8452e2ef87845d327aa17bd4baa6dc6b pmc_29116156.pdf 550560d4e9bc16f1fac8879ba215807dc62690b3 pmc_29116156.pdf 4b825f2bbea267ef65fdb3f13c5451ede03ab4689d5929aa24321bf393358f95 pmc_29116156.pdf Title: Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum Subject: Scientific Reports, doi:10.1038/s41598-017-15277-w Keywords: Author: Sandra Garrigues Creator: Springer Producer: iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) CreationDate: Tue Oct 31 10:17:51 2017 CET ModDate: Wed Nov 1 12:40:27 2017 CET Tagged: no UserProperties: no Suspects: no Form: AcroForm JavaScript: no Pages: 13 Encrypted: no Page size: 595.276 x 782.362 pts Page rot: 0 File size: 3657054 bytes Optimized: no PDF version: 1.4 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ERQIXF+Corbel CID TrueType Identity-H yes yes yes 588 0 XYPADT+TimesNewRomanPSMT Type 1C WinAnsi yes yes yes 589 0 MinionPro-Regular Type 1C Custom yes no yes 74 0 GCYHIN+Corbel-BoldItalic Type 1C WinAnsi yes yes yes 68 0 MinionPro-It Type 1C Custom yes no yes 67 0 GJXBXS+Corbel2 Type 1C WinAnsi yes yes yes 71 0 AGOFTN+Corbel-Bold CID TrueType Identity-H yes yes yes 73 0 EuclidSymbol-Bold Type 1C Custom yes no yes 590 0 WWPCPQ+Corbel-Bold2 Type 1C WinAnsi yes yes yes 591 0 WWPCPQ+Corbel-Bold2 Type 1C WinAnsi yes yes yes 517 0 GJXBXS+Corbel2 Type 1C WinAnsi yes yes yes 518 0 ERQIXF+Corbel CID TrueType Identity-H yes yes yes 632 0 MyriadPro-It Type 1C WinAnsi yes no yes 519 0 STIXGeneral-Italic Type 1C Custom yes no yes 66 0 MinionPro-Bold Type 1C WinAnsi yes no yes 69 0 EuclidSymbol Type 1C Custom yes no yes 70 0 SANXLD+STIXGeneral-Regular Type 1C Custom yes yes yes 72 0 GJXBXS+Corbel2 Type 1C WinAnsi yes yes yes 495 0 ERQIXF+Corbel CID TrueType Identity-H yes yes yes 508 0 WWPCPQ+Corbel-Bold2 Type 1C WinAnsi yes yes yes 525 0 GJXBXS+Corbel2 Type 1C WinAnsi yes yes yes 522 0 ERQIXF+Corbel CID TrueType Identity-H yes yes yes 524 0 STIXGeneral-Bold Type 1C Custom yes no yes 145 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-02-08 02:44:47 CET RepresentationInformation: pmc_29116156.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2020-08-10 03:39:41 CEST Size: 3657054 Format: PDF Version: 1.4 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 702 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: false CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseOutlines Language: EN Outlines: Item: Title: Efficient production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum ... Children: Item: Title: Results Children: Item: Title: Recombinant production of AfpB in P. digitatum and P. pastoris. Item: Title: Single-step cationic exchange chromatography allowed AfpB purification from P. digitatum and P. pastoris culture supernatan ... Item: Title: Immunodetection confirmed the absence of AfpB in the parental strain. Item: Title: Mass spectrometry revealed different processing of PAF and AfpB SP-pro sequence. Item: Title: Electronic circular dichroism spectroscopy assays revealed incomplete refolding capability of the AfpB* variant. Item: Title: AfpB was highly active against Penicillium species including P. digitatum. Item: Title: AfpB showed high protease and thermal resistance. Item: Title: AfpB showed no haemolytic activity against human red blood cells. Item: Title: Discussion Item: Title: Methods Children: Item: Title: Strains, media and culture conditions. Item: Title: Generation of the P. digitatum AfpB producer strains. Item: Title: Generation of the P. pastoris AfpB producer strains. Item: Title: Protein production and purification. Item: Title: Western blot. Item: Title: MALDI-TOF-MS. Item: Title: ECD spectroscopy. Item: Title: Protein stability assays. Item: Title: Antimicrobial activity assays. Item: Title: Haemolytic assays. Item: Title: Fruit infection assays. Item: Title: Data availability. Item: Title: Acknowledgements Item: Title: Figure 1 Schematic representation of the expression systems used to obtain AfpB producer strains. Item: Title: Figure 2 Growth of P. Item: Title: Figure 3 Analyses of supernatants of AfpB-producing strains and purified proteins by SDS-PAGE and Western blot. Item: Title: Figure 4 MALDI-TOF MS analyses. Item: Title: Figure 5 ECD spectra of the two recombinant protein variants produced in P. Item: Title: Figure 6 In vitro inhibitory activity of AfpB and PAF against filamentous fungi. Item: Title: Figure 7 Effect of heat treatment on the antifungal activity of PAF and AfpB. Item: Title: Figure 8 Haemolytic activity of the two AfpB variants and PAF. 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production and characterization of the novel and highly active antifungal protein AfpB from Penicillium digitatum Sandra Garrigues Mamp#x000F3;nica Gandamp#x000ED;a Crina Popa Attila Borics Florentine Marx Maramp#x000ED;a Coca Jose F. Marcos Paloma Manzanares 10.1038/s41598-017-15277-w http://dx.doi.org/10.1038/s41598-017-15277-w journal Scientific Reports © 0, The Author(s) 2045-2322 10.1038/s41598-017-15277-w Springer 2017-11-01T12:40:27+01:00 2017-10-31T14:47:51+05:30 2017-11-01T12:40:27+01:00 True iText® 5.3.5 ©2000-2012 1T3XT BVBA (AGPL-version) VoR uuid:db5bda69-6ac4-4d52-bd26-907e09b7fd99 uuid:6f834874-e4a4-427f-9035-2d9a0f817a63 default 1 converted uuid:e68502fc-a0ad-45df-8bcf-36e16fbe65b5 converted to PDF/A-2b pdfToolbox 2017-10-31T14:48:32+05:30 2 B http://ns.adobe.com/pdf/1.3/ pdf Adobe PDF Schema internal A name object indicating whether the document has been modified to include trapping information Trapped Text http://ns.adobe.com/xap/1.0/mm/ xmpMM XMP Media Management Schema internal UUID based identifier for specific incarnation of a document InstanceID URI internal The common identifier for all versions and renditions of a document. 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