The spontaneous host-range mutants 812F1 and K1/420 are derived from polyvalent phage 812 that is almost identical to phage K, belonging to family Myoviridae and genus Kayvirus. Phage K1/420 is used for the phage therapy of staphylococcal infections. Endolysin of these mutants designated LysF1, consisting of an N-terminal cysteine-histidine-dependent aminohydrolase/peptidase (CHAP) domain and C-terminal SH3b cell wall-binding domain, has deleted middle amidase domain compared to wild-type endolysin. In this work, LysF1 and both its domains were prepared as recombinant proteins and their function was analyzed. LysF1 had an antimicrobial effect on 31 Staphylococcus species of the 43 tested. SH3b domain influenced antimicrobial activity of LysF1, since the lytic activity of the truncated variant containing the CHAP domain alone was decreased. The results of a co-sedimentation assay of SH3b domain showed that it was able to bind to three types of purified staphylococcal peptidoglycan 11.2, 11.3, and 11.8 that differ in their peptide bridge, but also to the peptidoglycan type 11.5 of Streptococcus uberis, and this capability was verified in vivo using the fusion protein with GFP and fluorescence microscopy. Using several different approaches, including NMR, we have not confirmed the previously proposed interaction of the SH3b domain with the pentaglycine bridge in the bacterial cell wall. The new naturally raised deletion mutant endolysin LysF1 is smaller than LysK, has a broad lytic spectrum, and therefore is an appropriate enzyme for practical use. The binding spectrum of SH3b domain covering all known staphylococcal peptidoglycan types is a promising feature for creating new chimeolysins by combining it with more effective catalytic domains.

CEITEC Central European Institute of Technology Masaryk University Kamenice 5 Brno 625 00 Czech Republic

Department of Biology Faculty of Natural Sciences University of Ss Cyril and Methodius in Trnava Hlavná 418 919 51 Špačince Slovak Republic

Department of Experimental Biology Faculty of Science Masaryk University Kotlářská 2 Brno 611 37 Czech Republic

National Centre for Biomolecular Research Faculty of Science Masaryk University Kamenice 5 Brno 625 00 Czech Republic

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Nat Rev Microbiol. 2010 May;8(5):317-27 PubMed

Future Microbiol. 2012 Oct;7(10):1147-71 PubMed

Adv Virus Res. 2012;83:143-216 PubMed

Appl Environ Microbiol. 2009 Feb;75(3):872-4 PubMed

Curr Microbiol. 2011 Dec;63(6):538-42 PubMed

Front Microbiol. 2015 Nov 25;6:1315 PubMed

Virology. 1998 Jul 5;246(2):241-52 PubMed

J Biomol NMR. 1995 Nov;6(3):277-93 PubMed

Nucleic Acids Res. 2015 Jan;43(Database issue):D213-21 PubMed

Genome Announc. 2014 Jan 23;2(1):null PubMed

Biosens Bioelectron. 2016 Mar 15;77:366-71 PubMed

J Biotechnol. 2009 Jan 1;139(1):118-23 PubMed

FEMS Microbiol Lett. 2013 May;342(1):30-6 PubMed

Anal Biochem. 2006 Oct 15;357(2):289-98 PubMed

Appl Biochem Biotechnol. 2016 Oct;180(3):544-557 PubMed

Res Vet Sci. 2017 Apr;111:113-119 PubMed

Gene. 2009 Aug 15;443(1-2):32-41 PubMed

J Bacteriol. 2004 May;186(9):2862-71 PubMed

PLoS Pathog. 2014 May 15;10(5):e1004109 PubMed

Appl Environ Microbiol. 2005 Apr;71(4):1836-42 PubMed

Folia Microbiol (Praha). 2007;52(4):331-8 PubMed

FEBS J. 2014 Sep;281(18):4112-22 PubMed

FEMS Microbiol Lett. 2015 Jan;362(1):1-8 PubMed

Appl Environ Microbiol. 2006 Apr;72(4):2988-96 PubMed

J Bacteriol. 2006 Apr;188(7):2463-72 PubMed

Int J Microbiol. 2013;2013:625341 PubMed

Proc Natl Acad Sci U S A. 2016 Aug 16;113(33):9351-6 PubMed

J Appl Microbiol. 2016 Sep;121(3):618-33 PubMed

PLoS One. 2011;6(9):e24418 PubMed

Chembiochem. 2012 May 7;13(7):959-63 PubMed

Microb Biotechnol. 2011 Sep;4(5):643-50 PubMed

Appl Environ Microbiol. 2012 Apr;78(7):2241-8 PubMed

Front Microbiol. 2016 Apr 08;7:474 PubMed

Virol J. 2014 Jul 26;11:133 PubMed

Sci Rep. 2017 Jan 24;7:41259 PubMed

FEMS Microbiol Lett. 2014 Jan;350(2):199-208 PubMed

FEMS Microbiol Lett. 2009 May;294(1):52-60 PubMed

J Biomol NMR. 2009 Aug;44(4):213-23 PubMed

J Appl Microbiol. 2011 Oct;111(4):1025-35 PubMed

Nucleic Acids Res. 1998 Feb 15;26(4):1107-15 PubMed

Can J Microbiol. 2000 Nov;46(11):1066-76 PubMed

J Biol Chem. 2006 Jan 6;281(1):549-58 PubMed

Arch Virol. 2016 Oct;161(10 ):2921-49 PubMed

Proteomics. 2007 Jan;7(1):64-72 PubMed

J Bacteriol. 2005 Oct;187(20):7161-4 PubMed

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