title: Heterologous protein secretion by Candida utilis.
abstract: The yeast Candida utilis (also referred to as Torula) is used as a whole-cell food additive and as a recombinant host for production of intracellular molecules. Here, we report recombinant C. utilis strains secreting significant amounts of Candida antarctica lipase B (CalB). Native and heterologous secretion signals led to secretion of CalB into the growth medium; CalB was enzymatically active and it carried a short N-glycosyl chain lacking extensive mannosylation. Furthermore, CalB fusions to the C. utilis Gas1 cell wall protein led to effective surface display of enzymatically active CalB and of β-galactosidase. Secretory production in C. utilis was achieved using a novel set of expression vectors containing sat1 conferring nourseothricin resistance, which could be transformed into C. utilis, Pichia jadinii, Candida albicans, and Saccharomyces cerevisiae; C. utilis promoters including the constitutive TDH3 and the highly xylose-inducible GXS1 promoters allowed efficient gene expression. These results establish C. utilis as a promising host for the secretory production of proteins.
authors.affiliation: Department Biologie, Molekulare Mykologie, Heinrich-Heine-Universität, Universitätsstrasse 1/26.12.01, 40225 Düsseldorf, Germany.
authors.forename: Maya
authors.initials: M
authors.lastname: Kunigo
authors.forename: Christoph
authors.initials: C
authors.lastname: Buerth
authors.forename: Denis
authors.initials: D
authors.lastname: Tielker
authors.forename: Joachim F
authors.initials: JF
authors.lastname: Ernst
mesh_terms: D001709:Biotechnology / Q000379:methods; D002175:Candida / Q000235:genetics / Q000378:metabolism*; D061505:Cell Surface Display Techniques; D003470:Culture Media / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D008049:Lipase; C431349:lipase B, Candida antarctica; D001616:beta-Galactosidase