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High-level expression of Aspergillus niger β-galactosidase in Ashbya gossypii
Biotechnology progress, 2014-03, Vol.30 (2), p.261-268
Magalhães, Frederico
Aguiar, Tatiana Q.
Oliveira, Carla
Domingues, Lucília
2014
Volltextzugriff (PDF)
Details
Autor(en) / Beteiligte
Magalhães, Frederico
Aguiar, Tatiana Q.
Oliveira, Carla
Domingues, Lucília
Titel
High-level expression of Aspergillus niger β-galactosidase in Ashbya gossypii
Ist Teil von
Biotechnology progress, 2014-03, Vol.30 (2), p.261-268
Ort / Verlag
Blackwell Publishing Ltd
Erscheinungsjahr
2014
Quelle
Alma/SFX Local Collection
Beschreibungen/Notizen
Ashbya gossypii has been recently considered as a host for the expression of recombinant proteins. The production levels achieved thus far were similar to those obtained with Saccharomyces cerevisiae for the same proteins. Here, the β‐galactosidase from Aspergillus niger was successfully expressed and secreted by A. gossypii from 2‐µm plasmids carrying the native signal sequence at higher levels than those secreted by S. cerevisiae laboratorial strains. Four different constitutive promoters were used to regulate the expression of β‐galactosidase: A. gossypii AgTEF and AgGPD promoters, and S. cerevisiae ScADH1 and ScPGK1 promoters. The native AgTEF promoter drove the highest expression levels of recombinant β‐galactosidase in A. gossypii, leading to 2‐ and 8‐fold higher extracellular activity than the AgGPD promoter and the heterologous promoters, respectively. In similar production conditions, the levels of active β‐galactosidase secreted by A. gossypii were up to 37 times higher than those secreted by recombinant S. cerevisiae and ∼2.5 times higher than those previously reported for the β‐galactosidase‐high producing S. cerevisiae NCYC869‐A3/pVK1.1. The substitution of glucose by glycerol in the production medium led to a 1.5‐fold increase in the secretion of active β‐galactosidase by A. gossypii. Recombinant β‐galactosidase secreted by A. gossypii was extensively glycosylated, as are the native A. niger β‐galactosidase and recombinant β‐galactosidase produced by yeast. These results highlight the potential of A. gossypii as a recombinant protein producer and open new perspectives to further optimize recombinant protein secretion in this fungus. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:261–268, 2014
Sprache
Englisch
Identifikatoren
ISSN: 8756-7938
eISSN: 1520-6033
DOI: 10.1002/btpr.1844
Titel-ID: cdi_proquest_miscellaneous_1664200206
Format
–
Schlagworte
A. gossypii GPD and TEF promoters
,
Ashbya gossypii
,
Aspergillus niger
,
Aspergillus niger β-galactosidase
,
recombinant β-galactosidase secretion
,
Saccharomyces cerevisiae
,
Saccharomyces cerevisiae PGK1 and ADH1 promoters
Weiterführende Literatur
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