The effects of the concentrations of inorganic salts in Schenk and Hildebrandt (SH) medium on catharanthine production in hairy root cultures of Catharanthus roseus were investigated. The inorganic salt components could be categorized into four groups. The first group (nitrate) supported both the growth of and catharanthine production by hairy roots with incremental increases in the concentration. The second (ammonium and phosphate) yielded contradictory effects with respect to growth and production. The third (borate and molibdate) inhibited both growth and production, while the fourth (potassium iodide, sulfate, and iron) did not exhibit any significant effects. Through optimization of the concentrations of inorganic salts in the medium, a two stage process of hairy root cultures with different media for growth and production was developed which enabled us to enhance the volumetric yield of catharanthine up to 60.5 mg/l. This productivity was 5.4 times higher than that of a one stage culture in the original SH medium.