A new trypsin-chymotrypsin Inhibitor, with an N-terminal sequence showing some differences from the previously reported trypsin-chymotrypsin inhibitor, was isolated from the broad bean Vicia faba. The inhibitor was a peptide with a molecular mass of 13 kDa. It was adsorbed on Affi-gel blue gel and CM-Sepharose. It exerted antifungal activity toward Mycosphaerella arachidicola and Physalospora piricola. In addition, the trypsin-chymotrypsin inhibitor elicited a mitogenic response from mouse splenocytes and inhibited the activity of human immunodeficiency virus-1 reverse transcriptase.