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用白腐菌弗氏线虫瘤菌对小麦秸秆进行固态发酵过程中锰过氧化物酶、有机酸的产生及14C标记木质素(14C - DHP)的矿化作用

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.

作者信息

Hofrichter M, Vares T, Kalsi M, Galkin S, Scheibner K, Fritsche W, Hatakka A

机构信息

Institute of Microbiology, Friedrich Schiller University of Jena, D-07743 Jena, Germany.

出版信息

Appl Environ Microbiol. 1999 May;65(5):1864-70. doi: 10.1128/AEM.65.5.1864-1870.1999.

Abstract

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).

摘要

担子菌纲真菌弗氏线虫伞菌(Nematoloma frowardii)在小麦秸秆固态发酵(SSF)过程中产生锰过氧化物酶(MnP)作为主要的木质素分解酶。纯化后的该酶分子量为50 kDa,等电点为3.2。除MnP外,还检测到少量漆酶和木质素过氧化物酶。在固态发酵过程中,合成的14C环标记木质素(14C-DHP)被有效降解。约75%的初始放射性以14CO2形式释放,而只有6%与残留的秸秆物质相关,包括发育良好的真菌生物质。基于这一发现,我们得出结论,木质素至少可能发生了部分细胞外矿化。这一结论得到了以下事实的支持:我们在发酵秸秆中检测到高浓度的有机酸(秸秆培养物水相中的最大浓度为45 mM苹果酸、3.5 mM富马酸和10 mM草酸),这些有机酸使MnP具有活性,从而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,在天然水平的有机酸(30 mM苹果酸、5 mM富马酸)存在下,MnP实际上将部分14C-DHP转化为14CO2(占添加的初始放射性的8%)和14C标记的水溶性产物(占初始放射性的43%)。

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