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LytU-SH3b fusion protein as a novel and efficient enzybiotic against methicillin-resistant Staphylococcus aureus | ||
| Molecular Biology Research Communications | ||
| مقاله 2، دوره 8، شماره 4، 2019، صفحه 151-158 اصل مقاله (345.67 K) | ||
| نوع مقاله: Original article | ||
| شناسه دیجیتال (DOI): 10.22099/mbrc.2019.34446.1430 | ||
| نویسندگان | ||
| Mortaza Taheri-Anganeh1؛ Seyyed Hossein Khatami2؛ Zeinab Jamali3؛ Ahmad Movahedpour1، 4؛ Younes Ghasemi5، 6؛ Amir Savardashtaki1، 6؛ Zohreh Mostafavi-Pour* 2، 7 | ||
| 1Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| 2Recombinant Protein Laboratory, Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| 3Cardiovascular Research Center, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| 4Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| 5Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| 6Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| 7Maternal-Fetal Medicine Research Center, Shiraz University of Medical Sciences, Shiraz, Iran | ||
| چکیده | ||
| Methicillin-resistant Staphylococcus aureus (MRSA) is a challenging infectious agent worldwide. The ever growing antibiotic resistance has made the researchers to look for new anti-staphylococcal agents. Autolysins are staphylococcal enzymes that lyse bacterial cell wall for cell division. Autolysins can be used as novel enzybiotics (enzymes have antibiotic effects) for staphylococcal infections. LytU is a newly explored autolysin. SH3b is a potent cell wall binding domain that can be fused to lytic enzymes to increase their activity. The aim of this study was to design a novel and efficient fusion enzybiotic that could lyse staphylococcal cell wall peptidoglycan by disrupting the bacteria. LytU-SH3b fusion construct was synthesized and LytU was amplified through the construct, using overhang PCR. The fusion and native forms that had his-tag were synthesized by recombinant technology in Escherichia coli BL21 (DE3) strain and purified utilizing Ni-NTA agarose beads. LytU and LytU-SH3b activity and potency were assessed using plate lysis assay, turbidity reduction assay and minimal inhibitory concentration (MIC) tests. All these tests showed that LytU-SH3b has more activity and potency than LytU. LytU-SH3b has MIC 421 fold lesser than LytU. Finally, LytU-SH3b is a novel and efficient recombinant enzybiotic that can lyse MRSA as an alternative to chemical small molecule antibiotics. | ||
| کلیدواژهها | ||
| Staphylococcus aureus؛ Autolysin؛ LytU؛ SH3b؛ Enzybiotics | ||
| مراجع | ||
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