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001-es BibID:BIBFORM109184
035-os BibID:(Scopus)85149121543 (WoS)000939526000001
Első szerző:Váradi Györgyi
Cím:Confirmation of the Disulfide Connectivity and Strategies for Chemical Synthesis of the Four-Disulfide-Bond-Stabilized Aspergillus giganteus Antifungal Protein, AFP / Györgyi Váradi, Gyula Batta, László Galgóczy, Dorottya Hajdu, Ádám Fizil, András Czajlik, Máté Virágh, Zoltán Kele, Vera Meyer, Sascha Jung, Florentine Marx, Gábor K. Tóth
Dátum:2023
ISSN:0163-3864 1520-6025
Megjegyzések:Emerging fungal infections require new, more efficient antifungal agents and therapies. AFP, a protein from Aspergillus giganteus with four disulfide bonds, is a promising candidate because it selectively inhibits the growth of filamentous fungi. In this work, the reduced form of AFP was prepared using native chemical ligation. The native protein was synthesized via oxidative folding with uniform protection for cysteine thiols. AFP's biological activity depends heavily on the pattern of natural disulfide bonds. Enzymatic digestion and MS analysis provide proof for interlocking disulfide topology (abcdabcd) that was previously assumed. With this knowledge, a semi-orthogonal thiol protection method was designed. By following this strategy, out of a possible 105, only 6 disulfide isomers formed and 1 of them proved to be identical with the native protein. This approach allows the synthesis of analogs for examining structure-activity relationships and, thus, preparing AFP variants with higher antifungal activity.
Tárgyszavak:Természettudományok Kémiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
folyóiratcikk
Megjelenés:Journal of Natural Products. - 86 : 4 (2023), p. 782-790. -
További szerzők:Batta Gyula (1953-) (molekula-szerkezet kutató) Galgóczy László (1950-) Hajdu Dorottya (1987-) (biológus) Fizil Ádám (1988-) (biológus) Czajlik András (1975-) (gyógyszerész) Virágh Máté Kele Zoltán Meyer, Vera Jung, Sascha Marx, Florentine Tóth Gábor K.
Pályázati támogatás:NKFIH TKP2021-EGA-32
Egyéb
NKFIH FK 134343
Egyéb
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001-es BibID:BIBFORM049587
Első szerző:Virágh Máté
Cím:Production of a defensin-like antifungal protein NFAP from Neosartorya fischeri in Pichia pastoris and its antifungal activity against filamentous fungal isolates from human infections / Máté Virágh, Dóra Vörös, Zoltán Kele, Laura Kovács, Ádám Fizil, Gergely Lakatos, Gergely Maróti, Gyula Batta, Csaba Vágvölgyi, László Galgóczy
Dátum:2014
ISSN:1046-5928
Megjegyzések:Neosartorya fischeri NRRL 181 isolate secretes a defensin-like antifungal protein (NFAP) which has a remarkable antifungal effect against ascomycetous filamentous fungi. This protein is a promising antifungal agent of biotechnological value; however in spite of the available knowledge of the nature of its 50-upstream transcriptional regulation elements, the bulk production of NFAP has not been resolved yet. In this study we carried out its heterologous expression in the yeast Pichia pastoris and investigated the growth inhibition effect exerted by the heterologous NFAP (hNFAP) on filamentous fungal isolates from human infections compared with what was caused by the native NFAP. P. pastoris KM71H transformant strain harboring the pPICZalphaA plasmid with the mature NFAP encoding gene produced the protein. The final yield of the hNFAP was sixfold compared to the NFAP produced by N. fischeri NRRL 181. Based on the signal dispersion of the amide region, it was proven that the hNFAP exists in folded state. The purified hNFAP effectively inhibited the growth of fungal isolates belonging to the Aspergillus and to the Fusarium genus, but all investigated zygomycetous strain proved to be insusceptible. There was no significant difference between the growth inhibition effect exerted by the native and the heterologous NFAP. These data indicated that P. pastoris KM71H can produce the NFAP in an antifungally active folded state. Our results provide a base for further research, e.g., investigation the connection between the protein structure and the antifungal activity using site directed mutagenesis.
Tárgyszavak:Természettudományok Biológiai tudományok idegen nyelvű folyóiratközlemény külföldi lapban
Neosartorya fischeri antifungal protein
Pichia pastoris
Antimicrobial susceptibility
Clinical fungal isolates
Ascomycetes
Zygomycetes
Megjelenés:Protein Expression and Purification 94 (2014), p. 79-84. -
További szerzők:Vörös Dóra Kele Zoltán Kovács Laura Fizil Ádám (1988-) (biológus) Lakatos Gergely Maróti Gergely Batta Gyula (1953-) (molekula-szerkezet kutató) Vágvölgyi Csaba Galgóczy László (1950-)
Pályázati támogatás:TÁMOP-4.2.4.A/2-11-1-2012-0001
TÁMOP
PD 83355
OTKA
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