Characterization of Pseudomonas sp. TMB2 produced rhamnolipids for ex-situ microbial enhanced oil recovery

The present study describes the ex-situ production of a biosurfactant by Pseudomonas sp. TMB2 for its potential application in enhancing oil recovery. The physicochemical parameters such as temperature and pH were optimized as 30 °C and 7.2, respectively, for their maximum laboratory scale production in mineral salt medium containing glucose and sodium nitrate as best carbon and nitrogen sources. The surface activity of the resulting culture broth was declined from 71.9 to 33.4 mN/m having the highest emulsification activity against kerosene oil. The extracted biosurfactant was characterized chemically as glycolipid by Fourier-transform infrared spectroscopy and ¹H and ¹³C nuclear magnetic resonance spectroscopy analyses. The presence of mono-rhamnolipids (Rha-C_8:2, Rha-C₁₀, Rha-C₁₀-C₁₀, and Rha-C₁₀-C_12:1) and di-rhamnolipids (Rha-Rha-C₁₂-C_10, Rha-Rha-C₁₀-C₁₀, and Rha-Rha-C₁₀-C_12:1) congeners were determined by liquid chromatography-mass spectroscopy analysis. The thermostability and degradation pattern of the candidate biosurfactant were tested by thermogravimetry assay and differential scanning calorimetry studies for its suitability in ex-situ oil recovery technology. The rhamnolipid based slug, prepared in 4000 ppm brine solution reduced the interfacial tension between liquid paraffin oil and aqueous solution to 0.8 mN/m from 39.1 mN/m at critical micelle concentration of 120 mg/L. The flooding test was performed using conventional core plugs belonging to oil producing horizons of Upper Assam Basin and recovered 16.7% of original oil in place after secondary brine flooding with microscopic displacement efficiency of 27.11%.