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Fractionation of subcellular membranes of the secretory pathway from the peroxidase-producing white-rot fungus Phanerochaete chrysosporium
Authors:Pascal Bonnarme  Serge Moukha  Patrick Moreau  Eric Record  Laurence Lesage  Claude Cassagne  Marcel Asther
Institution:Laboratoire de Biotechnologie des Champignons Filamenteux, INRA, Facultédes Sciences de Luminy, Parc Scientifique et Technologique, 163 Avenue de Luminy, Case Postale 929, 13288 Marseille Cédex 090, France; Institut de Biochemie Cellulaire, CNRS, 1 Rue Camille Saint-Säens, 33077 Bordeaux Cedex, France
Abstract:Abstract Mycelia from the basidiomycete Phanerochaete chrysosporium , producing lignin and manganese peroxidases, were homogenized and fractionated on a sucrose gradient. The main subcellular fungal membrane fractions were successfully separated. Lipid composition analyses of the isolated membranes as well as associated marker enzymes distribution gave evidence to similarities with membranes originating from plants. Lignin and manganese peroxidases were investigated by immunodetection in subcellular fractions. Our results show that lignin and manganese peroxidases are mainly associated with Golgi apparatus vesicles and, to a lesser extent, with endoplasmic reticulum and light density vesicles, but not with plasma membranes.
Keywords:Phanerochaete chrysosporium            White-rot fungus  Secondary metabolism  Subcellular fractionation  Secretion pathway  Lignin peroxidase  Manganese peroxidase
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