, 2006). Recently, fungicidal as well as bactericidal AMPs have been found and isolated from a wide range of organisms, including amphibians, invertebrates, plants, insects and mammals (Hwang & Vogel, 1998; Zasloff, 2002). Papiliocin (RWKIFKKIEKVGRNVRDGIIKAGPAVAVVGQAATVVK-NH2) is a 37-residue peptide isolated from the larvae of the swallowtail butterfly Papilio xuthus (Kim et al., 2010). [Correction added on 24 August after online publication:
38 corrected to 37 in this sentence and also in the Abstract; also a G has been removed from the end of the amino acid sequence.] In this study, the antifungal activity and mechanism of papiliocin were investigated and its antifungal properties were suggested. Peptide synthesis was carried out by Anygen Co. (Gwangju, Korea). The following procedures for peptide synthesis are offered by Anygen Co. Dabrafenib research buy The assembly of peptides consisted of a 60-min cycle for each residue at ambient temperature as follows: (1) the 2-chlorotrityl (or 4-methylbenzhydrylamine amide) resin was charged to a reactor and then washed with dichloromethane and N,N-dimethylformamide (DMF), respectively, and (2) a coupling
step with vigorous shaking using a 0.14 mM solution of Fmoc-l-amino acids and Fmoc-l-amino acids preactivated for approximately 60 min with a 0.1 mM solution of 0.5 M HOBt/DIC in DMF. Finally, the peptide was cleaved from the resin using a trifluoroacetic acid (TFA) cocktail solution at ambient Epacadostat nmr temperature (Merrifield, 1986; Sheppard, 2003). Analytical and preparative reverse-phase HPLC runs were performed using a Shimadzu 20A or 6A gradient system. Data were collected using an SPD-20A detector at 230 nm. Chromatographic separations were achieved with a 1%/min linear gradient of
buffer B in A (A=0.1% TFA in H2O; B=0.1% TFA in CH3CN) Histidine ammonia-lyase over 40 min at flow rates of 1 and 8 mL min−1 using Shimadzu C18 analytical (5 μm, 0.46 cm × 25 cm) and preparative C18 (10 μm, 2.5 cm × 25 cm) columns, respectively. Aspergillus flavus (KCTC 1375), Aspergillus fumigatus (KCTC 6145), Aspergillus parasiticus (KCTC 6598), Malassezia furfur (KCTC 7744), Trichophyton rubrum (KCTC 6345) and Trichosporon beigelii (KCTC 7707) were obtained from the Korean Collection for Type Cultures (KCTC) (Daejeon, Korea). Candida albicans (TIMM 1768) was obtained from the Center for Academic Societies (Osaka, Japan). Candida albicans (ATCC 90028) and Candida parapsilosis (ATCC 22019) were obtained from the American Type Culture Collection (ATCC) (Manassas, VA). Fungal cells were cultured in YPD broth (Difco), containing yeast extract, peptone and dextrose (50 g L−1), with aeration at 28 °C.