Institut für Biochemische Technologie und Mikrobiologie, Abteilung Mykologie, Technische Universität Wien, A-1060 Vienna, and Institut für Biochemie, Universität Wien, A-1090 Vienna, Austria.
Appl Environ Microbiol. 1988 Nov;54(11):2608-14. doi: 10.1128/aem.54.11.2608-2614.1988.
The penetration of enzymes into wood cell walls during white rot decay is an open question. A postembedding immunoelectron microscopic technique was the method of choice to answer that question. Infiltration of pine wood specimens with a concentrated culture filtrate greatly improved the labeling density and, thereby, reproducibility. Characterization of the concentrated culture filtrate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting (immunoblotting) revealed three closely spaced proteins of molecular weights about 42,000 showing immunoreactivity against anti-lignin peroxidase serum. It was shown by immunogold labeling that lignin peroxidase of Phanerochaete chrysosporium is located on the surface of the wood cell wall or within areas of heavy attack. It did not diffuse into undecayed parts of the cell wall. The reasons for preventing lignin peroxidase from penetrating wood cell walls during white rot decay are discussed.
在白腐菌腐朽过程中，酶渗透进入细胞壁是一个悬而未决的问题。包埋后免疫电镜技术是回答这个问题的首选方法。用浓缩培养液对松木标本进行渗透，极大地提高了标记密度，从而提高了重现性。浓缩培养液的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和 Western 印迹（免疫印迹）分析表明，有三种分子量约为 42000 的紧密排列的蛋白质，对木质素过氧化物酶抗血清表现出免疫反应性。免疫金标记表明，黄孢原毛平革菌的木质素过氧化物酶位于细胞壁表面或严重攻击的区域内。它没有扩散到细胞壁未腐朽的部分。讨论了木质素过氧化物酶在白腐菌腐朽过程中防止渗透进入细胞壁的原因。
