Response surface methodology (RSM) based on the 23 factorial central composite design (CCD) was employed to evaluate optimum culture conditions (temperature, light irradiance and agitation) to enhance biomass and phycocyanin content of Spirulina platensis. The predicted maximum biomass and phycocyanin content by RSM was 1.06 g L-1 and 107 mg L-1, respectively, whereas maximum biomass and phycocyanin content of 1.32 g L-1 and 127 mg L-1 was obtained in the validation experiments under optimized conditions after 10 days of cultivation. Further, influence of optimized conditions (temperature 33±2 ºC, light irradiance 44 μmol photons m-2 s-1 and a flow rate of 2.5 L min-1) on growth and phycocyanin content of S. platensis in 7L Panel photobioreactor (PPBR) cultivation was investigated. A 15 days production was carried out and it was observed that a maximum biomass yield of 2.42 g L-1 with a specific growth rate 0.202 day-1 and phycocyanin content of 228 mg L-1 was obtained in the PPBR. The optimum culture conditions obtained through response surface methodology were successfully determined to maximize the biomass and phycocyanin.