The radish defensins RsAFP1 and RsAFP2 act synergistically with caspofungin against Candida albicans biofilms |
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| Affiliation: | 1. Centre of Microbial and Plant Genetics, KU Leuven, Kasteelpark Arenberg 20, 3001 Heverlee, Belgium;2. Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia;3. Department of Materials Engineering, KU Leuven, Kasteelpark Arenberg 44, 3001 Heverlee, Belgium;4. Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Ghent, Belgium;1. Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, Institute of Health Biotechnology, International Collaborative Research Center for Health Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People''s Republic of China;2. Obesita & Algaegen LLC, College Station, TX 77845, USA;3. Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science & Technology), Ministry of Education, Tianjin Key Lab of Industrial Microbiology, Tianjin University of Science and Technology, Tianjin, 300457, People''s Republic of China;4. Tianjin Haifa Sea Food Industrial Development Co., LTD, People''s Republic of China;1. Unidad de Bioquímica y Biología Molecular de Plantas. Centro de Investigación Científica de Yucatán A.C., Calle 43 No. 130, Col. Chuburná de Hidalgo, Mérida, Yucatán 97205, México;2. Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, UNAM. Apartado Postal 510-3, Cuernavaca, Morelos, 61500, México;1. Department of Microbiology, Faculty of Pharmacy, University of Santiago de Compostela, Spain;2. Faculty of Veterinary Science and Bosch Institute, University of Sydney, NSW 2006, Australia;3. Department of Food Science and Technology, Faculty of Veterinary, University of Santiago de Compostela, Spain;1. Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, 117997 Moscow, Russia;2. Moscow Institute of Physics and Technology (State University), Department of Physicochemical Biology and Biotechnology, Institutskii per., 9, 141700 Dolgoprudny, Moscow Region, Russia |
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| Abstract: | The radish defensin RsAFP2 was previously characterized as a peptide with potent antifungal activity against several plant pathogenic fungi and human pathogens, including Candida albicans. RsAFP2 induces apoptosis and impairs the yeast-to-hypha transition in C. albicans. As the yeast-to-hypha transition is considered important for progression to mature biofilms, we analyzed the potential antibiofilm activity of recombinant (r)RsAFP2, heterologously expressed in Pichia pastoris, against C. albicans biofilms. We found that rRsAFP2 prevents C. albicans biofilm formation with a BIC-2 (i.e., the minimal rRsAFP2 concentration that inhibits biofilm formation by 50% as compared to control treatment) of 1.65 ± 0.40 mg/mL. Moreover, biofilm-specific synergistic effects were observed between rRsAFP2 doses as low as 2.5 μg/mL to 10 μg/mL and the antimycotics caspofungin and amphotericin B, pointing to the potential of RsAFP2 as a novel antibiofilm compound. In addition, we characterized the solution structure of rRsAFP2 and compared it to that of RsAFP1, another defensin present in radish seeds. These peptides have similar amino acid sequences, except for two amino acids, but rRsAFP2 is more potent than RsAFP1 against planktonic and biofilm cultures. Interestingly, as in case of rRsAFP2, also RsAFP1 acts synergistically with caspofungin against C. albicans biofilms in a comparable low dose range as rRsAFP2. A structural comparison of both defensins via NMR analysis revealed that also rRsAFP2 adopts the typical cysteine-stabilized αβ-motif of plant defensins, however, no structural differences were found between these peptides that might result in their differential antifungal/antibiofilm potency. This further suggests that the conserved structure of RsAFP1 and rRsAFP2 bears the potential to synergize with antimycotics against C. albicans biofilms. |
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| Keywords: | plant defensin RsAFP1 RsAFP2 recombinant protein production NMR analysis fungal biofilm |
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