In Vitro Synthesis of Bioactive Glycovariants of Enterocin 96, an Antimicrobial Peptide from Enterococcus faecalis

  • Rupa Nagar
  • Alka RaoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1954)


Antimicrobial peptides (AMPs) are novel agents for therapeutic application for their inherent broad spectrum of activities against bacteria, fungi, and viruses, as well as anti-inflammatory, anticancerous, and immunomodulatory activities. This chapter presents an enzymatic method to generate glycovariants of one such antimicrobial peptide, namely enterocin 96, using a bacterial protein O- and S-glycosyltransferase, in vitro.

Key words

In vitro glycosyltransferase assay Mass spectrometry High performance liquid chromatography (HPLC) Chymotrypsin digestion Exoglycosidase treatment/Deglycosylation assay Glycodiversification 


  1. 1.
    Schäffer C, Messner P (2017) Emerging facets of prokaryotic glycosylation. FEMS Microbiol Rev 41:49–91CrossRefGoogle Scholar
  2. 2.
    Maky MA, Ishibashi N, Zendo T et al (2015) Enterocin F4-9, a novel O-linked glycosylated bacteriocin. Appl Environ Microbiol 81:4819–4826CrossRefGoogle Scholar
  3. 3.
    Nagar R, Rao A (2017) An iterative glycosyltransferase EntS catalyzes transfer and extension of O- and S-linked monosaccharide in enterocin 96. Glycobiology 27:766–776CrossRefGoogle Scholar
  4. 4.
    Wang H, Oman TJ, Zhang R et al (2014) The glycosyltransferase involved in thurandacin biosynthesis catalyzes both O- and S-glycosylation. J Am Chem Soc 136:84–87CrossRefGoogle Scholar
  5. 5.
    Stepper J, Shastri S, Loo TS et al (2011) Cysteine S-glycosylation, a new post-translational modification found in glycopeptide bacteriocins. FEBS Lett 585:645–650CrossRefGoogle Scholar
  6. 6.
    Oman TJ, Boettcher JM, Wang H et al (2011) Sublancin is not a lantibiotic but an S-linked glycopeptide. Nat Chem Biol 7:78–80CrossRefGoogle Scholar
  7. 7.
    Izquierdo EC, Wagner C et al (2009) Enterocin 96, a novel class II bacteriocin produced by enterococcus faecalis WHE 96, isolated from Munster cheese. Appl Environ Microbiol 75:4273–4276CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.CSIR-Institute of Microbial Technology (CSIR-IMTECH)ChandigarhIndia

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