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Induction of manganese peroxidase and laccase by Lentinula edodes under liquid culture conditions and their isozyme detection by enzymatic staining on native-PAGE

机译:香菇在液体培养条件下对锰过氧化物酶和漆酶的诱导及其在天然PAGE上的酶法染色检测同工酶

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摘要

The white-rot basidomycete Lentinula edodes often produces the lignin-degrading enzymes manganese peroxidase (MnP; EC 1.11.1.13) and laccase (Lcc; EC 1.10.3.2) in sawdust-based media. In the present study, MnP from L. edodes was induced under liquid culture supplemented with sawdust extracts of Castanopsis cuspidata. Lcc activity was induced by the addition of 2 mM CuSOp"5HO into the same media 7 days after initial inoculation. Phenoloxidase enzymes were distinguished by polyacrylamide gel electrophoresis (native-PAGE), followed by sequential enzymatic staining with an improved staining solution. The isozyme bands detected under MnP-induced conditions were identified as manganese peroxidase (lemnp2) and bands detected under Lcc-induced conditions were identified as laccase (lcc1) by Q-TOF mass spectrometry.
机译:白腐担子菌香菇通常在基于锯末的培养基中产生木质素降解酶锰过氧化物酶(MnP; EC 1.11.1.13)和漆酶(Lcc; EC 1.10.3.2)。在本研究中,在液体培养中补充了来自锥栗的木屑提取物,从而诱导了香菇的MnP。初次接种7天后,在同一培养基中添加2 mM CuSOp“ 5HO,从而诱导Lcc活性。通过聚丙烯酰胺凝胶电泳(天然PAGE)区分酚氧化酶,然后用改良的染色液进行顺序酶促染色。通过Q-TOF质谱将在MnP诱导条件下检测到的谱带鉴定为锰过氧化物酶(lemnp2),在Lcc诱导条件下检测到的谱带鉴定为漆酶(lcc1)。

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