A bio-floatation technique was used to test the effectiveness of Jatropha curcas seed extract in recovering microalgae from an aqueous medium. This technique used Jatropha curcas oil-protein extract emulsion for floatation removal of Chlorella vulgaris microalgae. The effects of variables have been investigated in this process, including operating parameters such as protein-oil emulsion dosage, pH, and mixing time. A maximum floatation efficiency of 81% was observed when a 20 ml/l protein-oil emulsion dosage was tested at pH 2 and a mixing time of 4 min. Furthermore, the analysis of the microalgae’s zeta potential changes revealed a notable variation in value caused by the protein-oil emulsion. This bio-floatation technique is useful for the high-efficiency harvesting of microalgae to produce cost-effective biofuel.