This work demonstrates the ability of the yeastYarrowia lipolyticacultivated on biodiesel waste to synthesize alpha-ketoglutaric acid with a minimal content of pyruvic acid as the main byproduct. The key factor promoting the microbial production of alpha-ketoglutaric acid from the waste is a strong deficiency of thiamine in the cultivation medium. The production of alpha-ketoglutaric acid by the yeast can be regulated by changing the concentration of nitrogen, iron, zinc, copper, and manganese in the medium, as well as by pH medium and the aeration rate. The optimization of these parameters in flask experiments allowed us to increase the concentration of alpha-ketoglutaric acid in the medium by 2.6 times and to shift the alpha-ketoglutaric acid/pyruvic acid ratio from 5:1 to 30:1. During cultivation in a fermentor under optimized conditions,Y. lipolyticaproduced 80.4 g/L alpha-ketoglutaric acid with a process selectivity of 96.7% and the product yield (Y-KGA) equal to 1.01 g/g.