Crude water extracts from Australian wattle seed (Acacia victoriae Bentham) and their salt (ammonium sulphate)-precipitated fractions were analysed for trypsin and '-chymotrypsin (chymotrypsin) inhibitor activity using gel electrophoresis and spectrophotometric methods. Three different bands with molecular weight 30.20, 38.03 and 39.81 kDa were active with the 50% salt-precipitated fraction exhibiting highest activity and number of active bands. The same proteins also appeared to be responsible for both trypsin and chymotrypsin inhibitor activity. To establish conditions for the inactivation of these inhibitors, whole seed and uncoated cotyledon were subjected to different heat treatments. Moist heat treatment at 100oC for 30 seconds was sufficient to inactivate both protease inhibitors although trypsin inhibitor was found to be more heat resistant than chymotrypsin inhibitor. Soaking overnight before thermal treatment improved the trypsin inhibitor activity but enhanced the efficiency of thermal inactivation in both inhibitors.