The bioethanol production from sugar beet is regulated by endophytic bacteria. Thus, the present investigation is aimed to isolate, identify and characterize endophytic bacteria from different varieties of sugar beet roots. The presence of specific bacteria in sugar beet roots plays vital role in bioethanol production. The ability to produce bioethanol through fermentation with the help of various microbes present in roots was studied in different varieties of sugar beet at Indian Institute of Sugarcane Research, Lucknow sub tropical regions of India. A total of 16 microbial isolates comprising of 12 bacterial and 4 fungal strains were studied for their potential to produce bioethanol from sugar beet root. Among them only 2 bacteria were more effective as fermenting agents. These isolated endophytic bacteria were characterised by molecular sequencing and were identified as Enterobacter hormaechei and Enterobacter cloacae. These isolates may also act as a good alternative agent for the bioethanol production.