TY - JOUR ID - 5385 TI - LytU-SH3b fusion protein as a novel and efficient enzybiotic against methicillin-resistant Staphylococcus aureus JO - Molecular Biology Research Communications JA - MBRC LA - en SN - 2322-181X AU - Taheri-Anganeh, Mortaza AU - Khatami, Seyyed Hossein AU - Jamali, Zeinab AU - Movahedpour, Ahmad AU - Ghasemi, Younes AU - Savardashtaki, Amir AU - Mostafavi-Pour, Zohreh AD - Department of Medical Biotechnology, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran AD - Recombinant Protein Laboratory, Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran AD - Cardiovascular Research Center, Shiraz University of Medical Sciences, Shiraz, Iran AD - Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran Y1 - 2019 PY - 2019 VL - 8 IS - 4 SP - 151 EP - 158 KW - Staphylococcus aureus KW - Autolysin KW - LytU KW - SH3b KW - Enzybiotics DO - 10.22099/mbrc.2019.34446.1430 N2 - 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. UR - https://mbrc.shirazu.ac.ir/article_5385.html L1 - https://mbrc.shirazu.ac.ir/article_5385_d0e305f52d0982626f4aa57dbee6bc8e.pdf ER -