In situ localization of manganese peroxidase production in mycelial pellets of Phanerochaete chrysosporium

1 Laboratoire de Physiologie et Ecologie Microbienne, Faculté des Sciences, Université Libre de Bruxelles, c/o Institut Pasteur, 642 Rue Engeland, B-1180 Brussels, Belgium 2 Laboratory of Actinomycetes and Fungi imperfecti, National Institute of Hygiene, Warsaw, Poland 3 Department of Microbiology and Biotechnology, Faculty of Sciences, University of Debrecen, Debrecen, Hungary Correspondence M. J. Penninckx upemulb{at}resulb.ulb.ac.be The ultrastructure of Phanerochaete chrysosporium hyphae from pellets in submerged liquid cultures was investigated in order to learn more about the interrelation between fungal architecture and manganese peroxidase (MnP) production. At day 2 of cultivation, some subapical regions of hyphae in the outer and middle zones of the pellet initiated differentiation into intercalary thick-walled chlamydospore-like cells of about 10 µm diameter. At the periphery of the cytoplasm of these cells, a large number of mitochondria and Golgi-like vesicles were observed. The sites of MnP production were localized at different stages of cultivation by an immunolabelling procedure. The immunomarker of MnP was mainly concentrated in the chlamydospore-like cells and principally distributed in Golgi-like vesicles located at the periphery of the cytoplasm. The apices of hyphae in the outer layer of the pellets were apparently minor sites of MnP production. Maximal MnP release into the culture supernatant coincided with apparent autolysis of the chlamydospore-like cells. Production of extracellular autolytic chitinase and protease coincided with the disappearance of these structures from the pellets. The chlamydospore-like cells observed in the mycelial pellets of P. chrysosporium could be metabolically active entities operating as an enzyme reservoir, delivering their content into the surrounding medium possibly by an enzyme-mediated autolytic process. Abbreviations: LiP, lignin peroxidase; MnP, manganese peroxidase