This study explores the utilization of a novel microbial green surfactant (lipopeptide surfactin), to facilitate the pulse electrodeposition of brushite (CaHPO4·H2O) and hydroxyapatite (Ca10(PO4)6(OH)2) coatings on stainless steel (316L SS). The addition of the biosurfactant and change in the current density (CD) influence the coating composition and its morphology. Characterization techniques such as X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy are employed to analyze the coatings composition. In the absence of biosurfactant, the coatings primarily consist of brushite phase with a minor content of hydroxyapatite. However, with the addition of biosurfactant as an additive, the brushite phase becomes the prominent constituent, suggesting the biosurfactant’s inhibitory effect on the growth of the hydroxyapatite phase. Scanning electron microscopy (SEM) analysis reveals distinct flower-like morphology in the biosurfactant-assisted coatings, while the other coatings exhibit a broom-like morphology. Additionally, the incorporation of the biosurfactant leads to a reduction in crystallite size, an increase in surface roughness, and improved adhesion strength of the coatings. To assess cytocompatibility, the MTT assay is performed using fibroblast cells (L929). SEM analysis evaluates cell adhesion, morphology, and interactions with the coatings. Furthermore, the addition of biosurfactant to the electrolytic solution also results in improved corrosion resistance. Overall, these findings highlight the potential of biosurfactant-aided brushite-coated surfaces for applications in bone regeneration, tissue engineering, and biomedical fields.