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Purification and Characterization of a New Halocin HA4 from Haloferax larsenii HA4 Isolated from a Salt Lake

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Abstract

Halocins are antimicrobial peptides secreted by different members of haloarchaea. Halocin HA4 was purified from Haloferax larsenii HA4 using combination of ultrafiltration and chromatographic techniques. It was found to be ~ 14 kDa with unique N-terminal sequence, H2N-AEEEIFXPDX, which did not show homology with the known sequence suggesting a new/novel compound. It was found to be heat resistant up to 100 °C, stable at pH 2.0–10.0, and retained complete activity in the presence of different organic compounds such as methanol, ethanol, acetone, isopropanol, ethyl acetate, Tween 80, acetonitrile, SDS, Triton X-100, and urea. However, complete activity was reduced after the treatment with trypsin, papain, and proteinase K suggesting proteinaceous nature of the compound. The cytocidal nature of halocin HA4 was evidenced with complete loss of viable count of indicator strain, H. larsenii HA10. The change in FTIR spectrum of halocin-treated cells suggested halocin HA4 interacts with cell membrane and nucleic acids of the target cells. Thus, we report a new halocin inhibitory to related strains and may be applied in the preservation of salted foods and leather hides in the respective industries.

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Materials described in the manuscript, including all relevant raw data, will be freely available to any researcher wishing to use them for non-commercial purposes, without breaching participant confidentiality.

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Funding

This work was financially supported by the Council of Scientific and Industrial Research, New Delhi (38(1220)/09/EMR-II), and Department of Biotechnology (DBT), Ministry of Science and Technology, New Delhi (BT/PR8306/PID/6/738/2013).

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  1. Department of Genetics, Maharshi Dayanand University, Rohtak, 124001, Haryana, India

    Ramanjeet Kaur & Santosh Kumar Tiwari

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  1. Ramanjeet Kaur
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  2. Santosh Kumar Tiwari
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Correspondence to Santosh Kumar Tiwari.

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Kaur, R., Tiwari, S.K. Purification and Characterization of a New Halocin HA4 from Haloferax larsenii HA4 Isolated from a Salt Lake. Probiotics & Antimicro. Prot. 13, 1458–1466 (2021). https://doi.org/10.1007/s12602-021-09823-2

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