The pineapple substrate was collected from the market in Chidambaram. The materials were cut into small pieces and juice was extracted. The juice substrate were subjected to physical, chemical and biological methods of pretreatment in order to liberate the fermentable sugars from the complex polysaccharides to select efficient methods for pretreatment. The yeast Saccharomyces cerevisiae was isolated from the extracted pineapple juice. The growth rate of Saccharomyces cerevisiae cultures were tested by using different sugars and nitrogen source. Among these, glucose supported maximum growth of 0.74 % at 15 % concentration after 48 hours of incubation and among the nitrogen source, maximum cell density 1.47 was observed at 48 hours of incubation when 1000 mg of Ammonium sulphate was added. The fermentation parameters such as pH, temperature and inoculums size were optimized for enzyme hydrolyzed grape juice residues. The pH parameters are 3.5, 4.5, 5.5 and 6.5 was taken. Among these 5.5 were ideal for yeast fermentation. The ethanol yield obtained with optimized parameters was 22.17g L-1. The temperature level of 44 ºC and inoculums size of 1 % were ideal for yeast fermentation.