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Anti-methicillin Resistant Staphylococcus aureus Compound Isolation from Halophilic Bacillus amyloliquefaciens MHB1 and Determination of Its Mode of Action Using Electron Microscope and Flow Cytometry Analysis

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Abstract

The aim of this study was to purify, characterize and evaluate the antibacterial activity of bioactive compound against methicillin-resistant Staphylococcus aureus (MRSA). The anti-MRSA compound was produced by a halophilic bacterial strain designated as MHB1. The MHB1 strain exhibited 99 % similarity to Bacillus amyloliquefaciens based on 16S rRNA gene analysis. The culture conditions of Bacillus amyloliquefaciens MHB1 were optimized using nutritional and environmental parameters for enhanced anti-MRSA compound production. The pure bioactive compound was isolated using silica gel column chromatography and Semi-preparative High-performance liquid chromatography (Semi-preparative HPLC). The Thin layer chromatography, Fourier transform infrared spectroscopy and proton NMR (1H NMR) analysis indicated the phenolic nature of the compound. The molecular mass of the purified compound was 507 Da as revealed by Liquid chromatography-mass spectrometry (LC–MS) analysis. The compound inhibited the growth of MRSA with minimum inhibitory concentration (MIC) of 62.5 µg mL−1. MRSA bacteria exposed to 4× MIC of the compound and the cell viability was determined using flow cytometric analysis. Scanning electron microscope and Transmission electron microscope analysis was used to determine the ultrastructural changes in bacteria. This is the first report on isolation of anti-MRSA compound from halophilic B. amyloliquefaciens MHB1 and could act as a promising biocontrol agent.

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Acknowledgments

The authors wish to thank the management of SRM University for providing necessary facilities for undertaking this study. The authors would like to thank the Nanotechnology Research Center for SEM analysis, Interdisciplinary Institute of Indian System of Medicine (IIISM) lab for flow cytometry analysis at SRM University and Tamil Nadu Veterinary and Animal Sciences University (TANUVAS), Chennai for TEM analysis.

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  1. Department of Biotechnology, School of Bioengineering, SRM University, SRM Nagar, Kattankulathur, Chennai, 603 203, India

    Venkadapathi Jeyanthi & Palaniyandi Velusamy

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  1. Venkadapathi Jeyanthi
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  2. Palaniyandi Velusamy
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Correspondence to Palaniyandi Velusamy.

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Jeyanthi, V., Velusamy, P. Anti-methicillin Resistant Staphylococcus aureus Compound Isolation from Halophilic Bacillus amyloliquefaciens MHB1 and Determination of Its Mode of Action Using Electron Microscope and Flow Cytometry Analysis. Indian J Microbiol 56, 148–157 (2016). https://doi.org/10.1007/s12088-016-0566-8

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