Hyperthermophilic Clostridium sp. N-4 produced a glycoprotein biosurfactant that enhanced recovery of residual oil at 96 °C in lab studies.

Biosurfactant producing hypethermophilic microorganisms are essentially required for Microbial Enhanced Oil Recovery (MEOR) from high temperature oil reservoirs (above 90 °C). In the present study, biosurfactant producing Clostridium sp. N-4, optimally growing at 96 °C was isolated from a high temperature oil reservoir. Effect of pH, temperature and salinity on production and activity of N-4 biosurfactant was investigated. Biosurfactant produced by N-4 was partially purified by acid precipitation, characterized using FT-IR spectroscopy; and evaluated for its ability to enhance oil recovery in sand pack studies. The strain N-4 produced biosurfactant over a wide range of pH (5.0-9.0) and salinity (0-13%) at high temperature (80-100 °C) and optimally at pH 7, 96 °C and 4% salinity. N-4 biosurfactant was active at 37-101 °C; pH, 5-10 and salinity of 0-12 % (w/v). N-4 biosurfactant, characterized as glycoprotein reduced the surface tension of water by 32 ± 0.4 mN/m at critical micelle concentration of 100 μg/ml. N-4 biosurfactant mobilized 17.15% of residual oil saturation in sand pack studies. Similarly, the strain N-4 also recovered 36.92% of the residual oil in sand pack studies under the conditions mimicking the environment of depleted high temperature oil reservoir. Thus, the biosurfactant producing Clostridium sp. N-4 was identified as a suitable agent for enhanced oil recovery from high temperature oil reservoirs.