This study evaluated the growth and biochemical composition of farming Gracilaria gracilis (Stackhouse) M. Steentoft, L. M. Irvine & W. F. Farnham in the Bizerte Lagoon (BL) and Bizerte Bay (BB) in the North Coast of Tunisia, using lantern nets. Effects of site and depth on alga daily growth rate (DGR) and biochemical composition were investigated. The DGR was affected by culture site (1.42 ± 0.65% day-1 and 1.19 ± 0.34% day-1 for the BL and the BB respectively). Agar yield, was higher (p < 0.05) in the BB than the BL (23.31 ± 2.64% vs. 19.19 ± 2.32%) with a higher (p < 0.05) 3,6-anhydrogalactose (3,6-AG) contents (41.37 ± 3.68% vs 23.30 ± 5.40%) and a lower (p < 0.05) sulphate degree (6 ± 2.00% vs 8.80 ± 0.86%). The proteins contents were independent of the site and depth of culture (20.74 ± 7.22% and 22.02 ± 6.34% for the BL and the BB respectively). R-phycoerythrin (R-PE) contents were significantly higher (p < 0.05) in the BB (0.86 ± 0.31 mg g-1) than those obtained in the BL (0.33 ± 0.12 mg g-1). The salinity, transparency, nitrate and ammonium were monitored in both sites, and their influences were discussed. Our results suggest that G. gracilis cultured in Bizerte Bay can be used in a cascading biorefinery approach.