Home > Riviste > Minerva Biotecnologica > Fascicoli precedenti > Minerva Biotecnologica 2016 March;28(1) > Minerva Biotecnologica 2016 March;28(1):33-8

ULTIMO FASCICOLO
 

ARTICLE TOOLS

Estratti

MINERVA BIOTECNOLOGICA

Rivista di Biologia Molecolare e Biotecnologie


Indexed/Abstracted in: EMBASE, Science Citation Index Expanded (SciSearch), Scopus
Impact Factor 0,25


eTOC

 

ORIGINAL ARTICLES  FREEfree


Minerva Biotecnologica 2016 March;28(1):33-8

Copyright © 2016 EDIZIONI MINERVA MEDICA

lingua: Inglese

Cloning, expression, purification and expression condition optimization of α-enolase from Staphylococcus aureus in Escherichia coli

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


FULL TEXT  


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.

inizio pagina

Publication History

Per citare questo articolo

Corresponding author e-mail