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Production of Manganese Peroxidase and Organic Acids and Mineralization of 14C-Labelled Lignin (14C-DHP) during Solid-State Fermentation of Wheat Straw with the White Rot Fungus Nematoloma frowardii

机译:白腐真菌线虫线虫固态发酵小麦秸秆过程中锰过氧化物酶和有机酸的产生及14C-Labelled木质素(14C-DHP)的矿化

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摘要

The basidiomycetous fungus Nematoloma frowardii produced manganese peroxidase (MnP) as the predominant ligninolytic enzyme during solid-state fermentation (SSF) of wheat straw. The purified enzyme had a molecular mass of 50 kDa and an isoelectric point of 3.2. In addition to MnP, low levels of laccase and lignin peroxidase were detected. Synthetic 14C-ring-labelled lignin (14C-DHP) was efficiently degraded during SSF. Approximately 75% of the initial radioactivity was released as 14CO2, while only 6% was associated with the residual straw material, including the well-developed fungal biomass. On the basis of this finding we concluded that at least partial extracellular mineralization of lignin may have occurred. This conclusion was supported by the fact that we detected high levels of organic acids in the fermented straw (the maximum concentrations in the water phases of the straw cultures were 45 mM malate, 3.5 mM fumarate, and 10 mM oxalate), which rendered MnP effective and therefore made partial direct mineralization of lignin possible. Experiments performed in a cell-free system, which simulated the conditions in the straw cultures, revealed that MnP in fact converted part of the 14C-DHP to 14CO2 (which accounted for up to 8% of the initial radioactivity added) and 14C-labelled water-soluble products (which accounted for 43% of the initial radioactivity) in the presence of natural levels of organic acids (30 mM malate, 5 mM fumarate).
机译:在小麦秸秆的固态发酵(SSF)中,担子菌真菌frowardii产生了锰过氧化物酶(MnP)作为主要的木质素分解酶。纯化的酶的分子量为50kDa,等电点为3.2。除了MnP,还检测到低水平的漆酶和木质素过氧化物酶。合成的 14 C环标记的木质素( 14 C-DHP)在SSF中被有效降解。大约75%的初始放射性以 14 CO2的形式释放出来,而只有6%与残留的稻草材料有关,包括发达的真菌生物质。根据这一发现,我们得出结论,木质素可能至少发生了部分细胞外矿化。我们在发酵的秸秆中检测到高水平的有机酸(秸秆培养物的水相中最大浓度为苹果酸45 mM,富马酸盐3.5 mM和草酸盐10 mM)这一事实支持了这一点,从而使MnP有效因此使木质素的部分直接矿化成为可能。在模拟秸秆培养条件的无细胞系统中进行的实验表明,MnP实际上将 14 C-DHP的一部分转化为 14 CO2(在天然水平的有机酸存在下(30)最多占最初添加的放射性的8%)和 14 C标记的水溶性产品(占最初添加的放射性的43%)苹果酸mM,富马酸5 mM)。

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