（Schumach。）Pilat的木质素醇复合物中纯化酶的方法，分离了漆酶的两种同质同工型（漆酶1和漆酶2）以及木素过氧化物酶的均质制剂。确定了酶的参数，例如分子量（分别为67、67和45 kD），等电点（分别为5.3、5.1和4.2）以及漆酶的碳水化合物部分的含量和组成（N-乙酰氨基葡萄糖，甘露糖和木糖）。研究了漆酶的pH依赖性和热稳定性。使用不同的底物，例如邻苯二酚，对苯二酚，2,2'，确定了漆酶催化的酶反应动力学参数。 -氮杂双-（3-乙基-苯并噻唑啉-6-磺酸盐）和K_4Fe（CN）_6。通过EPR，CD和UV-VIS光谱研究了漆酶和木质素过氧化物酶的活性位点结构华硕我们的研究表明两种漆酶的光谱特性相似，但发现它们的动力学性质不同。

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