A Bowman-Birk-type trypsin-chymotrypsin inhibitor from broad beans

An isolation procedure comprising affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Toyopearl, and fast protein liquid c hromatography on Mono S was used to purify a peptide from broad beans which manifested antifungal activity toward Mycosphaerella arachidicola, Fusariu m oxysporum, and Botrytis cinerea. The peptide demonstrated a molecular mas s of 7.5 kDa. N-terminal sequence analysis disclosed the identity of the an tifungal peptide to be a trypsin-chymotrypsin inhibitor. The trypsin-chymot rypsin inhibitor also exerted an inhibitory action on chymotrypsin activity and HIV-1 reverse transcriptase activity. Proliferation of murine splenocy tes was stimulated in the presence of the trypsin-chymotrypsin inhibitor. T his report constitutes the first observation of antifungal activity of a le guminous peptidic protease inhibitor. (C) 2001 Academic Press.