Dandekar & Chandler:Pyocyanin Extraction

Pyocyanin Extraction

Pyocyanin [1] is a secondary metabolite secreted by the Gram negative bacterium Pseudomonas aeruginosa[2]. This protocol was established to quantify the pyocyanin produced by a P. aeruginosa culture in broth media.

Principle

Pyocyanin is an extracellular redox-active phenazine that contains a distinctive blue-green pigment. This compound can be isolated from cellular contaminants by isolating it in an aqueous suspension formed by the addition of chloroform. Because of the unique pigmentation this compound can then quantified with a spectrophotometer.

Materials Needed

• Spectrophotometer capable of reading 520 nm
• Vortex
• Sterile Cuvets
• Nanopure H20
• 10mL test tube
• 0.2m HCl
• Chloroform
• 1.5ml eppendorf tubes