Please use this identifier to cite or link to this item: http://hdl.handle.net/1822/31895

TitleHigh-level expression of Aspergillus niger β-galactosidase in Ashbya gossypii
Author(s)Magalhães, Frederico
Aguiar, Tatiana Quinta
Oliveira, Carla Cristina Marques de
Domingues, Lucília
KeywordsAshbya gossypii
Aspergillus niger β-galactosidase
Recombinant β-galactosidase secretion
A. gossypii GPD and TEF promoters
Saccharomyces cerevisiae PGK1 and ADH1 promoters
recombinant beta-galactosidase secretion
Aspergillus niger beta-galactosidase
Issue date2014
PublisherAmerican Chemical Society
JournalBiotechnology Progress
Abstract(s)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-micron 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 approximately 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.
TypeArticle
URIhttp://hdl.handle.net/1822/31895
DOI10.1002/btpr.1844
ISSN8756-7938
e-ISSN1520-6033
Publisher versionhttp://onlinelibrary.wiley.com/journal/10.1021/(ISSN)1520-6033
Peer-Reviewedyes
AccessOpen access
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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