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Demonstration of Laccase in the White Rot Basidiomycete Phanerochaete chrysosporium BKM-F1767

机译:在白色腐烂担子菌Phanerochaete chrysosporium BKM-F1767中漆酶的演示

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摘要

It has been widely reported that the white rot basidiomycete Phanerochaete chrysosporium, unlike most other white rot fungi, does not produce laccase, an enzyme implicated in lignin biodegradation. Our results showed that P. chrysosporium BKM-F1767 produces extracellular laccase in a defined culture medium containing cellulose (10 g/liter) and either 2.4 or 24 mM ammonium tartrate. Laccase activity was demonstrated in the concentrated extracellular culture fluids of this organism as determined by a laccase plate assay as well as a spectrophotometric assay with ABTS [2,2(prm1)-azinobis(3-ethylbenzathiazoline-6-sulfonic acid)] as the substrate. Laccase activity was observed even after addition of excess catalase to the extracellular culture fluid to destroy the endogenously produced hydrogen peroxide, indicating that the observed activity is not due to a peroxidase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by activity staining with ABTS revealed the presence of a laccase band with an estimated M(infr) of 46,500.
机译:广泛报道白腐烂担子菌Phanerochaete chrysosporium与大多数其他白腐烂真菌不同,它不产生漆酶,这是一种与木质素生物降解有关的酶。我们的结果表明,金黄色葡萄球菌BKM-F1767在含有纤维素(10克/升)和2.4或24 mM酒石酸铵的特定培养基中产生细胞外漆酶。通过漆酶平板测定和分光光度法以ABTS [2,2(prm1)-叠氮基双(3-乙基苯并噻唑啉-6-磺酸)]作为分光光度法测定,在该生物的浓缩细胞外培养液中证实了漆酶活性。基质。甚至在向细胞外培养液中添加过量的过氧化氢酶破坏内源产生的过氧化氢后,仍观察到漆酶活性,这表明观察到的活性不是由于过氧化物酶引起的。十二烷基硫酸钠-聚丙烯酰胺钠凝胶电泳,然后用ABTS进行活性染色,发现漆酶带的存在,估计M(infr)为46,500。

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