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Purification and characterization of an extracellular laccase from the anthracene-degrading fungus Fusarium solani MAS2

机译:蒽降解镰刀菌MAS2胞外漆酶的纯化和鉴定

摘要

An extracellular laccase was purified from the culture medium of the non-white rot, anthracene-degrading fungal strain Fusarium solani MAS2. Both native PAGE and SDS-PAGE revealed one single band corresponding to a molecular weight of about 72 kDa. Treatment with endoglycosidase H reduced the molecular weight by 12%. The purified laccase maintained stable at pH 3-11 and up to 50 degrees C. The highest activity was detected at pH 3.0 and at 70 degrees C. The enzyme retained 46.2-97.2% of it activity in the presence of 20 mM Pb(2+), Ni(2+), Cr(3+), and its activity was enhanced in the presence of 20 mM Hg(2+). The laccase retained more than 50% of its activity in the presence of 5% acetone, acetonitrile, dimethyl sulphoxide (DMSO), ethanol and methanol. The kinetic constants (K(m) and K(cat)) showed that 2,6-dimethoxyphenol (DMOP) and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) diammonium salt (ABTS) were the more effective substrates rather than catechol and guaiacol. The novel properties of this laccase suggest its potential for biotechnological and environmental applications. (C) 2010 Elsevier Ltd. All rights reserved.
机译:从非白色腐烂,蒽降解真菌菌株Fusarium solani MAS2的培养基中纯化细胞外漆酶。天然PAGE和SDS-PAGE均显示出一条对应于约72kDa分子量的单条带。用糖苷内切酶H处理使分子量降低了12%。纯化的漆酶在pH 3-11和高达50摄氏度的温度下保持稳定。在pH 3.0和70摄氏度的条件下检测到最高的活性。在20 mM Pb(2)存在的情况下,该酶保留了其46.2-97.2%的活性。 +),Ni(2 +),Cr(3+)及其活性在存在20 mM Hg(2+)的情况下得到增强。在5%丙酮,乙腈,二甲亚砜(DMSO),乙醇和甲醇的存在下,漆酶保留了超过50%的活性。动力学常数(K(m)和K(cat))显示2,6-二甲氧基苯酚(DMOP)和2,2'-叠氮基双-(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)为比邻苯二酚和愈创木酚更有效的底物。这种漆酶的新颖特性表明其在生物技术和环境应用中的潜力。 (C)2010 Elsevier Ltd.保留所有权利。

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