PMID:26085488

Citation	Ugorcakova, J, Medzova, L, Solteszova, B and Bukovska, G (2015) Characterization of a phiBP endolysin encoded by the Paenibacillus polymyxa CCM 7400 phage. FEMS Microbiol. Lett. 362:fnv098
Abstract	Endolysin (gp1.2) from the Paenibacillus polymyxa CCM 7400 temperate phage phiBP has a modular structure consisting of an N-terminal region with a catalytic glycosyl hydrolase 25 domain and a C-terminal cell wall-binding domain. The entire gene of this endolysin and fragments containing its catalytic and binding domains separately were cloned into expression vectors and the corresponding recombinant proteins were expressed in Escherichia coli and purified by affinity chromatography. The lytic activities of endolysin and its catalytic domain were tested on cell wall substrates from paenibacilli, bacilli, corynebacteria and E. coli. The presence of a cell wall-binding domain was found to be essential, as the phiBP endolysin was fully active only as a full-length protein. The binding ability of the cell wall-binding domain alone and in fusion with green fluorescent protein was demonstrated by specific binding assays to the cell surface of P. polymyxa CCM 7400 and to those of other Paenibacillus strains. Thus the ability of phiBP endolysin to hydrolyze the paenibacilli cell wall was confirmed.
Links	PubMed Online version:10.1093/femsle/fnv098
Keywords	Actinomycetales/metabolism; Amino Acid Sequence; Bacillus/metabolism; Bacteriolysis; Bacteriophages/enzymology; Bacteriophages/genetics; Binding Sites; Catalytic Domain; Cell Wall/metabolism; Computational Biology; Endopeptidases/chemistry; Endopeptidases/genetics; Escherichia coli/genetics; Escherichia coli/metabolism; Green Fluorescent Proteins/genetics; Microscopy, Fluorescence; Paenibacillus/growth & development; Paenibacillus/metabolism; Paenibacillus/ultrastructure; Paenibacillus/virology; Protein Binding; Protein Structure, Tertiary; Recombinant Proteins/chemistry; Sequence Alignment
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
9VIRU:D0R7I3		GO:0019835: cytolysis	ECO:0000314:		P		Figure 4 shows the lytic activity against different hosts. It shows that the protein has specificity against P. polymyxa over other Paenibacillus bacteria.	complete CACAO 12876
9VIRU:D0R7H8		GO:0019835: cytolysis	ECO:0000314:		P		Figure 3 reveals that the protein activity results in clear zones of the bacterial lawn.	complete CACAO 12870

References

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