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Identification and characterization of antimicrobial peptide produced by indigenously isolated Bacillus paralicheniformis UBBLi30 strain

Abstract

The antimicrobial compound produced by Bacillus paralicheniformis UBBLi30 showed UV spectra absorption at 208 nm. Fourier transform infrared (FTIR) revealed characteristic bands for aliphatic chain related to hydrophobic amino acids (l-isoleucine/l-leucine) (3068, 2965 and 2871 cm−1) and peptide bonds (1538, 1667 and 3312 cm−1). The proton nuclear magnetic resonance (1H NMR) showed signals for aromatic amino acid (6.5–9.5 ppm) and alkyl amines (3–4 ppm). The results of carbon (13C) NMR showed signals for aromatic, nitro and amide compounds. Besides this, the mass fragments (1422.576 [M+H]+, 711.912 [M+2H]2+ and 475.174 [M+3H]3+ m/z) observed in electrospray ionization mass spectrometry (ESI-MS) were coordinated well to the fragments of polypeptide antibiotic bacitracin. The presence of bacA gene further confirmed the production of bacitracin. Bacitracin inhibited the growth of a range of Gram-positive bacteria such as Micrococcus luteus, methicillin-resistant Staphylococcus aureus (MRSA), S. aureus, Streptococcus pyogenes and Propionibacterium acnes, and biofilm formation of M. luteus and MRSA. Moreover, this polypeptide reduced the zeta potential of M. luteus and MRSA, indicating the electrostatic sorption on bacterial surface and concentration-dependent cell membrane damages. Besides this, polypeptide showed stability in the presence of proteases (proteinase K, trypsin and pepsin), pH (1, 3, 5, 7, 9 and 11) and temperature up to 100 °C. B. paralicheniformis UBBLi30 therefore has the potential to be utilized as a bio-preservative to control the growth of spoilage and pathogenic bacteria.

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Author information

JJA, LSG and RSM contributed to the study conception. JJA designed the experiments. Material preparation, data collection and analysis were performed by JJA and MSK. The first draft of the manuscript was written by JJA and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Correspondence to J. J. Ahire.

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Conflict of interest

The authors disclose that financial support for the research described in the manuscript was provided by Unique Biotech Limited, Hyderabad, India. Dr. Ratna Sudha Madempudi is the Managing Director of Unique Biotech Limited. This does not alter our adherence to journal policies on sharing data and materials.

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The research conducted for this article did not involve studies on humans or animals.

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Ahire, J.J., Kashikar, M.S., Lakshmi, S.G. et al. Identification and characterization of antimicrobial peptide produced by indigenously isolated Bacillus paralicheniformis UBBLi30 strain. 3 Biotech 10, 112 (2020). https://doi.org/10.1007/s13205-020-2109-6

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Keywords

  • Bacillus paralicheniformis UBBLi30
  • Bacitracin
  • Zeta potential
  • Biofilm
  • MRSA