title: Growth-dependent secretome of Candida utilis.
abstract: Recently, the food yeast Candida utilis has emerged as an excellent host for production of heterologous proteins. Since secretion of the recombinant product is advantageous for its purification, we characterized the secreted proteome of C. utilis. Cells were cultivated to the exponential or stationary growth phase, and the proteins in the medium were identified by MS. In parallel, a draft genome sequence of C. utilis strain DSM 2361 was determined by massively parallel sequencing. Comparisons of protein and coding sequences established that C. utilis is not a member of the CUG clade of Candida species. In total, we identified 37 proteins in the culture solution, 17 of which were exclusively present in the stationary phase, whereas three proteins were specific to the exponential growth phase. Identified proteins represented mostly carbohydrate-active enzymes associated with cell wall organization, while no proteolytic enzymes and only a few cytoplasmic proteins were detected. Remarkably, cultivation in xylose-based medium generated a protein pattern that diverged significantly from glucose-grown cells, containing the invertase Inv1 as the major extracellular protein, particularly in its highly glycosylated S-form (slow-migrating). Furthermore, cultivation without ammonium sulfate induced the secretion of the asparaginase Asp3. Comparisons of the secretome of C. utilis with those of Kluyveromyces lactis and Pichia pastoris, as well as with those of the human fungal pathogens Candida albicans and Candida glabrata, revealed a conserved set of 10 and six secretory proteins, respectively.
authors.affiliation: Molecular Mycology, Heinrich-Heine-University, Duesseldorf, Universitaetsstraße 1, 40225 Duesseldorf, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Buerth
authors.affiliation: Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.
authors.forename: Clemens J
authors.initials: CJ
authors.lastname: Heilmann
authors.affiliation: Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.
authors.forename: Frans M
authors.initials: FM
authors.lastname: Klis
authors.affiliation: Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands.
authors.forename: Chris G
authors.initials: CG
authors.lastname: de Koster
authors.affiliation: Molecular Mycology, Heinrich-Heine-University, Duesseldorf, Universitaetsstraße 1, 40225 Duesseldorf, Germany.
authors.forename: Joachim F
authors.initials: JF
authors.lastname: Ernst
authors.affiliation: Molecular Mycology, Heinrich-Heine-University, Duesseldorf, Universitaetsstraße 1, 40225 Duesseldorf, Germany.
authors.forename: Denis
authors.initials: D
authors.lastname: Tielker
mesh_terms: D002175:Candida / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D002244:Carbon / Q000378:metabolism; D003062:Codon; D005516:Food Microbiology; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D016681:Genome, Fungal; D006801:Humans; D009584:Nitrogen / Q000378:metabolism; D010802:Phylogeny; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: D003062:Codon; D005656:Fungal Proteins; D020543:Proteome; D012337:RNA, Ribosomal, 18S; D002244:Carbon; D009584:Nitrogen