Azurin reported as a potential anticancer protein against breast cancer cell lines, evoked the researchers of novel methods for enhanced synthesis of azurin has initialized. Early researchers revealed the synthesis of azurin from different microbial sources specifically from Pseudomonas aeruginosa (P. aeruginosa). This study is concerned of azurin synthesis from different strains of P. aeruginosa with lucid homogeneity by customized methods. The growth of different P. aeruginosa strains 1934, 741, 2453, and 1942 for the synthesis of azurin were scrutinized for enhanced azurin synthesis. The enhanced azurin synthesis from P. aeruginosa strains was improved by the copper sulphate (CuSO4) and potassium nitrate (KNO3) containing media under facultative anaerobic condition. The purification and synthesis of azurin process was performed in Ion- exchange and gel- filtration chromatography. Molecular mass and secondary structure of azurin was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis circular dichroism (CD) spectroscopy to establish the purity of azurin from the above strain. High yield was reported in P. aeruginosa 2453 strain than other strains.