title: High-level production and secretion of recombinant proteins by the dimorphic yeast Arxula adeninivorans.
abstract: The non-conventional dimorphic thermo- and salt-resistant yeast Arxula adeninivorans has been developed as a host for heterologous gene expression. For assessment of the system two model genes have been selected: the GFP gene encoding the intracellular green fluorescent protein, and the HSA gene encoding the secreted human serum albumin. The expression system includes two host strains, namely A. adeninivorans LS3, which forms budding cells at 30 degrees C and mycelia at >42 degrees C, and the strain A. adeninivorans 135, which forms mycelia at temperatures as low as 30 degrees C. For expression control the constitutive A. adeninivorans-derived TEF1-promoter and S. cerevisiae-derived PHO5-terminator were selected. The basic A. adeninivorans transformation/expression vector pAL-HPH1 is further equipped with the Escherichia coli-derived hph gene conferring hygromycin B resistance and the 25S rDNA from A. adeninivorans for rDNA targeting. Transformants were obtained for both budding cells and mycelia. In both cell types similar expression levels were achieved and the GFP was localised in the cytoplasm while more than 95% of the HSA accumulated in the culture medium. In initial fermentation trials on a 200-ml shake flask scale maximal HSA product levels were observed after 96 h of cultivation.
authors.affiliation: Institut für Pflanzengenetik und Kulturpflanzenforschung, Corrensstr. 3, D-06466 Gatersleben, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wartmann
authors.forename: Erik
authors.initials: E
authors.lastname: Böer
authors.forename: Almudena Huarto
authors.initials: AH
authors.lastname: Pico
authors.forename: Heike
authors.initials: H
authors.lastname: Sieber
authors.forename: Oliver
authors.initials: O
authors.lastname: Bartelsen
authors.forename: Gerd
authors.initials: G
authors.lastname: Gellissen
authors.forename: Gotthard
authors.initials: G
authors.lastname: Kunze
mesh_terms: D015139:Blotting, Southern; D005285:Fermentation; D005819:Genetic Markers; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006921:Hygromycin B / Q000494:pharmacology*; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D004718:Saccharomycetales / Q000235:genetics* / Q000378:metabolism; D012709:Serum Albumin / Q000096:biosynthesis / Q000235:genetics; D013696:Temperature; D014170:Transformation, Genetic
chemical_list: D005819:Genetic Markers; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D012709:Serum Albumin; D049452:Green Fluorescent Proteins; D006921:Hygromycin B