A 5443 Da
peptide with sequence homology to
defensins was purified from purple pole beans (Phaseolus vulgaris cv. 'Extra-long Purple Pole bean'). This
peptide was isolated by adsorption on an affinity chromatographic medium
Affi-Gel Blue gel and ion-exchange chromatographic media SP-
Sepharose (sulfopropyl-
Sepharose) and
Mono S and by gel filtration on Superdex
peptide. The
peptide inhibited mycelial growth in Mycosphaerella arachidicola, Helminthosporium maydis, Fusarium oxysporum, Verticillium dahliae, Rhizoctonia solani, Candida albicans and Setosphaeria turcica with an IC50 of 0.8, 0.9, 2.3, 3.2, 4.3, 4.8 and 9.8 microM respectively. Its antifungal potency was higher than that of the plant
defensin coccinin (IC50>50 microM). It induced membrane permeabilization in C. albicans as evidenced by
SYTOX Green uptake, but did not affect erythrocyte membrane permeability. It inhibited growth in M. arachidicola by inducing
chitin accumulation at hyphal
tips as was shown by
Congo Red staining. The antifungal activity was pH stable and thermostable. The
peptide inhibited the proliferation of
hepatoma (HepG2),
breast cancer (MCF7),
colon cancer (HT29) and
cervical cancer (SiHa) cells but not that of human embryonic liver (WRL68) cells. Its anti-HepG2 activity (IC50=4.1+/-0.8 microM, n=3) was higher than that of another plant
defensin, gymnin (IC50>50 microM). Its anti-MCF7 activity (IC50=8.3+/-0.3 microM, n=3) was similar to that of other plant
defensins. It reduced the activity of
HIV-1 reverse transcriptase with an IC50 of 0.5+/-0.1 microM, n=3, much more potently than other plant
defensins (IC50>40 microM). There is the possibility of using the purple pole bean
defensin for producing antifungal drugs and/or transgenic plants with fungal resistance.