Bacteriophage endolysin Lyt μ1/6: characterization of the C-terminal binding domain

Authors

  • Lenka Tišáková,

    Corresponding author
    1. Department of Genomics and Biotechnology, Laboratory of Prokaryotic Biology, Institute of Molecular Biology Slovak Academy of Sciences (IMB SAS), Bratislava, Slovakia
    • Correspondence: Lenka Tišáková, Institute of Molecular Biology Slovak Academy of Sciences (IMB SAS), Dúbravská cesta 21, SK-84551 Bratislava, Slovakia. Tel.: +421 2 59307432; fax: +421 2 59307416; e-mail: lenka.tisakova@savba.sk

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  • Barbora Vidová,

    1. Department of Genomics and Biotechnology, Laboratory of Prokaryotic Biology, Institute of Molecular Biology Slovak Academy of Sciences (IMB SAS), Bratislava, Slovakia
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  • Jarmila Farkašovská,

    1. Department of Genomics and Biotechnology, Laboratory of Prokaryotic Biology, Institute of Molecular Biology Slovak Academy of Sciences (IMB SAS), Bratislava, Slovakia
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  • Andrej Godány

    1. Department of Genomics and Biotechnology, Laboratory of Prokaryotic Biology, Institute of Molecular Biology Slovak Academy of Sciences (IMB SAS), Bratislava, Slovakia
    2. Faculty of Natural Sciences, Department of Biotechnology, University of Ss. Cyril and Methodius in Trnava, Trnava, Slovakia
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Abstract

The gene product of orf50 from actinophage μ1/6 of Streptomyces aureofaciens is a putative endolysin, Lyt μ1/6. It has a two-domain modular structure, consisting of an N-terminal catalytic and a C-terminal cell wall binding domain (CBD). Comparative analysis of Streptomyces phage endolysins revealed that they all have a modular structure and contain functional C-terminal domains with conserved amino acids, probably associated with their binding function. A blast analysis of Lyt μ1/6 in conjunction with secondary and tertiary structure prediction disclosed the presence of a PG_binding_1 domain within the CBD. The sequence of the C-terminal domain of lyt μ1/6 and truncated forms of it were cloned and expressed in Escherichia coli. The ability of these CBD variants fused to GFP to bind to the surface of S. aureofaciens NMU was shown by specific binding assays.

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