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During the last two decades, fermentation process was getting more attention for the production of amino acids. It mainly revolves around production of essential amino acids which cannot synthesize by human. Present project was also based on isolation and screening of twenty one bacterial isolates from soil and water and tested their potential to ferment different media for the production of different amino acids. However, the main emphasis was given to isoleucine. As a whole, fifteen isolates produced isoleucine i.e., 70.0% while six appeared to be non producers for isoleucine. Out of eleven isolates obtained from soil, eight were producers and three non producers displaying 72.0% production level. This percentage remained almost the same in bacteria obtained from water.