Introduction: Melanin is a high-molecular weight pigment produced through the oxidative polymerization of phenolic or indolic compounds and plays a perfect role in UV-light shielding, as well as in photoprotection. Among biopolymers, melanin is unique in many aspects. This study is designed to screen Production, extraction and characterizes of an extracellular melanin pigment from clinically isolated P. aeruginosa. Objective: The aim of the current study is isolation and diagnosis of P.aeruginosa using vitek-2 compact system and screening the ability to produce melanin and characterization of extracted melanin by UV-vis, FTIR, XRD and SEM. Materials and methods: the samples swab inoculated on cetrimide agar as selective media and incubated aerobically for 24 hours at 37°Cand used nutrient agar with nutrient broth supplement with 1% tyrosine for Screening for melanin production by P. aeruginosa isolates. Results: Four isolates P. aeruginosa were identified out of 109 specimens have ability to produce melanin pigment under specific medium and culture conditions. According to morphological, cultural, biochemical, and VITEK-2 characteristics, isolates were identified as P. aeruginosa. The results showed that the isolate of burn was greatest in production of melanin. Based on its solubility in organic solvents, the black pigment was identified as melanin and structurally and functionally characterized by UV-Vis absorbance and presence of various characteristic peaks that determined by FTIR analysis, and the morphology of the pigment surface was examined using SEM and XRD spectra analysis. Conclusion: Depending on the type of bacteria and the conditions of their culture, different melanin-producing bacteria produce different amounts of melanin. The primary factors for bacterial melanin production are L-tyrosine, energy source, pH, temperature and surface area for shaker agitation. In this study, results found that the optimum condition for melanin production byP. aeruginosa in pH=7.3 and the effect of L-tyrosine concentration on melanin production found that use 1g/L is the best concentration for production.