Isolation and characterization of an antifungal peptide with antiproliferative activity from seeds of Phaseolus vulgaris cv. 'Spotted Bean'

A 7.3-kDa antifungal peptide with an N-terminal sequence exhibiting remarkable homology to defensins from other leguminous plants was isolated from Phaseolus vulgaris cv. 'Spotted Bean'. The isolation procedure involved ion exchange chromatography on O-diethylaminoethyl (DEAE) cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The peptide was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and SP-Sepharose. It exerted an antifungal action on Fusarium oxysporum and Mycosphaerella arachidicola. It inhibited mycelial growth in F. oxysporum with an IC50 value of 1.8 mu M. It suppressed [methyl-H-3]-thymidine incorporation by leukemia L1210 cells and MBL2 cells with an IC50 value of 4.0 and 9.0 mu M, respectively. There was no effect on HIV-1 reverse transcriptase activity when the peptide was tested up to 0.1 mM.