Scaled-up production and recovery of lipopeptide biosurfactant and its application for washing petroleum-contaminated drill cuttings

A simple and efficient fermentation strategy using chitosan-immobilized Bacillus subtilis GY19 was developed for large-scale lipopeptide production from waste glycerol and palm oil. When a fed-batch fermentation approach was employed, excessive foaming was minimized, while lipopeptide production was greater than that of conventional batch fermentation. When a 700 L stirred tank fermenter with a 400 L working volume was used, a lipopeptide concentration and yield of 7.55 g/L and 0.190 g/g of substrate, respectively, were achieved. The pilot-scale recovery process involved sequential bacterial cell removal via centrifugation, elimination of impurities through acidification of the cell-free broth, and partial purification and concentration of the lipopeptides via ultrafiltration. A 5 kDa stabilized cellulose-based membrane maintained a constant permeate flux and resulted in 67.2% lipopeptide recovery. The concentrated lipopeptide retentate contained 16.8 g/L lipopeptides and a surface tension of 28.82 mN/m. A biobased washing agent for the treatment of petroleum-contaminated drill cuttings was later formulated with the retentate and various vegetable oil-based surfactants via the simplex-lattice mixture design methodology. The mixture of 1 % retentate and 4% fatty alcohol ethoxylate 5EO presented the greatest removal efficiency of 83.2% for drill cuttings containing 15% (w/w) total petroleum hydrocarbons. The formulation also mobilized petroleum hydrocarbons into a distinct free oil layer, which would be easy to separate and subsequently recycle. These findings demonstrate the scalability of the lipopeptide production and recovery process, while the biobased washing agent offers an environmentally friendly approach for petroleum remediation.