The relationship between the concentration of intracellular ATP and the glycolytic flux in Torulopsis glabrata was studied by adding oxidative phosphorylation inhibitors (rotenone, antimycin A and oligomycin). When 10 mg/L rotenone and antimycin A were added to the cell cultures, the concentrations of intracellular ATP were approximately 43% and 27.7% less than that of the control, respectively. The specific activity of phosphofructokinase, one of the rate limiting enzymes of the glycolytic pathway, increased by a factors of 3.4 and 2.3, in comparison of the control respectively. With the specific activity of phosphofructokinase increased, the rate of glucose consumed increased by a factor of 3.6 and 2.4 compared with the control, and the rate of pyruvate produced increased by 17% and 8.5% respectively. The specific activities of hexokinase and pyruvate kinase were not affected by the addition of rotenone or antimycin A. Furthermore, the concentration of intracellular ATP decreased by 64.3% upon addition of 0.05 mg/L oligomycin to the cell culture, and the growth of Torulopsis glabrata was ceased when 0.4 mg/L oligomycin was added to the culture broth at 24 h. Both the rate of glucose consumed and the rate of pyruvate produced were enhanced with increasing the concentration of oligomycin ( ＜ 0.6 mg/L) in the cell cultures. As a result, the activity of phosphofructokinase (r²=0.9971), the rate of glucose consumed (r²=0.9967) and the rate of pyruvate produced (r²=0.965) were enhanced by a decrease in the energy level of the cell (the concentration of intracellular ATP). Increase of the rate of glucose consumed rooted in elevation of the specific activity of phosphofructokinase (r²=0.9958) and pyruvate kinase (r²=0.8706). These results are the first answer to the fundamental question of what controls the flux through glycolysis in Torulopsis glabrata.