The fermentative production of biosurfactant by Bacillus licheniformis R2 strain in bench-scale bioreactor under different dissolved oxygen tensions was investigated. The statistically optimized minimal media was used for biosurfactant production, where glucose was used as a carbon source. The batch fermentations were carried out at different concentrations of DO - 30%, 50%, 70%, and initial 100% (with no further control thereafter) at 30 °C and pH 6.5-7.2. In 30, 50 and 70% DO experiments, it was maintained during the course of the fermentation, using cascading mode with agitation. Adjusting the initial dissolved oxygen to 100% saturation, without any further DO control and collection of foam and recycle of biomass gave higher biosurfactant production. B. licheniformis R2 produced biosurfactant, thus reducing the surface tension and interfacial tension to 28mN/m and 0.5mN/m respectively in less than 10 hours. The results are indicative of the potential of the strain for the development of scaled-up biosurfactant production.