ABSTRACT:
The study were performed to extracted and purified pyoluteorin produced by P. aeruginosa PA3 isolated from rhizospheric plant Wheat, studying its effect on some urepathogenic bacteria and candida isolated from locally hospitals in Baghdad/Iraq. the isolates involvingEscherichia coli, Klebsilla pneumonia, Pseudomonas aeruginosa,proteus mirabilis Acinetobacter baumannii, Serratiamarcescens and Enterobacter cloacae (gram-positive bacteria including Streptococcus agalactiae Staphylococcus aureus (10 isolates) and Staphylococcus epidermidis and the yeast Candida albicans (3 isolates), all isolates were identified using macroscoping, microscoping, biochemical tests and VITEK 2 system were used to confirmed the identification.PA3was culturing on PPM (pigment production medium) and extracted crudepyoluteorin with ethyl acetate, purified it with gel filtration chromatography on silica gel column and exposure the active fraction to TLC (thin layer chromatography), Rf-value obtained was 0.5, then exposure to HPLC to detected thepyoluteorin on wave length 308nm. Single peak appeared on retention time 7.02, which related to pyoluteorin. FTIR showed the chemical structures and chemicals bound found in crude pyoluteorin .Antimicrobial activity of crude and purified pyoluteorin of 3different concentration (50,100,200µg/ml) were detected against MDR Gram-positive Streptococcus agalactiae, Staphylococcus aureus, Gram-negative Escherichia coli, Proteus mirabilis, Acinetobacter baumannii and Enterobacter cloacae yeast as Candida albicans. The result obtained the purified pyoluteorin on concentration of 200µg/ml had greater activity than crude . Biofilm production by Klebsilla pneumonia (12 isolates) and Staphylococcus aureus (10 isolates) were detected.Purified pyoluteorin could prevention the biofilms formation.