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This study investigated the separation of 1,3-propanediol from a fermentation broth produced using Klebsiella pneumoniae. Salting-out extraction was used to separate 1,3-propanediol using one-salt and two-salt systems. The highest distribution coefficient and recovery of 1,3-propanediol between pentanol and an aqueous solution of a single-salt system using sodium phosphate (0.4 g/mL) were 3.72% and 72%, respectively; and 2.52% and 65.6%, respectively, when using sodium sulfate (0.4 g/mL). For the two-salt system using sodium phosphate (0.4 g/mL) and sodium sulfate (0.2 g/mL), the results were 47.2% and 92.5%, respectively. The stripping recovery of 1,3-propanediol using pure water was approximately 90%.