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 β-1,6-glucan polymer. A pure β-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 β-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 β-1,6-glucanase and was shown to be active on C. albicans cell walls allowing the release of a previously described cell wall protein.
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Boisramé, A., Gaillardin, C. Heterologous expression and characterization of a β-1,6-glucanase from Aspergillus fumigatus . Appl Microbiol Biotechnol 82, 663–669 (2009). https://doi.org/10.1007/s00253-008-1780-z
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DOI: https://doi.org/10.1007/s00253-008-1780-z