One-step purification and characterization of alginate lyase from a clinical Pseudomonas aeruginosa with destructive activity on bacterial biofilm

One-step purification and characterization of alginate lyase from a clinical Pseudomonas aeruginosa with destructive activity on bacterial biofilm

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Objective(s): Pseudomonas aeruginosais a Gram-negative and aerobic rod bacterium that displays mucoid and non-mucoid phenotype.Mucoid strains secrete alginate, which is the main agent of biofilms in chronic P.aeruginosa infections, show high resistance to antibiotics; consequently, the biological disruption of mucoid P.aeruginosa biofilms is an attractive area of study for researchers.Alginate lyase gene (algl) is a member of alginate producing operon which by glycosidase POTION 9 activity produces primer for other enzymes in this cluster.

Also this activity can destroy the extracellular alginate; therefore this enzyme participates in alginate production and destruction pathway.Alginate lyase causes detachment of a biofilm by reducing its adhesion to the surfaces, and increases phagocytosis and antibiotic susceptibility.In this study, alginate lyase was purified in just one step Chaste Berry ( Vitex ) and its properties were investigated.Materials and Methods: The purification was done by affinity chromatography, analysed by SDS-PAGE, and its effect on P.aeruginosa biofilms was surveyed by micro titer plate assay and SEM.

The substrate specificity of the enzyme was determined by PCR.Results: Alginate lyase from isolate 48 was purified in one step.It is more thermally resistant than alginate lyase from Pseudomonas aeruginosa PAO1 and poly M, poly G and poly MG alginate were the substrate of this enzyme.Moreover, it has an eradication effect on biofilms from P.aeruginosa 48 and PAO1.

Conclusion: In this study an alginate lyase with many characteristics suitable in medicine such as thermal stability, effective on poly M alginate, and bacterial biofilm destructive was introduced and purified.