Saccharomyces cerevisiae contains three chitin synthetases, Chs1, Chs2, and Chs3, performing different physiological functions but catalyzing the same reaction. It has been found that Ni2+ is a powerful inhibitor of Chs1 and Chs2 activity but has very little effect on the activity of Chs3, especially in the presence of Co2+. These results, together with the previous knowledge that Co2+ stimulates Chs2 and Chs3 but inhibits Chs1 and that the three synthetases differ in their pH optimum, have enabled us to formulate conditions for the specific determination of each synthetase in the presence of the others.