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Purification and Characterization of Enterocin LR/6, a Bacteriocin from Enterococcus faecium LR/6

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Abstract

Enterocin LR/6, a bacteriocin obtained from the culture filtrate of Enterococcus faecium strain LR/6, has been purified to homogeneity using ammonium sulfate precipitation, cation-exchange chromatography, gel-filtration, and checked on reverse-phase high-performance liquid chromatography. It is active at high temperatures (boiling as well as autoclaving) and over a wide range of pH (2.0–8.0). Also, it is sensitive to a number of proteolytic enzymes but is stable in the presence of surfactants and organic solvents. The protein could be stored at least up to 1 year at low temperatures (4 °C and −20 °C) without any loss of activity. The N-terminal sequence of enterocin LR/6 showed no homology with known enterocins or other bacteriocins present in the database, suggesting it to be a novel enterocin. Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed its mass to be ∼6.1 kDa. It showed a bactericidal mode of action against indicator strain, Micrococcus luteus.

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Acknowledgements

This work was financially supported by the Council of Scientific and Industrial Research and the Department of Biotechnology, India. The facilities provided to the Department of Genetics, by University Grant Commission under SAP and by Department of Science and Technology, Government of India under FIST program, is thankfully acknowledged. Author MK was supported by a UGC fellowship.

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  1. Department of Genetics, University of Delhi South Campus, New Delhi, 110021, India

    Manoj Kumar, Santosh Kumar Tiwari & Sheela Srivastava

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  1. Manoj Kumar
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  2. Santosh Kumar Tiwari
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  3. Sheela Srivastava
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Correspondence to Sheela Srivastava.

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Kumar, M., Tiwari, S.K. & Srivastava, S. Purification and Characterization of Enterocin LR/6, a Bacteriocin from Enterococcus faecium LR/6. Appl Biochem Biotechnol 160, 40–49 (2010). https://doi.org/10.1007/s12010-009-8586-z

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  • DOI: https://doi.org/10.1007/s12010-009-8586-z

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