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Purification and structural characterization of a novel antibacterial peptide from Bellamya bengalensis: activity against ampicillin and chloramphenicol resistant Staphylococcus epidermidis
Authors:Gauri Samiran S  Mandal Santi M  Pati Bikas R  Dey Satyahari
Affiliation:a Central Research Facility, Indian Institute of Technology Kharagpur, Kharagpur 721302, WB, India
b Department of Microbiology, Vidyasagar University, Midnapore 721102, WB, India
c Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302, WB, India
Abstract:
Increasing tendency of clinical bacterial strains resistant to conventional antibiotics has being a great challenge to the public's health. Antimicrobial peptides, a new class of antibiotics is known to have the activity against a wide range of bacteria resistant to conventional antibiotics. An antimicrobial peptide of 1676 Da was purified from Bellamya bengalensis, a fresh water snail, using ultrafiltration and reversed phase liquid chromatography. The effect of this peptide on Staphylococcus epidermidis resistant to ampicillin and chloramphenicol was investigated; the MIC and MBC values were 8 μg/ml and 16 μg/ml, respectively. Complete sequence of the peptide was determined by tandem mass spectrometry (MS/MS). Further, peptide net charge, hydrophobicity and molecular modeling were evaluated in silico for better understanding the probable mechanisms of action. The peptide showed the specificity to bacterial membranes. Hence, this reported peptide revealed a promising candidate to contribute in the development of therapeutic agent for Staphylococcal infections.
Keywords:AMP, antimicrobial peptides   Bb-AMP4, Bellamya bengalensis-antimicrobial peptide 4   FACS, fluorescence activated cell sorter   HPLC, high performance liquid chromatography   LC-MS/MS, liquid chromatography-tandem mass spectrometry   MALDI ToF MS, matrix assisted laser desorption ionization time of flight mass spectrometry   MBC, minimum bactericidal concentration   MHA, Mueller-Hinton agar   MHB, Mueller-Hinton broth   MIC, minimum inhibitory concentration   PES, polyethersulfone   PI, propidium iodide   TFA, trifluoroacetic acid
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