Enhancement of glycerol production in Shochu yeast, which was induced by heat-shock treatment, was studied. Although heat-shock treatment (45 degrees C, 1 h) caused a transient delay in cell growth, the amount of glycerol produced by heat-shock-treated cells was 20% higher than that by control cells. During the glycerol-production phase, the NAD(+)-dependent glycerol-3-phosphate dehydrogenase (GPDH) activity of heat-shock-treated cells was much higher than that of control cells, suggesting that a higher GPDH activity enhances glycerol production. The level of NAD(+)-dependent glycerol dehydrogenase (GDH) activity was almost the same between heat-shock-treated cells and control cells. The results of Northern blot analysis of GPD genes (GPD1 and GPD2) encoding the GPDH enzyme showed that the transcription of GPD genes was not affected by heat-shock treatment but the period of intensive transcription of GPD1 was prolonged.