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Biosynthesis of silver nanoparticles using Bacillus subtilis EWP-46 cell-free extract and evaluation of its antibacterial activity

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Abstract

This study highlights the ability of nitrate-reducing Bacillus subtilis EWP-46 cell-free extract used for preparation of silver nanoparticles (AgNPs) by reduction of silver ions into nano silver. The production of AgNPs was optimized with several parameters such as hydrogen ion concentration, temperature, silver ion (Ag+ ion) and time. The maximum AgNPs production was achieved at pH 10.0, temperature 60 °C, 1.0 mM Ag+ ion and 720 min. The UV–Vis spectrum showed surface plasmon resonance peak at 420 nm, energy-dispersive X-ray spectroscopy (SEM–EDX) spectra showed the presence of element silver in pure form. Atomic force microscopy (AFM) and transmission electron microscopy images illustrated the nanoparticle size, shape, and average particle size ranging from 10 to 20 nm. Fourier transform infrared spectroscopy provided the evidence for the presence of biomolecules responsible for the reduction of silver ion, and X-ray diffraction analysis confirmed that the obtained nanoparticles were in crystalline form. SDS-PAGE was performed to identify the proteins and its molecular mass in the purified nitrate reductase from the cell-free extract. In addition, the minimum inhibitory concentration and minimum bactericidal concentration of AgNPs were investigated against gram-negative (Pseudomonas fluorescens) and gram-positive (Staphylococcus aureus) bacteria.

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Acknowledgments

This research was supported by the Korean National Research Foundation (Korean Ministry of Education, Science and Technology, Award NRF-2011-35B-D00020). The preparation of the manuscript was supported by research funds from the Chonbuk National University in 2013. This work was also partially supported by Korea Ministry of Environment as “The GAIA Project (No. 2012000550021)”.

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Authors and Affiliations

  1. Division of Biotechnology, Advanced Institute of Environment and Bioscience, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan, Jeonbuk, 570-752, South Korea

    Palanivel Velmurugan, Min Cho & Byung-Taek Oh

  2. Department of Environmental Sciences, Bharathiar University, Coimbatore, Tamil Nadu, 641-046, India

    Mahudunan Iydroose

  3. Department of Microbiology, Sadakathullah Appa College, Tirunelveli, Tamil Nadu, 627002, India

    Mohmed Hanifa Abdul Kader Mohideen

  4. Department of Zoology, Rani Anna Govt. Arts and Science College, Tirunelveli, Tamil Nadu, 627008, India

    Thankiah Selva Mohan

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  1. Palanivel Velmurugan
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  2. Mahudunan Iydroose
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  3. Mohmed Hanifa Abdul Kader Mohideen
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  4. Thankiah Selva Mohan
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  5. Min Cho
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  6. Byung-Taek Oh
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Correspondence to Byung-Taek Oh.

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Velmurugan, P., Iydroose, M., Mohideen, M.H.A.K. et al. Biosynthesis of silver nanoparticles using Bacillus subtilis EWP-46 cell-free extract and evaluation of its antibacterial activity. Bioprocess Biosyst Eng 37, 1527–1534 (2014). https://doi.org/10.1007/s00449-014-1124-6

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  • DOI: https://doi.org/10.1007/s00449-014-1124-6

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