In a previous study, an antifungal protein, AFP-J, was purified from tubers of the potato ( Solanum tuberosum cv. L Jopung) and by gel filtration and HPLC. In this study, the functional peptide was characterized by partial acid digestion using HCl and HPLC. We obtained three peaks from the AFP-J, the first and third peaks were not active in the tested fungal strain. However, the second peak, which was named Potide-J, was active (MIC; 6.25 μg/mL) against Candida albicans. The amino acid sequences were analyzed by automated Edman degradation, and the amino acid sequence of Potide-J was determined to be Ala-Val-Cys-Glu-Asn-Asp-Leu-Asn-Cys-Cys. Mass spectrometry showed that its molecular mass was 1083.1 Da. Finally, we confirmed that a disulfide bond was present between Cys 3 and Cys 9 or Cys 10. Using this structure, Potide-J was synthesized via solid-phase methods. In these experiments, only the linear sequence was shown to display strong activity against Candida albicans. These results suggest that Potide-J may be an excellent candidate compound for the development of commercially applicable antibiotic agents.
Ye, X.Y.; Wang, H.X.; Ng, T.B. First chromatographic isolation of an antifungal thaumatin-like protein from French bean legumes and demonstration of its antifungal activity. Biochem. Biophys. Res. Commun 1999, 263, 130–134.
Benhamou, N.; Broglie, K.; Broglie, R.; Chet, I. Antifungal effect of bean endochitinase on Rhizoctonia solani: Ultrastructural changes and cytochemical aspects of chitin breakdown. Can. J. Microbiol 1993, 39, 318–328.
Vogelsang, R.; Barz, W. Purification, characterization and differential hormonal regulation of a β-1,3-glucanase and two chitinases from chickpea (Cicer arietinum L.). Planta 1993, 189, 60–69.
Kim, J.Y.; Park, S.C.; Kim, M.H.; Lim, H.T.; Park, Y.; Hahm, K.S. Antimicrobial activity studies on a trypsin chymotrypsin protease inhibitor obtained from potato. Biochem. Biophys. Res. Commun 2005, 330, 921–927.
Walker-Simmons, M.; Ryan, C.A. Immunological identification of proteinase inhibitors I and II in isolated tomato leaf vacuoles. Plant Physiol 1977, 60, 61–63.
Leah, R.; Tommerup, H.; Svendsen, I.; Mundy, J. Biochemical and molecular characterization of three barley seed proteins with antifungal properties. J. Biol. Chem 1991, 246, 1564–1573.
Pouvreau, L.; Gruppen, H.; van Koningsveld, G.A.; van den Broek, L.A.M.; Voragen, A.G.J. The most abundant protease inhibitor in potato tuber (cv. Elkana) is a protease inhibitor from the Kunitz family. J. Agric. Food Chem 2003, 51, 5001–5005.
Kim, J.Y.; Park, S.-C.; Hwang, I.; Cheong, H.; Nah, J.-W.; Hahm, K.-S.; Park, Y. Protease Inhibitors from Plants with Antimicrobial Activity. Int. J. Mol. Sci 2009, 10, 2860–2872.
Herzog, M.; Dorne, A.M.; Grellet, F. GASA, a gibberellin-regulated gene family from Arabidopsis thaliana related to the tomato GAST1 gene. Plant Mol. Biol 1995, 27, 743–752.
Park, C.J.; Park, C.B.; Hong, S.-S.; Lee, H.-S.; Lee, S.Y.; Kim, C. Characterization and cDNA cloning of two glycine- and histidine-rich antimicrobial peptides from the roots of shepherd’s purse, Capsella bursa-pastoris. Plant Mol. Biol 2000, 44, 187–197.
Benchekroun, A.; Alami, M.; Ghadouan, M.; Lachkar, A.; Kasmaoui, H.; Marzouk, M.; Faik, M. Urinary candidiasis revealed by ureteral obstruction: Report of 2 cases. Ann. Urol (Paris) 2000, 34, 171–174.
Jahn, B.; Martin, E.; Stueben, A.; Bhakdi, S. Susceptibility testing of Candida albicans and Aspergillus species by a simple microtiter menadione-augmented 3-(4,5-dimethyl-2-thiazolyl)- 2,5-diphenyl-2H-tetrazolium bromide assay. J. Clin. Microbiol 1995, 33, 661–667.
Pouvreau, L.; Gruppen, H.; Piersma, S.R.; van den Broek, L.A.M.; van Koningsveld, G.A.; Voragen, A.G.J. Relative abundance and inhibitory distribution of protease inhibitors in potato juice from cv. Elkana. J. Agric. Food Chem 2001, 49, 2864–2874.
