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Rhamnolipid is the main group of biosurfactants predominantly produced by Pseudomonas aeruginosa, a ubiquitous and opportunistic pathogen, which limits its large-scale exploitation. Thus, cost-effective rhamnolipid production from a newly isolated nonpathogenic Enterobacter sp. UJS-RC was investigated. The highest rhamnolipid production (4.4 ± 0.2 g/L) was achieved in a medium constituting agroindustrial wastes (sugarcane molasses and corn steep liquor) as substrates. Rhamnolipid exhibited reduced surface tension to 72-28 mN/m with an em