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Indexed/Abstracted in: EMBASE, Science Citation Index Expanded (SciSearch), Scopus
Impact Factor 0,246
Online ISSN 1827-160X
Younes GHASEMI 1, 3, 4 , Nasim HAJIGHAHRAMANI 1, 2, 3, Fatemeh DABBAGH 1, 3, Mohammad B. GHOSHOON 1, 3, Elham YARAHMADI 2, 3, Mohammad A. MOBASHER 3, 5, 6, Nima MONTAZERI-NAJAFABADY 1, 3
1 Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran; 2 Student Research Committee, International Branch, Shiraz University of Medical Sciences, Shiraz, Iran; 3 Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; 4 Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran; 5 Noncommunicable Diseases Research Center, Fasa University of Medical Sciences, Fasa, Iran; 6 Department of Medical Biotechnology, School of Medicine, Fasa University of Medical Sciences, Fasa, Iran
BACKGROUND: The multifunctional enzyme α-enolase belongs to a family of cytoplasmic and glycolytic enzymes, which is localized in the cytoplasm and at the surface of many eukaryotic and prokaryotic cells. α-enolase on the surface of Staphylococcus aureus (S. aureus) is a receptor with high affinity for laminin, the most abundant extracellular matrix component. Laminin-binding ability plays a considerable role in the pathogenesis of this microorganism. S. aureus is an opportunistic pathogen that causes major nosocomial infections and variety of diseases in human beings. This organism has a strong tendency to develop antibiotic resistance. Its property to spread of antibiotic resistance has intensified the need of novel antistaphylococcal techniques. α-enolase, as an important surface antigen, is a potential vaccine or immunotherapeutic candidate.
METHODS: For cloning and expression studies, α-enolase gene was amplified by PCR using the specific primers. Then it was inserted into the pET-15b vector and transformed in to Escherichia coli DE3. Gene expression was induced at 30° C using IPTG and the recombinant enzyme purification carried out by Ni-NTA spin columns. Protein was shown with SDS-PAGE analysis and then, different induction conditions for optimization of the recombinant protein expression were used.
RESULTS: The highest protein expression was observed when the α-enolase production was induced using the mixture of two different inducers at the same time.
CONCLUSION: We cloned, expressed, and purified α-enolase from S. aureus which could be a significant step for applying the recombinant enzyme in biotechnological processes.