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https://hdl.handle.net/1822/31895
Título: | High-level expression of Aspergillus niger β-galactosidase in Ashbya gossypii |
Autor(es): | Magalhães, Frederico Aguiar, Tatiana Quinta Oliveira, Carla Cristina Marques de Domingues, Lucília |
Palavras-chave: | Ashbya 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 |
Data: | 2014 |
Editora: | American Chemical Society |
Revista: | Biotechnology Progress |
Resumo(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. |
Tipo: | Artigo |
URI: | https://hdl.handle.net/1822/31895 |
DOI: | 10.1002/btpr.1844 |
ISSN: | 8756-7938 |
e-ISSN: | 1520-6033 |
Versão da editora: | http://onlinelibrary.wiley.com/journal/10.1021/(ISSN)1520-6033 |
Arbitragem científica: | yes |
Acesso: | Acesso aberto |
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 |