Arthrospira (Spirulina) platensis is known to have high-quality proteins content and phycocyanin as one of the major pigment constituents of the cells, and the most challenging problem associated with phycocyanin production in Arthrospira is to optimize its intracellular accumulation. The present study evaluated the metabolic stress conditions (by nutrient enrichment) of Arthrospira platensis FACHB-314 for boosting biomass growth and high content phycocyanin accumulation. Experimental results showed that 5 mM sodium glutamate and 7.5 mM succinic acid could enhance biomass yield as well as phycocyanin accumulation compared with that of the control groups. The present study demonstrates that the biomass growth and phycocyanin accumulation were significantly enhanced in fed-batch cultivation of Arthrospira platensis by applying the substrates as metabolic stress agents combined with nitrate feeding strategy. cobA/hemD, hemG and ho genes presented the over-expression level with adding sodium glutamate and succinic acid in cultures, respectively, compared to the control groups.