BACKGROUNDGlobal food loss and waste have raised environmental concerns regarding the generation of greenhouse gases (e.g., carbon dioxide and methane gas), which directly contribute to climate change. To address these concerns, the present research aims to upcycle food waste into an alternative culture medium for the cultivation of microalgae. Various parameters including pretreatment of food waste (i.e., autoclave and non‐autoclave), concentration of food waste culture medium (i.e., 10%, 30%, 50%, 70%, 90% and 100%), harvesting efficiency and biochemical compounds of Chlorella sp. microalgae were carried out.RESULTSBased on the preliminary findings, the highest biomass concentration obtained from 10% food waste culture medium in the autoclave for Chlorella sp., including strains FSP‐E, ESP‐31 and CY‐1, were 2.869 ± 0.022, 2.385 ± 0.018 and 0.985 ± 0.0026 g L−1, respectively. Since Chlorella vulgaris FSP‐E exhibited the highest biomass concentration, this microalgal strain was selected to examine the subsequent parameters. Cultivation of C. vulgaris FSP‐E in 100FW achieves a biomass concentration of 4.465 ± 0.008 g L−1 with biochemical compounds of 6.94 ± 1.396, 248.24 ± 0.976 and 406.23 ± 0.593 mg g−1 for lipids, carbohydrates and proteins, respectively.CONCLUSIONThis study shows that using food waste as an alternative culture medium for C. vulgaris FSP‐E can achieve substantial biomass productivity and biochemical content. This research work would contribute to the concept of net zero emission and transitioning toward a circular bioeconomy by upcycling food waste as an alternative culture medium for the cultivation of microalgae. © 2024 Society of Chemical Industry.