There is a lack of fundamental knowledge about the scale up of biosurfactant production. In order to develop suitable technology of commercialization, carrying out tests in shake flasks and bioreactors was essential. A reactor with integrated foam collector was designed for biosurfactant production using Bacillus subtilis isolated from agricultural soil. The yield of biosurfactant on biomass (Y (p/x)), biosurfactant on sucrose (Y (p/s)), and the volumetric production rate (Y) for shake flask were obtained about 0.45 g g(-1), 0.18 g g(-1), and 0.03 g l(-1) h(-1), respectively. The best condition for bioreactor was 300 rpm and 1.5 vvm, giving Y (x/s), Y (p/x), Y (p/s), and Y of 0.42 g g(-1), 0.595 g g(-1), 0.25 g g(-1), and 0.057 g l(-1) h(-1), respectively. The biosurfactant maximum production, 2.5 g l(-1), was reached in 44 h of growth, which was 28% better than the shake flask. The obtained volumetric oxygen transfer coefficient (K (L) a) values at optimum conditions in the shake flask and the bioreactor were found to be around 0.01 and 0.0117 s(-1), respectively. Comparison of K (L) a values at optimum conditions shows that biosurfactant production scaling up from shake flask to bioreactor can be done with K (L) a as scale up criterion very accurately. Nearly 8% of original oil in place was recovered using this biosurfactant after water flooding in the sand pack.