Light plays a significant role in microalgae cultivation, significantly influencing critical parameters, including biomass production, pigment content, and the accumulation of metabolic compounds. This study was intricately designed to optimize light intensities, explicitly targeting enhancing growth, pigmentation, and antioxidative properties in the green microalga, Scenedesmus falcatus (KU.B1). Additionally, the study delved into the photosynthetic efficiency in light responses of S. falcatus. The cultivation of S. falcatus was conducted in TRIS-acetate-phosphate medium (TAP medium) under different light intensities of 100, 500, and 1000 μmol photons m−2·s−1 within a photoperiodic cycle of 12 h of light and 12 h of dark. Results indicated a gradual increase in the growth of S. falcatus under high light conditions at 1000 μmol photons m−2·s−1, reaching a maximum optical density of 1.33 ± 0.03 and a total chlorophyll content of 22.67 ± 0.2 μg/ml at 120 h. Conversely, a slower growth rate was observed under low light at 100 μmol photons m−2·s−1. However, noteworthy reductions in the maximum quantum yield (Fv/Fm) and actual quantum yield (Y(II)) were observed under 1000 μmol photons m−2·s−1, reflecting a decline in algal photosynthetic efficiency. Interestingly, these changes under 1000 μmol photons m−2·s−1 were concurrent with a significant accumulation of a high amount of beta-carotene (919.83 ± 26.33 mg/g sample), lutein (34.56 ± 0.19 mg/g sample), and canthaxanthin (24.00 ± 0.38 mg/g sample) within algal cells. Nevertheless, it was noted that antioxidant activities and levels of total phenolic compounds (TPCs) decreased under high light at 1000 μmol photons m−2·s−1, with IC50 of DPPH assay recorded at 218.00 ± 4.24 and TPC at 230.83 ± 86.75 mg of GAE/g. The findings suggested that the elevated light intensity at 1000 μmol photons m−2·s−1 enhanced the growth and facilitated the accumulation of valuable carotenoid pigment in S. falcatus, presenting potential applications in the functional food and carotenoid industry.