The influence of heat shock induced during the exponential growth phase of the Saccharomyces cerevisiae NCIM 3095 and NCIM 3287 fermentation of Guava juice were examined. A rapid temperature change from 20°C to 38°C was applied and gives maximum glycerol and ethanol production. The effect of the duration of exposure to a high temperature has been analyzed. By the influence of a heat shock treatment, glycerol production was increased by up to 28% and 44.4 % respectively in NCIM 3095 and 3287 in guava wine production. This is a noble method for high glycerol production in large scale guava wine production.