This study investigated the extraction and purification of marine pyoverdine from Pseudomonas otitidis EGY-NIOF-A1, a fluorescent pigment analyzed for purity using reverse-phase HPLC. Structural characterization through ¹H NMR, ¹³C NMR, FT-IR, and XRD revealed aliphatic regions, hydroxyl and amide groups, and a partially crystalline structure, all of which are essential for metal chelation. Cytotoxicity tests confirmed its environmental safety. The potential of pyoverdine for Cr (VI) bioremediation was evaluated through batch experiments, which showed maximum adsorption at pH 1 and a Langmuir isotherm fit, with an adsorption capacity of 63.69mg/ g. These findings position pyoverdine as an eco-friendly, cost-effective agent for chromium remediation, with the potential to enhance water quality in both environmental and aquaculture systems.