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利用透射电子显微镜和免疫金标记技术研究杂色云芝降解实木板和木屑过程中木质素过氧化物酶的胞内外定位。

Intra- and Extracellular Localization of Lignin Peroxidase during the Degradation of Solid Wood and Wood Fragments by Phanerochaete chrysosporium by Using Transmission Electron Microscopy and Immuno-Gold Labeling.

机构信息

Department of Forest Products, Swedish University of Agricultural Sciences, Box 7008, S-750 07 Uppsala, and STFI, S-114 86 Stockholm, Sweden.

出版信息

Appl Environ Microbiol. 1989 Apr;55(4):871-81. doi: 10.1128/aem.55.4.871-881.1989.

Abstract

The distribution of lignin peroxidase during degradation of both wood and woody fragments by the white rot fungus Phanerochaete chrysosporium was investigated by using anti-lignin peroxidase in conjunction with postembedding transmission electron microscopy and immuno-gold labeling techniques. The enzyme was localized in the peripheral regions of the fungal cell cytoplasm in association with the cell membrane, fungal cell wall, and extracellular slime materials. In solid wood, lignin peroxidase was detected in low concentrations associated with both superficial and degradation zones within secondary cell walls undergoing fungal attack. A similar but much greater level of extracellular peroxidase activity was associated with wood fragments degraded by the fungus grown under liquid culture conditions optimal for production of the enzyme. Efforts to infiltrate degraded wood pieces with high levels of lignin peroxidase showed the enzyme to be restricted to superficial regions of wood decay and to penetrate wood cell walls only where the wall structure had been modified. In this respect the enzyme was able to penetrate characteristic zones of degradation within the secondary walls of fibers to sites of lignin attack. This suggests a possibility for a close substrate-enzyme association during wood cell wall degradation.

摘要

用抗木质素过氧化物酶结合包埋后透射电子显微镜和免疫胶体金标记技术,研究了白腐菌糙皮侧耳降解木材和木质碎片过程中木质素过氧化物酶的分布。该酶定位于真菌细胞质的外周区域,与细胞膜、真菌细胞壁和细胞外粘液物质有关。在固体木材中,木质素过氧化物酶与受真菌攻击的次生细胞壁的表层和降解区结合,浓度较低。在液体培养条件下,真菌生长最佳,有利于酶的产生,与木质碎片降解相关的细胞外过氧化物酶活性水平相似,但要高得多。渗透到具有高浓度木质素过氧化物酶的降解木材块的努力表明,该酶仅限于木材腐烂的表面区域,并且仅在细胞壁结构已经改变的地方穿透木材细胞壁。在这方面,该酶能够穿透纤维次生壁的特征降解区域到达木质素攻击部位。这表明在细胞壁降解过程中存在紧密的底物-酶结合的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/684d/184217/5bf01804ab6d/aem00097-0112-a.jpg

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