Cloning,expression,and functional analysis of capsule-specific depolymerase targeting carbapenem-resistant Klebsiella pneumoniae
10.19405/j.cnki.issn1000-1492.2025.07.013
- VernacularTitle:耐碳青霉烯类肺炎克雷伯菌荚膜特异性解聚酶克隆表达及其相关功能研究
- Author:
Tao YAN
1
;
Na WANG
;
Qiuyan WANG
;
Chengcheng MA
;
Xuan TENG
;
Kexue YU
;
Honghua GE
;
Zhou LIU
Author Information
1. 安徽医科大学第二附属医院检验科,合肥 230601;安徽省胸科医院临床检验中心,合肥 230022
- Publication Type:Journal Article
- Keywords:
depolymerase;
phage;
carbapenem-resistant Klebsiella pneumoniae;
capsule;
biofilm;
recombinant protein
- From:
Acta Universitatis Medicinalis Anhui
2025;60(7):1251-1257
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the K64 capsule depolymerase recombinant protein,Dep44,and investigate its potential application against carbapenem-resistant Klebsiella pneumoniae(CRKP)infections.Methods The de-polymerase-encoding phage vB_Kpn_HF1013(GenBank:PP803128)was isolated and genomically analyzed to screen for candidate depolymerases.The recombinant protein Dep44 was constructed and functionally verified for depolymerase activity.Dep44 sensitive range was validated and Dep44 antimicrobial activity was assessed by bio-film disruption and serum sterilization assays.Results The tail spike protein of phage vB_Kpn_HF1013 exhibited depolymerase activity and recombinant protein Dep44 specifically degraded K64 CRKP capsule.Biofilm eradication assays demonstrated that recombinant Dep44 at both 2 μg/mL and 10 μg/mL significantly disrupted bacterial bio-films relative to the control.Serum bactericidal assays showed that Dep44 exhibited synergistic activity with serum,dependent on the complement system,as Dep44 alone lacked bactericidal properties.Conclusion Dep44 effec-tively targets and degrades K64 CRKP capsule,disrupts biofilms,and enhances serum bactericidal activity,high-lighting its potential for managing K64 CRKP infections and clearing biofilms from medical devices.
