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Light (wavelength, intensity, and light/dark cycle) have been considered as one of the most important parameters for microalgae cultivation. In this paper, the effect of medium frequency intermittent light on <i>Auxenochlorella pyrenoidosa</i> (formerly <i>Chlorella pyrenoidosa</i>) cultivation was investigated. Three parameters of intermittent light, light intensity, light/dark ratio, and light/dark cycle were employed and the influence of these parameters on the productivity of <i>Auxenochlorella pyrenoidosa</i> was studied. The biomass yield and growth rates were mainly affected by the light fraction and cycle time. Light with 220 μE m⁻² s⁻¹ light intensity was determined as the optimal light intensity for biomass production. At the light intensity of 420 μE m⁻² s⁻¹, the results indicated that the intermittent light improved the biomass production with larger light/dark ratio compared with the continuous light. At a lower mean light intensity over time, the intermittent light should be more suitable for biomass growth and the decrease in the light/dark ratio (L/D) will lead to a higher biomass productivity. The light/dark cycle time has little influence on the biomass yield.