To promote the production of ?-ketoglutaric acid (?-KG) by Yarrowia lipolytica WSH-Z06 with glycerol as the carbon source, the effects of pH on cell growth and ?-KG production were compared, and a strategy of controlling biomass by pH regulation with analysis of kinetic characteristics was established. Furthermore, the effect of biomass concentration on ?-KG production was compared. The results showed that the pH value was maintained at 6.0 at beginning of fermentation, and decreased naturally to 3.0 after the biomass concentration reached up to 10 g/L, then maintained till the end of fermentation. Using this strategy, the maximum ?-KG production reached up to 54.0 g/L, the production and productivity were 19.3% and 17.8% higher than the corresponding values of control group, respectively.