Characterization of PlyB221 and PlyP32, Two Novel Endolysins Encoded by Phages Preying on theBacillus cereusGroup

被引:12
作者
Leprince, Audrey [1 ]
Nuytten, Manon [1 ]
Gillis, Annika [1 ,2 ,3 ]
Mahillon, Jacques [1 ]
机构
[1] UCLouvain, Earth & Life Inst, Lab Food & Environm Microbiol, B-1348 Louvain La Neuve, Belgium
[2] Imperial Coll London, Sect Mol Microbiol, London WC2N 5DU, England
[3] Imperial Coll London, MRC Ctr Mol Bacteriol & Infect, London WC2N 5DU, England
来源
VIRUSES-BASEL | 2020年 / 12卷 / 09期
关键词
bacteriophage; endolysin; Bacillus cereus; foodborne pathogen; antimicrobial agent; detection tool; BACILLUS-CEREUS; SP NOV; PSYCHROTOLERANT; BINDING; AGGREGATION; SUBTILIS; SEQUENCE; SEARCH; RANGE;
D O I
10.3390/v12091052
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Endolysins are phage-encoded enzymes implicated in the breaching of the bacterial cell wall at the end of the viral cycle. This study focuses on the endolysins of Deep-Blue (PlyB221) and Deep-Purple (PlyP32), two phages preying on theBacillus cereusgroup. Both enzymes exhibit a typical modular organization with an enzymatically active domain (EAD) located in the N-terminal and a cell wall binding domain (CBD) in the C-terminal part of the protein. In silico analysis indicated that the EAD domains of PlyB221 and PlyP32 are endowed with peptidase and muramidase activities, respectively, whereas in both proteins SH3 domains are involved in the CBD. To evaluate their antimicrobial properties and binding specificity, both endolysins were expressed and purified. PlyB221 and PlyP32 efficiently recognized and lysed all the tested strains from theB. cereusgroup. Biochemical characterization showed that PlyB221 activity was stable under a wide range of pHs (5-9), NaCl concentrations (up to 200 mM), and temperature treatments (up to 50 degrees C). Although PlyP32 activity was less stable than that of PlyB221, the endolysin displayed high activity at pH 6-7, NaCl concentration up to 100 mM and the temperature treatment up to 45 degrees C. Overall, PlyB221 and PlyP32 display suitable characteristics for the development of biocontrol and detection tools.
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页数:16
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