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Viruses 2018, 10(5), 276;

Discovery and Biochemical Characterization of PlyP56, PlyN74, and PlyTB40—Bacillus Specific Endolysins

Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA
Department of Veterinary Medicine, University of Maryland, College Park, MD 20742, USA
School of Systems Biology and National Center for Biodefense & Infectious Diseases, Biomedical Research Laboratory, George Mason University, Manassas, VA 22030, USA
Department of Integrated Science & Technology, James Madison University, Harrisonburg, VA 22807, USA
Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, VA 23220, USA
Department of Medicinal Chemistry and Institute for Structural Biology, Drug Discovery and Development, School of Pharmacy, Virginia Commonwealth University, Richmond, VA 23220, USA
Current address: Department of Biochemistry and Biophysics, Texas A & M University, College Station, TX 77843, USA.
Author to whom correspondence should be addressed.
Received: 30 April 2018 / Revised: 15 May 2018 / Accepted: 17 May 2018 / Published: 21 May 2018
(This article belongs to the Special Issue Phage Lytic Enzymes and Their Applications)
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Three Bacillus bacteriophage-derived endolysins, designated PlyP56, PlyN74, and PlyTB40, were identified, cloned, purified, and characterized for their antimicrobial properties. Sequence alignment reveals these endolysins have an N-terminal enzymatically active domain (EAD) linked to a C-terminal cell wall binding domain (CBD). PlyP56 has a Peptidase_M15_4/VanY superfamily EAD with a conserved metal binding motif and displays biological dependence on divalent ions for activity. In contrast, PlyN74 and PlyTB40 have T7 lysozyme-type Amidase_2 and carboxypeptidase T-type Amidase_3 EADs, respectively, which are members of the MurNAc-LAA superfamily, but are not homologs and thus do not have a shared protein fold. All three endolysins contain similar SH3-family CBDs. Although minor host range differences were noted, all three endolysins show relatively broad antimicrobial activity against members of the Bacillus cereus sensu lato group with the highest lytic activity against B. cereus ATCC 4342. Characterization studies determined the optimal lytic activity for these enzymes was at physiological pH (pH 7.0–8.0), over a broad temperature range (4–55 °C), and at low concentrations of NaCl (<50 mM). Direct comparison of lytic activity shows the PlyP56 enzyme to be twice as effective at lysing the cell wall peptidoglycan as PlyN74 or PlyTB40, suggesting PlyP56 is a good candidate for further antimicrobial development as well as bioengineering studies. View Full-Text
Keywords: endolysin; bacteriophage; Bacillus cereus sensu lato; peptidoglycan hydrolase endolysin; bacteriophage; Bacillus cereus sensu lato; peptidoglycan hydrolase

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Etobayeva, I.; Linden, S.B.; Alem, F.; Harb, L.; Rizkalla, L.; Mosier, P.D.; Johnson, A.A.; Temple, L.; Hakami, R.M.; Nelson, D.C. Discovery and Biochemical Characterization of PlyP56, PlyN74, and PlyTB40—Bacillus Specific Endolysins. Viruses 2018, 10, 276.

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