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Staphylococcus aureus (S. aureus) is one of the prominent gram positive human pathogensecretes many surface and secretary proteins including various enzymes and pathogenic factors that favourthe colonization and infection of host tissue. Alpha-amylase is one of the enzymes secreted by S.aureuswhich catalyses the breakdown of complex sugars to monosaccharides, which are required for colonizationand survival of this pathogen.Materials and Methods: In the present study we have cloned, sequenced, expressed and characterizeda-amylase from the clinical isolate of S.aureus resistant to vancomycin (VRSA). The 25kb plasma DNAwas isolated from the clinical isolate of S.aureus and a 600bp PCR amplified product was obtained.Results: The SDS-PAGE analysis of induced and un-induced clone SB3 indicates that a-amylase has aM.W. of 22KD. The ra-amylase was eluted from the gel and enzymes assay was performed. The Km wasfound to be close to Km of native a-amylase identified in the extracellular and in the cytosolic fraction ofS.aureus resistant to vancomycin.Conclusion: The current study clearly indicate that there is clear distinction between the a-amylase ofhuman and different prokaryotes, particularly S.aureus which made this enzyme an ideal target in thedevelopment of drugs against infections of tooth and oral cavities.Keywords: Staphylococcus aureus, Alpha-amylase, Cloning, Vancomycin.

Cloning and characterization of alpha-amylase from a clinical isolate of staphylococcus aureus resistant to vancomycin