Trehalose-6-phosphate synthase activity was determined by colorimetric, spectrophotometric and trehalose specific assays. All methods gave comparable results thus confirming our previous findings (1) and those reported by Elander (2). Different strains and mutants of Saccharomyces were carefully re-investigated in relation with the recent claim made by Vandercammen et al. (3) that our spectrophotometric assay over-estimated the enzyme activity and that no differences exist between wild type and mutant strains. In this paper we also confirm the de-activation of the trehalose synthase complex in response to a "glucose signal", and present trehalose-6-phosphate synthase and trehalase activities in different strains measured during all phases of growth on glucose.