EMULSAN ANALYSIS PRODUCED BY LOCALLY ISOLATED BACTERIA AND ACINETOBACTER CALCOACETICUS RAG-1

Growth of previously isolated bacteria from Iranian oil reservoirs on different carbon and energy sources and under varying conditions have been used to produce a class of extracellular microbial protein-associated lipopolysaccharides named emulsan.Several Bacteria were previously isolated from Iranian oil reservoirs and designated as; Ilam-1 and Paydar-4. In present study, the isolated strains were compared with standard sample of Acinetobacter calcoaceticus RAG-1 from Persian Type Culture Collection (PTCC 1641), IROST. Among the isolated strains, two strains were found to produce an extracellular, emulsifying agent when grown in Mineral Salt Medium containing soya oil, ethanol or local crude oil. The isolated bacteria were cultured and further analysed using protein estimation, reducing sugar analysis, hemolytic activity, surface tension and emulsification activity tests. The crude emulsifier of RAG-1, PAYDAR-4 and ILAM-1 were concentrated from the cell-free culture fluid by ammonium sulfate precipitation to yield 1.89g, 1.78g and 1.69g of bioemulsan respectively. Emulsifying activity was observed over the entire production process. These investigations showed that emulsan produced by isolated Iranian crude oil reservoir were comparable with Acinetobacter calcoaceticus RAG-1 which is made of carbohydrate backbone as its hydrophilic part (N-acetyl-D-galactoseamine, N-acetylgalactoseamine uronic acid, diamino-6-deoxy-D-glucose) and fatty acid chain as its hydrophobic portion.