Background: Rhodotorula is characterized by the absence of ballistoconidia, fermentationability, and starch-like compounds. Biology of the species is not well-identified; thereforemolecular identification is required. Sequence analysis of the D1/D2 region can be usedfor the identification of the majority of Basidiomycetous species. carotenoids which arenatural pigments can be synthesized by some genera of yeasts such as Rhodotorula. Theincrease of demand for carotenoids obtained from natural sources has promoted majorefforts to recognize potential microbial sources. The aims of this study were to identify astrain isolated from leather wastewater and to investigate its carotenoid production ability.The effect of 2 different medium (Semi-synthetic medium (MMS) and yeast malt extractmedium (YM)) on biomass and carotenoid production was studied.Materials and Methods: In this experimental study, sequence analysis of the D1/D2region in addition to morphological and biochemical characterization to identify the strainwas carried out. To isolate the carotenoid pigment, cells were suspended in acetone andbroken using a homogenizer, followed by centrifugation and supernatant was separated; thus pigments were measured spectrophotometrically at 450 nM using the extinctioncoefficient E1%450=2500.Results: Identification processes represented strain SG006 as a Rhodotorula slooffiae. Thesequence was deposited in the Gene Bank database with accession number JX997835. Theresults showed that SG006 are able to produce carotenoid and MMS medium promotedcarotenoid production.Conclusion: We found that Rhodotorula slooffiae showed the ability to producecarotenoid. However, further work is needed to optimize of the amount of product and tocharacterize the carotenoids.