Biosurfactants have recently gained popularity because they have numerous benefits over chemical synthetic surfactants, including higher biodegradability, lower toxicity, higher foaming, environmental compatibility, and effective properties under harsh conditions. This study aimed to produce effective biosurfactants by selected bacterial strains isolated from Egyptian oil fields to improve oil recovery and investigate their environmental aspects for microbial enhanced oil recovery. The selected strains were incubated in a new proposed nutrient medium H to produce biosurfactants with optimum surface and emulsification activities. Stability studies were conducted to examine the tolerance of produced biosurfactants in harsh reservoir conditions. Core flooding tests were performed to investigate the potential of produced biosurfactants in enhancing oil recovery. The environmental risk assessment was conducted to investigate if there are any possible threats of the selected bacterial strains. Results showed that selected bacterial strains Bacillus licheniformis and Bacillus subtilis could produce effective biosurfactants that reached their maximum surface activity after 24 h of incubation by reducing the surface tension from 71.8 mN/m to 27.13 mN/m and 25.74 mN/m, and the interfacial tension against kerosene from 48.4 mN/m to 1.27 mN/m and 0.38 mN/m at critical micelle concentration of 0.06 g/l and 0.04 g/l, respectively. The produced biosurfactants by Bacillus licheniformis and Bacillus subtilis showed significant emulsification activity against crude oil with emulsification indices of 50.2% and 63.7%, respectively. High stability was observed at high temperatures for a long-time period and more than 60% of their surface and emulsification activities were maintained over a wide range of pH and salinity. It was also found that 31.41–39.35% of additional oil could be recovered by the produced biosurfactants. Finally, Bacillus licheniformis and Bacillus subtilis are environmentally safe, have no potential for toxicity, and no risk could occur for MEOR.
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