SCREENING HEMOLYTIC ACTIVITY OF LUMINESCENT VIBRIOS WITH FURTHER HEMOLYSIN EXTRACTION AND MOLECULAR WEIGHT DETERMINATION

Background and Aim:Many pathogenic and non-pathogenic bacteria secrete various extracellular proteins. Among these proteins, hemolysin is an exotoxin which lyses erythrocyte membranes and exerts various roles in the infection process. In many cases, the pore-forming activity of hemolysin is not restricted to erythrocytes, but extends to a wide range of other cell types open up its potential medical applications. The aim of this study was to investigate the hemolytic activity of luminescent Vibrio strains and hemolysin characterization of selected strain.Methods:Hemolysin-producing Vibrio strains were screened by culturing them on blood agar medium. Then the most active Vibrio strains enabled to create highest diameter of greenish zone was selected and designated as Vibrio harveyi HFB18. In addition, the 96 well plate hemolysin assay was performed to select the strain with highest hemolysin production. The V. harveyi HFB18 hemolysin was extracted by ammonium Sulphate precipitation and Zymography method. Finally, SDS-PAGE method was used to measure the molecular weight of HFB18 hemolysin.Results:V. harveyi HFB18 showed highest hemolytic activity among 4 Vibrio strains that were investigated. The V. harveyi HFB18 hemolysin was extracted successfully and its lysis properties were confirmed. SDS-PAGE method showed that this bacterium is able to produce hemolysin with molecular weight of 52 kDaConclusion:Successful isolation and purification of hemolysin from a non-pathogenic bacterium is advantageous and its characterization could reveal novel properties that may be used in biomedicine studies.

کلیدواژه ها:

Vibrio harveyi ، hemolysin ، non-pathogenic ، luminescence