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, 61.3 g l(-1); specific rate of PA production, 76.3 mg PA (g cells h)(-1); mass yield coefficient (YPA), 0.71; and energy yield coefficient (eta(PA)), 0.53). A relationship between PA production and activities of enzymes involved in a primary glycerol oxidation, citrate and glyoxylate cycles (GCs) is discussed. (C) 2003 Elsevier Ltd. All rights reserved.