The purpose of this study was to produce microbial oil from Yarrowia lipolytica Po1g grown in defatted rice bran hydrolysate. After removing oil from rice bran by Soxhlet extraction, the bran is subjected to acid hydrolysis with various sulfuric acid concentrations (1-4% v/v), reaction times (1-8 h), and reaction temperatures (60-120°C). The optimal conditions for maximum total sugar production from the hydrolysate were found to be 3% sulfuric acid at 90°C for 6 h. Glucose was the predominant sugar (43.20 ± 0.28 g/L) followed by xylose (4.93 ± 0.03 g/L) and arabinose (2.09 ± 0.01 g/L). The hydrolysate was subsequently detoxified by neutralization to reduce the amount of inhibitors such as 5-hydroxymethylfurfural (HMF) and furfural to increase its potential as a medium for culturing Y. lipolytica Po1g. Dry cell mass and lipid content of Y. lipolytica Po1g grown in detoxified defatted rice bran hydrolysate (DRBH) under optimum conditions were 10.75 g/L and 48.02%, respectively.