The subtilisin inhibitor (SI), isolated from black beans (Phaseolus vagaris) has a molecular weight of 8918 and it contains four half cystine residues. The N‐terminal residue is arginine and the C‐terminal is lysine. The subtilisin inhibiting site is composed of an Ala‐Leu or Ile bond. This site interacts in addition to subtilisin (S) with elastase from human leukocytes (HLE), both of which compete for the inhibitor. Also, HLE‐cleaved SI (SIHLE) is inactive against HLE as well as against S. It was shown that the inhibition of S by SI resembles a temporary inhibition, involving a very slow release of enzyme activity. The trypsin reactive site is a Lys‐Val bond, and its interaction with bovine trypsin is very weak. No complexes of SI or trypsin‐cleaved SI (SIT) with trypsin could be detected on cellulose‐acetate membrane electrophoresis, while SIT forms a clearly visible complex with S.