Antibacterial activity of the water-soluble N-alkylated disaccharide chitosan derivatives against Escherichia coli and Staphylococcus aureus was investigated. It was found that the antibacterial activity of chitosan derivatives was affected by the degree of substitution (DS) with disaccharide and the kind of disaccharide present in the molecule. Regardless the kind of disaccharide linked to the chitosan molecule, a DS of 30-40%, in general, exhibited the most pronounced antibacterial activity against both test organisms. E. coli and S. aureus were most susceptible to cellobiose chitosan derivative DS 30-40% and maltose chitosan derivative DS 30-40%, respectively, among the various chitosan derivatives examined. Although the disaccharide chitosan derivatives showed less antibacterial activity than the native chitosan at pH 6.0, the derivatives exhibited a higher activity than native chitosan at pH 7.0. Antibacterial activity of the chitosan derivatives (DS 30-40%) against E. coli increased as the pH increased from 5.0 and reached a maximum around the pH of 7.0-7.5. The effect of pH on the antibacterial activity of chitosan derivatives against S. aureus was not as pronounced as that observed with E. coli. Population reduction of E. coli or S. aureus in nutrient broth increased markedly upon increasing the concentration of chitosan derivatives from 0 to 500 ppm. No marked increase in population reduction was noted with further increase in the concentration of chitosan derivatives even up to 2000 ppm.