Background. Bacterial contamination is an ongoing problem for commercial bioethanol plants. It concerns factories using grain and also other raw materials for ethanol fermentation. Bacteria compete with precious yeasts for sugar substrates and micronutrients, secrete lactic and acetic acids, which are toxic for yeast and this competition leads to significant decrease of bioethanol productivity. For this study, bacterial contamination of corn grain was examined. Then the grain was treated by ammonia solution to reduce microbial pollution and after that the microbiological purity of grain was tested one more time. Disinfected and non-disinfected corn grains were ground and fermentation process was performed. Microbiological purity of this process and ethanol yield was checked out. Material and methods. The grain was disinfected by ammonia solution for two weeks. Then the grain was milled and used as a raw material for the ethanol fermentation. The fermentation process was carried out in 500-ml Erlenmeyer flasks. Samples were withdrawn for analysis at 0, 24, 48, 72 hrs. The number of total viable bacteria, lactic acid bacteria, acetic acid bacteria, anaerobic bacteria and the quantity of yeasts and moulds were signified by plate method. Results. Ammonia solution effectively reduces bacterial contamination of corn grain. Mash from grain disinfected by ammonia contains less undesirable microorganisms than mash from crude grain. Moreover, ethanol yield from disinfected grain is at the highest level. Conclusions. The ammonia solution proved to be a good disinfection agent for grain used as a raw material for bioethanol fermentation process.