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https://hdl.handle.net/1822/31895
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Campo DC | Valor | Idioma |
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dc.contributor.author | Magalhães, Frederico | por |
dc.contributor.author | Aguiar, Tatiana Quinta | por |
dc.contributor.author | Oliveira, Carla Cristina Marques de | por |
dc.contributor.author | Domingues, Lucília | por |
dc.date.accessioned | 2014-12-12T17:29:47Z | - |
dc.date.available | 2014-12-12T17:29:47Z | - |
dc.date.issued | 2014 | - |
dc.identifier.issn | 8756-7938 | por |
dc.identifier.uri | https://hdl.handle.net/1822/31895 | - |
dc.description.abstract | 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. | por |
dc.description.sponsorship | Project AshByofactory (grant PTDC/EBB-EBI/101985/2008 - FCOMP-01-0124-FEDER-009701), MIT-Portugal Program (PhD grant SFRH/BD/39112/2007 to Tatiana Q. Aguiar) and grant SFRH/BDP/63831/2009 to Carla Oliveira | por |
dc.language.iso | eng | por |
dc.publisher | American Chemical Society | por |
dc.rights | openAccess | por |
dc.subject | Ashbya gossypii | por |
dc.subject | Aspergillus niger β-galactosidase | por |
dc.subject | Recombinant β-galactosidase secretion | por |
dc.subject | A. gossypii GPD and TEF promoters | por |
dc.subject | Saccharomyces cerevisiae PGK1 and ADH1 promoters | por |
dc.subject | recombinant beta-galactosidase secretion | por |
dc.subject | Aspergillus niger beta-galactosidase | por |
dc.title | High-level expression of Aspergillus niger β-galactosidase in Ashbya gossypii | por |
dc.type | article | - |
dc.peerreviewed | yes | por |
dc.relation.publisherversion | http://onlinelibrary.wiley.com/journal/10.1021/(ISSN)1520-6033 | por |
dc.comments | CEB17377 | por |
sdum.publicationstatus | published | por |
oaire.citationStartPage | 261 | por |
oaire.citationEndPage | 268 | por |
oaire.citationIssue | 2 | por |
oaire.citationConferencePlace | United States | - |
oaire.citationTitle | Biotechnology Progress | por |
oaire.citationVolume | 30 | por |
dc.date.updated | 2014-12-01T11:04:11Z | - |
dc.identifier.eissn | 1520-6033 | - |
dc.identifier.doi | 10.1002/btpr.1844 | por |
dc.identifier.pmid | 24851247 | por |
dc.subject.wos | Science & Technology | por |
sdum.journal | Biotechnology Progress | por |
Aparece nas coleções: | CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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document_17377_1.pdf | 392,52 kB | Adobe PDF | Ver/Abrir |