Biosynthesis of pyocyanin pigment by Pseudomonas aeruginosa

Sixty-three isolates belonging to the genus Pseudomonas were isolated from different environmental sources including; soil, water and clinical specimens. Twenty out of them were identified as Pseudomonas aeruginosa and individually screened for pyocyanin production. P. aeruginosa R 1; isolated from rice-cultivated soil and P. aeruginosa U 3 selected from clinical specimen (Urinary tract infection) were the highest pyocyanin producers; pyocyanin production reached 9.3 and 5.9 μg/ml, respectively on synthetic glucose supplemented nutrient medium (GSNB). The identification of both selected strains ( P. aeruginosa R 1 and P. aeruginosa U 3 ) was confirmed by 16S rRNA, the similarity with other strains available in database was 97% (with P. aeruginosa FPVC 14) and 94% (with P. aeruginosa 13.A), respectively. P. aeruginosa R 1 and P. aeruginosa U 3 are accessed at gene bank with accession numbers KM924432 and KM603511, in the same order. Pyocyanin was extracted by standard methods, purified by column chromatography and characterized by UV-Vis absorption, mass spectrometry and nuclear magnetic resonance. The antimicrobial activity of purified pyocyanin against multi-drug resistant microbes was investigated; the efficiency of pyocyanin was more obvious in Gram +ve bacteria than Gram−ve bacteria and yeast. To reduce the cost of pyocyanin production, a new conventional medium based on cotton seed meal supplemented with peptone was designed. The pyocyanin production of both selected strains P. aeruginosa R 1 and P. aeruginosa U 3 using the new medium is increased by 30.1% and 17.2%, respectively in comparison with synthetic GSNB medium, while the cost of production process is reduced by 56.7%.