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Assessment of Schwanniomyces occidentalis as a host for protein production using the wide-range Xplor®2 expression platform
Ist Teil von
Applied microbiology and biotechnology, 2013-05, Vol.97 (10), p.4443-4456
Ort / Verlag
Berlin/Heidelberg: Springer-Verlag
Erscheinungsjahr
2013
Quelle
Alma/SFX Local Collection
Beschreibungen/Notizen
The wide-range transformation/expression platform, Xplor®2, 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
.
ADE
2 and
URA
3 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 Xplor®2 to another yeast species of biotechnological interest.