The role of amylase present in honey in enhancing its antibacterial activity was evaluated in the presence and absence of starch. Two strains of pathogenic bacteria have been used: Staphylococcus aureus and Escherichia coli. For S. aureus, the minimum inhibitory concentration (MIC) for the three varieties of honey tested without starch was 11%, 24%, and 29% (vol/vol), respectively. When starch was added with honey to the media the MIC obtained was 5%, 19%, and 25% (vol/vol), respectively. For E. coli, the MIC for the three varieties without starch was 23%, 28%, and 25% (vol/vol), respectively. When starch was added with honey to media, the MIC was 19%, 26%, and 23% (vol/vol), respectively. It is speculated that the amylase present in honey hydrolyzed the starch chains to randomly produce dextrin and maltose and that this increased the osmotic effect of the media, which consequently increased the antibacterial activity.