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Phage vB_KpnM_JYSS3 encodes a novel polysaccharide depolymerase that exhibits specific activity against K2-type

Shi-Yan Jiao1, Xia Wang1, Jie Wang1

  • 1College of Lab Medicine, Hebei North University, Zhangjiakou, Hebei Province, 075000, People's Republic of China.

BMC Microbiology
|October 22, 2025
PubMed
Abstract

Insights

A newly discovered phage, vB_KpnM_JYSS3, effectively targets and lyses carbapenem-resistant Klebsiella pneumoniae (CRKP). Its tail protein, Dep17, degrades bacterial capsules and enhances antibiotic efficacy against CRKP biofilms.

Area of Science:

  • Microbiology
  • Bacteriology
  • Virology

Background:

  • Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a significant nosocomial pathogen causing severe infections.
  • The rise of CRKP necessitates novel therapeutic strategies due to increasing antimicrobial resistance.

Purpose of the Study:

  • To isolate and characterize a novel bacteriophage targeting K2-type CRKP.
  • To investigate the antibiofilm activity of the phage and its potential therapeutic applications.
  • To identify and analyze specific phage proteins involved in CRKP lysis and biofilm disruption.

Main Methods:

  • Isolation of bacteriophages from hospital sewage using double-layer agar plates with CRKP.
  • Comprehensive characterization of the isolated phage vB_KpnM_JYSS3, including optimal multiplicity of infection, lytic spectrum, stability, and growth kinetics.
  • Genome sequencing of the phage and prediction of coding sequences.
  • Expression, purification, and functional analysis of the phage tail protein (ORF17), named Dep17.
  • Assessment of antibiofilm activity using 96-well microtitration plates and determination of Dep17's effect on capsular polysaccharide (CPS).

Main Results:

  • Isolation and identification of a novel lytic phage, vB_KpnM_JYSS3, specific for K2-type CRKP.
  • Phage vB_KpnM_JYSS3 demonstrated significant antibiofilm activity, preventing biofilm formation and degrading mature biofilms.
  • Genome sequencing revealed vB_KpnM_JYSS3 as a nontemperate phage with a genome size of 47,244 bp, containing 71 predicted coding sequences.
  • The tail protein, Dep17, was confirmed to possess depolymerase activity, degrading the CPS of K2-type CRKP.
  • Dep17 enhanced the efficacy of polymyxin B against CRKP biofilms.

Conclusions:

  • Phage vB_KpnM_JYSS3 represents a promising lytic agent against K2-type CRKP.
  • The novel depolymerase, Dep17, shows potential for disrupting CRKP biofilms and enhancing antibiotic treatment.
  • Combined phage and depolymerase therapy could offer a novel strategy for combating drug-resistant CRKP infections.
Keywords:
BacteriophageBiofilmCarbapenem-resistant Klebsiella pneumoniae (CRKP)Depolymerase

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