title: Assessment of Schwanniomyces occidentalis as a host for protein production using the wide-range Xplor2 expression platform.
abstract: The wide-range transformation/expression platform, Xplor2, was employed for the assessment of Schwanniomyces occidentalis as a potential producer of the recombinant proteins human IFNα2a (IFNα2a) and S. occidentalis fructofuranosidase (SFfase), and its efficiency was compared to that of Arxula adeninivorans. ADE2 and URA3 genes from both yeast species were isolated, characterized and used as selection markers in combination with the IFNα2a and SFfase expression modules, which used the strong constitutive A. adeninivorans-derived TEF1 promoter. Yeast rDNA integrative expression cassettes and yeast integrative expression cassettes equipped with a selection marker and expression modules were transformed into auxotrophic S. occidentalis and A. adeninivorans strains and a quantitative comparison of the expression efficiency was made. Whilst IFNα2a was mainly accumulated extracellularly (>95 %) in A. adeninivorans, extracellular SFfase (>90 %) was detected in both yeast species. The DNA composition of the selection marker modules and expression modules, especially their open reading frame codon usage, affects auxotrophy recovery as well as protein expression. Auxotrophy recovery was only achieved with selection marker modules of the homologous gene donor yeast. The concentration of recombinant IFNα2a was fivefold higher in A. adeninivorans (1 mg L(-1)), whereas S. occidentalis accumulated 1.5- to 2-fold more SFfase (0.5 Units ml(-1)). These results demonstrate the extension of the use of the wide-range expression platform Xplor2 to another yeast species of biotechnological interest.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, Universidad Autónoma de Madrid, C/ Nicolás Cabrera 1, 28049 Madrid, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Álvaro-Benito
authors.forename: María
authors.initials: M
authors.lastname: Fernández-Lobato
authors.forename: Keith
authors.initials: K
authors.lastname: Baronian
authors.forename: Gotthard
authors.initials: G
authors.lastname: Kunze
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D010802:Phylogeny; D011995:Recombination, Genetic; D004718:Saccharomycetales / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D005656:Fungal Proteins