Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum

Abstract Filamentous fungi encode distinct antifungal proteins (AFPs) that offer great potential to develop new antifungals. Fungi are considered immune to their own AFPs as occurs in Penicillium chrysogenum, the producer of the well-known PAF. The Penicillium digitatum genome encodes only one afp g...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sandra Garrigues, Mónica Gandía, Crina Popa, Attila Borics, Florentine Marx, María Coca, Jose F. Marcos, Paloma Manzanares
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/15db684c777c4b7696d438354dfbf313
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:15db684c777c4b7696d438354dfbf313
record_format	dspace
spelling	oai:doaj.org-article:15db684c777c4b7696d438354dfbf3132021-12-02T15:05:52ZEfficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum10.1038/s41598-017-15277-w2045-2322https://doaj.org/article/15db684c777c4b7696d438354dfbf3132017-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-15277-whttps://doaj.org/toc/2045-2322Abstract Filamentous fungi encode distinct antifungal proteins (AFPs) that offer great potential to develop new antifungals. Fungi are considered immune to their own AFPs as occurs in Penicillium chrysogenum, the producer of the well-known PAF. The Penicillium digitatum genome encodes only one afp gene (afpB), and the corresponding protein (AfpB) belongs to the class B phylogenetic cluster. Previous attempts to detect AfpB were not successful. In this work, immunodetection confirmed the absence of AfpB accumulation in wild type and previous recombinant constitutive P. digitatum strains. Biotechnological production and secretion of AfpB were achieved in P. digitatum with the use of a P. chrysogenum-based expression cassette and in the yeast Pichia pastoris with the α-factor signal peptide. Both strategies allowed proper protein folding, efficient production and single-step purification of AfpB from culture supernatants. AfpB showed antifungal activity higher than the P. chrysogenum PAF against the majority of the fungi tested, especially against Penicillium species and including P. digitatum, which was highly sensitive to the self-AfpB. Spectroscopic data suggest that native folding is not required for activity. AfpB also showed notable ability to withstand protease and thermal degradation and no haemolytic activity, making AfpB a promising candidate for the control of pathogenic fungi.Sandra GarriguesMónica GandíaCrina PopaAttila BoricsFlorentine MarxMaría CocaJose F. MarcosPaloma ManzanaresNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sandra Garrigues Mónica Gandía Crina Popa Attila Borics Florentine Marx María Coca Jose F. Marcos Paloma Manzanares Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
description	Abstract Filamentous fungi encode distinct antifungal proteins (AFPs) that offer great potential to develop new antifungals. Fungi are considered immune to their own AFPs as occurs in Penicillium chrysogenum, the producer of the well-known PAF. The Penicillium digitatum genome encodes only one afp gene (afpB), and the corresponding protein (AfpB) belongs to the class B phylogenetic cluster. Previous attempts to detect AfpB were not successful. In this work, immunodetection confirmed the absence of AfpB accumulation in wild type and previous recombinant constitutive P. digitatum strains. Biotechnological production and secretion of AfpB were achieved in P. digitatum with the use of a P. chrysogenum-based expression cassette and in the yeast Pichia pastoris with the α-factor signal peptide. Both strategies allowed proper protein folding, efficient production and single-step purification of AfpB from culture supernatants. AfpB showed antifungal activity higher than the P. chrysogenum PAF against the majority of the fungi tested, especially against Penicillium species and including P. digitatum, which was highly sensitive to the self-AfpB. Spectroscopic data suggest that native folding is not required for activity. AfpB also showed notable ability to withstand protease and thermal degradation and no haemolytic activity, making AfpB a promising candidate for the control of pathogenic fungi.
format	article
author	Sandra Garrigues Mónica Gandía Crina Popa Attila Borics Florentine Marx María Coca Jose F. Marcos Paloma Manzanares
author_facet	Sandra Garrigues Mónica Gandía Crina Popa Attila Borics Florentine Marx María Coca Jose F. Marcos Paloma Manzanares
author_sort	Sandra Garrigues
title	Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
title_short	Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
title_full	Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
title_fullStr	Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
title_full_unstemmed	Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
title_sort	efficient production and characterization of the novel and highly active antifungal protein afpb from penicillium digitatum
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/15db684c777c4b7696d438354dfbf313
work_keys_str_mv	AT sandragarrigues efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT monicagandia efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT crinapopa efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT attilaborics efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT florentinemarx efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT mariacoca efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT josefmarcos efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum AT palomamanzanares efficientproductionandcharacterizationofthenovelandhighlyactiveantifungalproteinafpbfrompenicilliumdigitatum
_version_	1718388627792199680

Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum

Ejemplares similares