Biosynthesis of pyocyanin pigment by Pseudomonas aeruginosa

Abstract

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 mg/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 þve bacteria than GramÀve 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%.

Isolation and characterization of nutrient dependent pyocyanin from Pseudomonas aeruginosa and its dye and agrochemical properties

“…Similar investigation was performed by E-Fouly et al 27 in which pyocyanin production from P.aeuroginosa was carried out using glycerol supplemented nutrient broth (GSNB), Mineral Medium and King's A broth and it was concluded that the production of pyocyanin was enhanced by using raw materials as nutrient sources. These results support our conclusion that the production of pyocyanin is nutrient dependent and the microbes have an advantage in synthesis of important pigments by the use of a variety of raw nutrients under varying cultivation processes 27,28 . Hence, based on our results it can be considered as a low cost chemical and natural nutrients are essential for the microbial growth and metabolism underlying the glycolysis pathway, where microbes used food for the energy to run cellular processes, thereby bestow enhanced production of pyocyanin.…”

Bioactivity of pyocyanin of Pseudomonas aeruginosa clinical isolates against a variety of human pathogenic bacteria and fungi species

“…The study of El fouly in Egypt, found that P. aeruginosa isolated from human urine sample produced more pyocyanin concentration than isolates from rice cultivated soil (production titer was 6.3 and 5.9 mg/ml, respectively) [19], While Ra'oof and Latif, in Iraq, has detected that the P. aeruginosa isolate from sputum produced the largest amount of pyocyanin (12.069 µg/ml) [29]. The study of Charyulu et al (2009), has shown that the secondary metabolite from Pseudomonas which played a significant role in lyses the pathogenic bacterial including MRSA strains [30].…”

“…While the study of Al-Imari in Baghdad/Iraq, has reported the incidence P. aeruginosa isolates from certain clinical specimens as follow: urine (22.4%), ear (18.4%), sputum (16.3%), burns (14.3%) and wounds (12.3%) [18]. El fouly and coworkers in Cairo, Egypt found that P. aeruginosa was the most common cause of urinary tract infection (12.5%), followed by burns (10%) and sputum (9%) [19]. The reasons of variations in the occurrence rates of P. aeruginosa infections may be due to underlining diseases of investigated patients, virulence factors and the local antimicrobial resistance of the organism, and other health conditions [2].…”

Plakofuranolactone as a Quorum Quenching Agent from the Indonesian Sponge Plakortis cf. lita

“…The experiments were performed in triplicate. Pyocyanin concentration (µL/mL) was calculated as: $\mathrm{normalP}=\left(\mathrm{OD}\times 17.072\right)\times 1.5$ where OD is optical density value obtained at 520 nm, 17.072 is the extinction coefficient used to obtain the value in µg/mL, and 1.5 is the dilution factor (3 mL from initial 4.5 mL of chloroform was used) [39]. …”