The production of pigments by Haloferax mediterranei ATCC 33500 with two-stage cultivation in response to nutritive factors in culture media was studied. Sodium chloride and magnesium sulfate in the second-stage media showed a marked effect upon the production of pigments, and sodium acetate could enhance the production. As the cells were harvested at mid-log phase of growth in first-stage cultivation and transferred to the defined media containing 5% sodium chloride, 0.1% sodium acetate and 8% magnesium sulfate at 37°C, 120rpm for further 24h of cultivation, H. mediterranei exhibited to be an efficient producer of pigments. The yield of pigments could reach up to 0.604A 494nm mL −1 broth. TLC analysis and the UV–Vis spectra of individual spots thereof revealed that H. mediterranei produced three red pigments of C 50 carotenoid, namely bisanhydrobacterioruberin, monoanhydrobacterioruberin and bacterioruberin, as well as a C 45 carotenoid, 2-isopentenyl-3,4-dehydrorhodopin.