The potential of Pseudozyma yeastlike epiphytes for the production of heterologous recombinant proteins

摘要: Although Basidiomycetes represent the most evolved class of fungi, they have been neglected with regard to recombinant gene expression. In this work, basidiomycetous yeasts belonging Pseudozyma spp. were studied respect their amenability heterologous protein production. Single plasmid or cotransformation experiments routinely afforded 100 200 independent transformants for two tested species Pseudozyma. Green fluorescent (GFP) was expressed in correctly folded conformation, as demonstrated by fluorescence microscopy, and hen egg white lysozyme (HEWL) its active form, revealed lytic activity on Micrococcus lysodeikticus cells. Protease analysis established that contained equivalent less extracellular protease than far ascomycetous filamentous fungi similar culture conditions. This proteolytic inhibited over 97% a combination PMSF Pepstatin A. N-glycosylation patterns native flocculosa secreted proteins comprised one few short glycan chains possess classic eukaryotic structure typical higher animal is first report Basidiomycete possesses multiple intrinsic characteristics necessary use expression system.