caa a22 4500 605498830 CHVBK 20210128100548.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6541-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6541-1 Concatemerization increases the inhibitory activity of short, cell-penetrating, cationic and tryptophan-rich antifungal peptides [Elektronische Daten] [Belén López-García, Eleonora Harries, Lourdes Carmona, Lidia Campos-Soriano, José López, Paloma Manzanares, Mónica Gandía, María Coca, Jose Marcos] There are short cationic and tryptophan-rich antifungal peptides such as the hexapeptide PAF26 (RKKWFW) that have selective toxicity and cell penetration properties against fungal cells. This study demonstrates that concatemeric peptides with tandem repeats of the heptapeptide PAF54 (which is an elongated PAF26 sequence) show increased fungistatic and bacteriostatic activities while maintaining the absence of hemolytic activity of the monomer. The increase in antimicrobial activity of the double-repeated PAF sequences (diPAFs), compared to the nonrepeated PAF, was higher (4-8-fold) than that seen for the triple-repeated sequences (triPAFs) versus the diPAFs (2-fold). However, concatemerization diminished the fungicidal activity against quiescent spores of the filamentous fungus Penicillium digitatum. Peptide solubility and sensitivity to proteolytic degradation were affected by the design of the concatemers: incorporation of the AGPA sequence hinge to separate PAF54 repeats increased solubility while the C-terminal addition of the KDEL sequence decreased in vitro stability. These results led to the design of the triPAF sequence PAF102 of 30 amino acid residues, with increased antimicrobial activity and minimal inhibitory concentration (MIC) value of 1-5μM depending on the fungus. Further characterization of the mode-of-action of PAF102 demonstrated that it colocalizes first with the fungal cell wall, it is thereafter internalized in an energy dependent manner into hyphal cells of the filamentous fungus Fusarium proliferatum, and finally kills hyphal cells intracellularly. Therefore, PAF102 showed mechanistic properties against fungi similar to the parental PAF26. These observations are of high interest in the future development of PAF-based antimicrobial molecules optimized for their production in biofactories. Springer-Verlag Berlin Heidelberg, 2015 Antimicrobial nationallicence Antifungal peptide nationallicence Cell-penetrating peptide nationallicence Phytopathogen nationallicence Fungal cell wall nationallicence López-García Belén Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus UAB, 08193, Bellaterra, Barcelona, Spain aut Harries Eleonora Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut Carmona Lourdes Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut Campos-Soriano Lidia Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus UAB, 08193, Bellaterra, Barcelona, Spain aut López José Biotechnology Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut Manzanares Paloma Biotechnology Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut Gandía Mónica Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut Coca María Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus UAB, 08193, Bellaterra, Barcelona, Spain aut Marcos Jose Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 8011-8021 0175-7598 99:19<8011 2015 99 253 https://doi.org/10.1007/s00253-015-6541-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6541-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- López-García Belén Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus UAB, 08193, Bellaterra, Barcelona, Spain aut NATIONALLICENCE 700 1- Harries Eleonora Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut NATIONALLICENCE 700 1- Carmona Lourdes Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut NATIONALLICENCE 700 1- Campos-Soriano Lidia Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus UAB, 08193, Bellaterra, Barcelona, Spain aut NATIONALLICENCE 700 1- López José Biotechnology Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut NATIONALLICENCE 700 1- Manzanares Paloma Biotechnology Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut NATIONALLICENCE 700 1- Gandía Mónica Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut NATIONALLICENCE 700 1- Coca María Centre for Research in Agricultural Genomics (CRAG), CSIC-IRTA-UAB-UB. Edifici CRAG, Campus UAB, 08193, Bellaterra, Barcelona, Spain aut NATIONALLICENCE 700 1- Marcos Jose Food Science Department, Instituto de Agroquímica y Tecnología de Alimentos (IATA) - CSIC, Avda. Agustín Escardino 7, 46980, Paterna, Valencia, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 8011-8021 0175-7598 99:19<8011 2015 99 253