Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum
Garrigues, Sandra (Instituto de Agroquímica y Tecnología de Alimentos)
Gandía, Mónica (Instituto de Agroquímica y Tecnología de Alimentos)
Popa, Crina (Centre de Recerca en Agrigenòmica)
Borics, Attila (Magyar Tudományos Akadémia. Szegedi Biológiai Központ)
Marx, Florentine (Medizinische Universität Innsbruck)
Coca López, María (Centre de Recerca en Agrigenòmica)
Marcos, Jose F. (Instituto de Agroquímica y Tecnología de Alimentos)
Manzanares, Paloma (Instituto de Agroquímica y Tecnología de Alimentos)

Date:	2017
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.
Grants:	Ministerio de Economía y Competitividad BIO2015-68790-C2-1-R Ministerio de Economía y Competitividad BIO2015-68790-C2-2-R Ministerio de Educación, Cultura y Deporte FPU13/04584
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Antifungal agents ; Biotechnology
Published in:	Scientific reports, Vol. 7 (2017) , art. 14663, ISSN 2045-2322

DOI: 10.1038/s41598-017-15277-w
PMID: 29116156

13 p, 3.5 MB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Experimental sciences > CRAG (Centre for Research in Agricultural Genomics)
Articles > Research articles
Articles > Published articles