The aim of this study was to explore the relationship of intracellular glutathione with various oxidative stress markers and the stress protectant marker trehalose. In the first group of yeast cells, diethyl maleate was used for depletion of glutathione. A second group of yeast cells were incubated with amino acids constituting glutathione (GIu, Cys, Gly) to increase glutathione level. Increased level of oxidative stress marker like ROS, protein carbonyl formation and lipid peroxidation and decreased viability in glutathione-depleted cells were observed in the present study. The increased activity of antioxidant enzymes SOD and CAT in the glutathione depleted group suggests the interaction of different antioxidant defence system in Pachysolen tannophilus. Furthermore, the increased levels of trehalose in glutathione-depleted group shows that trehalose acts as a stress reducer in glutathione depleted Pachysolen tannophilus.