The thiamine-auxotrophic yeast Yarrowia lipolytica 374/4 was selected as a promising producer of pyruvic acid (PA). The thiamine concentration in the medium was shown to have a crucial effect on PA production by this strain. Under limitation of yeast growth by thiamine, the cell requirement in thiamine for PA production depended on a source of carbon and energy used. Cultivation conditions were established ensuring intense PA production by glycerol-grown yeast Y. lipolytica 374/4 (PA concentration, 61.3gl - 1 ; specific rate of PA production, 76.3mg PA (gcellsh) - 1 ; mass yield coefficient (Y P A ), 0.71; and energy yield coefficient (η P A ), 0.53). A relationship between PA production and activities of enzymes involved in a primary glycerol oxidation, citrate and glyoxylate cycles (GCs) is discussed.