摘要
Klebsiella pneumoniae is one of the major pathogens causing global multidrug-resistant infections.Therefore,strategies for preventing and controlling the infections are urgently needed.Phage depolymerase,often found in the tail fiber protein or the tail spike protein,is reported to have antibiofilm activity.In this study,phage P560isolated from sewage showed specific for capsule locus type KL47 K.pneumoniae,and the enlarged haloes around plaques indicated that P560 encoded a depolymerase.The capsule depolymerase,ORF43,named P560dep,derived from phage P560 was expressed,purified,characterized and evaluated for enzymatic activity as well as specificity.We reported that the capsule depolymerase P560dep,can digest the capsule polysaccharides on the surface of KL47 type K.pneumoniae,and the depolymerization spectrum of P560dep matched to the host range of phage P560,KL47 K.pneumoniae.Crystal violet staining assay showed that P560dep was able to significantly inhibit biofilm formation.Further,a single dose(50μg/mouse)of depolymerase intraperitoneal injection protected 90%–100%of mice from lethal challenge before or after infection by KL47 carbapenem-resistant K.pneumoniae.And pathological changes were alleviated in lung and liver of mice infected by KL47 type K.pneumoniae.It is demonstrated that depolymerase P560dep as an attractive antivirulence agent represents a promising tool for antimicrobial therapy.
基金
the National Natural Science Foundation of China(U1803109)
the National Key Research and Development Program of China(2018YFC1602500)
the National Natural Science Foundation of China(81572032)
Major scientific and technological innovation projects in Shandong Province(2019JZZY010719)
National Key Research and Development Program(2019YFA0904003)
Six Talent Peaks Project in Jiangsu Province(2016-WSN-112)
Key research and development project of Jiangsu provincial science and Technology Department(BE2017654)
Gusu key health talent of Suzhou,Jiangsu youth medical talents program(QN-867)
the Science and Technology Program of Suzhou(SZS201715)。
