title: Heterologous expression and characterization of a beta-1,6-glucanase from Aspergillus fumigatus.
abstract: The cell wall of Candida albicans is composed of mannoproteins associated to glycan polymers. Most of these proteins are retained in this compartment through a phosphodiester linkage between a remnant of their glycosylphosphatidylinositol anchor and the beta-1,6-glucan polymer. A pure beta-1,6-glucanase is thus required in order to release them. In this paper, we report the expression/secretion by the yeast Yarrowia lipolytica of an Aspergillus fumigatus enzyme homologous to previously described beta-1,6-glucanases. The coding sequence was expressed under the control of a strong promoter and the recombinant enzyme was targeted to the secretory pathway using the signal sequence of a well-known major secretory protein in this host. Addition of a FLAG epitope at the C-terminus allowed its efficient purification from culture supernatant following batch adsorption. The purified enzyme was characterized as a beta-1,6-glucanase and was shown to be active on C. albicans cell walls allowing the release of a previously described cell wall protein.
authors.affiliation: Laboratoire Microbiologie et Génétique Moléculaire, UMR AgroParisTech, INRA, CNRS, CBAI, 78850 Thiverval-Grignon, France. boisrame@grignon.inra.fr
authors.forename: A
authors.initials: A
authors.lastname: Boisramé
authors.forename: C
authors.initials: C
authors.lastname: Gaillardin
mesh_terms: D001232:Aspergillus fumigatus / Q000201:enzymology*; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005936:Glucans / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008970:Molecular Weight; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D025062:Yarrowia / Q000235:genetics* / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D005936:Glucans; D011993:Recombinant Fusion Proteins; D006026:Glycoside Hydrolases; C020105:endo-1,6-beta-glucanase