The main objective of this research was to determine the effect of salinity on growth, and production of bio-relevant molecules, from green microalgae: Oedogonium, Cladophora, Ulothrix, and Spirogyra using response surface methodology (RSM) together with central composite design (CCD). Investigated algal biomass decreased (4-–fivefold), whilst carbohydrates decreased ranging from (0.14–0.19 mg/g), proteins (28–36%), and chlorophyll by (17–50%) using a treatment 100 mM of NaCl. The reduction in carbohydrates, proteins, and pigments was accompanied with a significant increase in lipids ranging from 33 to 80% at 100 mM NaCl. Saturated fatty acids were increase from 34.3 to 49.5%, PUFAs from 12.0 to 27.9% while MUFAs decreased from 52.8 to 19.4% at 80 mM of NaCl. Biodiesel-related values including iodine value (less than 120 g I2/100 g), saponification value (161–209 mg/KOH/g), cetane number (greater than 47 except for Cladophora sp.), cold filter plugging point (− 13.84 to − 16.39 °C), density (0.86–0.88 gcm−3), kinematic viscosity (3.5–3.9 mm2/s), oxidative stability (46–382 h at 110 °C), and heating value (36 to 41 MJ/kg) were in the satisfactory ranges of biodiesel standards EN 14,214 & ASTM D6751-02. Our results indicate that the investigated green algal species can produce large lipid yields and biofuels using brackish, saline water in contrast to conventional and current systems.