Algal growth limitation in large-scale cultivation mostly results from high level synthesis of photosynthetic pigments, owing to self-shading effects and attenuation of light distribution. To overcome this problem, here we investigated the influence of nitrogen modulation on changes in antenna pigments as well as biomass and lipid production by Chlorella vulgaris under a chemostat continuous cultivation mode. The production of algal antenna pigments, including chlorophylls and carotenoids, was promoted in a total nitrogen (TN) concentration-dependent manner. Maximum algal biomass and lipid production were obtained from 70 mg/L of TN concentration along with a significant increase in light transmittance and reduction in antenna pigments. Furthermore, the composition of polyunsaturated fatty acids remarkably augmented at low TN concentrations. These results suggest that the reduction in algal antenna pigment synthesis via modulation of nitrogen concentration may serve as an effective strategy to enhance algal biomass and lipid production.