The present study was aimed to investigate the potential of Neem tree leaves (Azadirachta indica) in bioethanol production by using Saccharomyces spp. and Bacillus spp.as fermenting organisms. Dried powdered leaves of neem tree (Azadirachta indica) were hydrolyzed using different concentrations of 5%, 10%, 15% and 20% sulfuric acid and also 1%, 2%, 5% and 10% sodium hydroxide. The hydrolyzed samples were then fermented using Saccharomyces spp. and Bacillus spp. After fermentation the broths formed were distillated to obtain ethanol. Acidified potassium dichromate was used to determine the bioethanol produced. Further, the quantification of produced bioethanol was carried out using UV spectrophotometer and the bioethanol produced was compared to check whether Saccharomyces spp. or Bacillus spp. produced more e ethanol. It was found that Saccharomyces spp. was producing more ethanol i.e. 6% when neem leaves were treated with 1% NaOH.