Protective and therapeutic application of the depolymerase derived from a novel KN1 genotype of Klebsiella pneumoniae bacteriophage in mice

Can Wang; Puyuan Li; Wenkai Niu; Xin Yuan; Huiying Liu; Yong Huang; Xiaoping An; Hang Fan; Lilan Zhangxiang; Liyuan Mi; Jing Zheng; Yannan Liu; Yigang Tong; Zhiqiang Mi; Changqing Bai

doi:10.1016/j.resmic.2019.01.003

Protective and therapeutic application of the depolymerase derived from a novel KN1 genotype of Klebsiella pneumoniae bacteriophage in mice

Res Microbiol. 2019 Apr-May;170(3):156-164. doi: 10.1016/j.resmic.2019.01.003. Epub 2019 Feb 2.

Authors

Can Wang¹, Puyuan Li², Wenkai Niu³, Xin Yuan⁴, Huiying Liu⁵, Yong Huang⁶, Xiaoping An⁷, Hang Fan⁸, Lilan Zhangxiang⁹, Liyuan Mi¹⁰, Jing Zheng¹¹, Yannan Liu¹², Yigang Tong¹³, Zhiqiang Mi¹⁴, Changqing Bai¹⁵

Affiliations

¹ Department of Respiratory and Critical Care Medicine, 307 Hospital of PLA, 307 Clinical College, Anhui Medical University, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
² Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
³ Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
⁴ Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
⁵ Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
⁶ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
⁷ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
⁸ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
⁹ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
¹⁰ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
¹¹ Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
¹² Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
¹³ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
¹⁴ State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].
¹⁵ Department of Respiratory and Critical Care Diseases, 307th Hospital of PLA, No. 8 Dongda Street, Fengtai District, Beijing, 100071, China. Electronic address: [email protected].

PMID: 30716390
DOI: 10.1016/j.resmic.2019.01.003

Abstract

Klebsiella pneumoniae is one of the major Gram-negative bacterial pathogens causing hospital-acquired multidrug-resistant infections, and the antimicrobial treatment options are scarce. The lack of available antimicrobials has prompted the development of alternative strategies for the treatment of these infections. In this study, a K. pneumoniae bacteriophage (vB_KpnP_IME321) targeting a KN1 capsular type strain, Kp409, was isolated, characterized and sequenced. This bacteriophage has a latent period of 20 min and a burst size of approximately 410 pfu/cell. It contained 49 predicted open reading frames, of which ORF42 was identified as encoding the putative capsule depolymerase. The enzyme expressed and purified in the Escherichia coli BL21 system, namely Dp42, could depolymerize the capsular polysaccharide of Kp409 and form translucent halos on the plates. The phage-encoded depolymerase could increase the inhibitory effect of serum on the growth of bacteria in vitro. Pre-treated with Dp42 rescued 100% of mice following lethal Kp409 challenge, and administration of this enzyme after infection significantly increased survival rates of infected mice in the animal experiment. In conclusion, the phage-encoded depolymerase Dp42 represents a potential alternative strategy for controlling infections mediated by K. pneumoniae expressing the KN1 capsular polysaccharide.

Keywords: Bacteriophage; Depolymerase; KN1 capsule type; Klebsiella pneumoniae; Therapeutic effects.

MeSH terms

Animals
Anti-Bacterial Agents / isolation & purification*
Anti-Bacterial Agents / therapeutic use*
Bacterial Capsules / metabolism
Bacteriophages / enzymology*
Bacteriophages / genetics
Bacteriophages / growth & development
Disease Models, Animal
Genome, Viral
Glycoside Hydrolases / genetics
Glycoside Hydrolases / isolation & purification*
Glycoside Hydrolases / therapeutic use*
Klebsiella Infections / prevention & control*
Klebsiella pneumoniae / drug effects*
Mice
Open Reading Frames
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Recombinant Proteins / therapeutic use
Sequence Analysis, DNA
Survival Analysis

Substances

Anti-Bacterial Agents
Recombinant Proteins
Glycoside Hydrolases
capsular-polysaccharide galactohydrolase