Aureobasidium puliulans possesses the capacity for simultaneous formation of fructosyltransferase and glucosyltransferase in various sugar media including sucrose, maltose, glucose and fructose. Among them, sucrose (300 g/1) was the most suitable carbon source for fructosyltransferase production, while fructose (100 g/1) gave the maximal production of glucosyltransferase. There existed a critical concentration for the optimal formation of enzymes in sucrose, glucose and fructose media. By contrast, no effect of maltose concentrations up to 300 g/1 was observed. The specific activity of the glucosyltransferase on maltose medium was highest during the early period of fetmentation, after which a sharp decrease occurred, whereas fructosyltransferase activity on sucrose medium maintained a nearly constant rate for a given culture period. Concomitant production of fructosyltransferase and glucosyltransferase was investigated with different combinations of lower concentrations of sucrose and maltose. Maltose supplementation in sucrose media and sucrose addition to maltose media enhanced the activity ratios of fructosyltransferase to glucosyltransferase as compared to that of non-supplemented media. Several polymers and surfactants were added in an attempt to enhance enzyme production, and supplementation of polyoxyethylene-sorbitan monolaurate (Tween 20) promoted fructosyltransferase production by 20%.