The present study investigated the effects of different phenol concentrations (200 – 1000 mg L−1) towards Chlorella sp. under different culture conditions (light vs. dark) and NaNO3 concentrations (0 – 0.1 g L−1) using central composite design. Phenol induced hormesis effects on the algal growth and cellular metabolites. Nitrate was identified as a crucial factor for promoting the uptake of phenol by Chlorella cells, while light was a limiting factor for growth, but the phyco-toxicity of phenol was decreased in the dark. The pigment contents were generally increased in the treated cells to protect against the oxidative phenol stress. The incorporation of 200 mg L−1 phenol and 0.05 g L−1 NaNO3 to the illuminated cells markedly promoted biomass and lipid contents to 0.22 g L−1 and 26.26% w/w, which was 44 and 112% higher than the phenol-less control, respectively. Under the same conditions, the increase of phenol concentration to 600 mg L−1, the protein contents were increased to 18.59% w/w. Conversely, the algal cells were able to accumulate more than 60% w/w of soluble carbohydrates under dark conditions at 600 mg L−1 of phenol. Nitrate replete conditions stimulated lipid accumulation at the expense of protein biosynthesis. Furthermore, most of the treatments showed an increase of H2O2 and malonaldehyde contents, especially for the illuminated cells. However, catalase activity tended to increase under dark conditions, especially at low phenol and nitrate concentrations. This study is valuable in indicating the effects of phenol on microalgae by exploiting response surface methodology, which can be applied as a powerful tool in growth monitoring and toxicity assessment.