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Biological characteristics and genome analysis of a novel phage vB_KpnP_IME279 infecting Klebsiella pneumoniae

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Abstract

Klebsiella pneumoniae (family Enterobacteriaceae) is a gram-negative bacterium that has strong pathogenicity to humans and can cause sepsis, pneumonia, and urinary tract infection. In recent years, the unreasonable use of antibacterial drugs has led to an increase in drug-resistant strains of K. pneumoniae, a serious threat to public health. Bacteriophages, viruses that infect bacteria, are ubiquitous in the natural environment. They are considered to be the most promising substitute for antibiotics because of their high specificity, high efficiency, high safety, low cost, and short development cycle. In this study, a novel phage designated vB_KpnP_IME279 was successfully isolated from hospital sewage using a multidrug-resistant strain of K. pneumoniae as an indicator. A one-step growth curve showed that vB_KpnP_IME279 has a burst size of 140 plaque-forming units/cell and a latent period of 20 min at its optimal multiplicity of infection (MOI = 0.1). Phage vB_KpnP_IME279 survives in a wide pH range between 3 and 11 and is stable at temperatures ranging from 40 to 60 °C. Ten of the 20 strains of K. pneumoniae including the host bacteria were lysed by the phage vB_KpnP_IME279, and the multilocus sequence typing and wzi typing of the 10 strains were ST11, ST37, ST375, wzi209, wzi52, and wzi72, respectively. The genome of vB_KpnP_IME279 is 42,518 bp long with a G + C content of 59.3%. Electron microscopic observation showed that the phage belongs to the family Podoviridae. BLASTN alignment showed that the genome of the phage has low similarity with currently known phages. The evolutionary relationship between phage vB_KpnP_IME279 and other Podoviridae was analyzed using a phylogenetic tree based on sequences of phage major capsid protein and indicates that the phage vB_KpnP_IME279 belongs to the Podoviridae subfamily. These data enhance understanding of K. pneumoniae phages and will help in development of treatments for multidrug-resistant bacteria using phages.

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Funding

This research was supported by a grant from the National Key Research and Development Program of China (2016YFC1202705, AWS16J020, and AWS15J006), the National Natural Science Foundation of China (81572045, 81672001, and 81621005), the Fundamental Research Funds for the Central Universities and Research Projects on Biomedical Transformation of China-Japan Friendship Hospital (No. PYBZ1820), the National Science and Technology Major Project (Grant No. 2018ZX10201001), and Project 31900489 supported by National Natural Science Foundation of China.

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Authors and Affiliations

  1. School of Public Health, Lanzhou University, Lanzhou, 730000, China

    Rongrong Zhang & Hui Liu

  2. State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 100071, China

    Feiyang Zhao, Guangqian Pei, Hang Fan, Lilan Zhangxiang, Zhiqiang Mi & Yigang Tong

  3. Beijing Advanced Innovation Center for Soft Matter Science and Engineering (BAIC-SM), College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China

    Jiuru Wang & Yigang Tong

  4. Academy of Military Medical Sciences, Beijing, 100085, China

    Taoxing Shi

Authors
  1. Rongrong Zhang
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  2. Feiyang Zhao
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  3. Jiuru Wang
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  4. Guangqian Pei
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  5. Hang Fan
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  6. Lilan Zhangxiang
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  7. Zhiqiang Mi
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  8. Taoxing Shi
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  9. Hui Liu
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  10. Yigang Tong
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Contributions

Yigang Tong, Hui Liu, Taoxing Shi, and Zhiqiang Mi conceived and designed experiments and critically evaluated the manuscript. Rongrong Zhang, Feiyang Zhao, and Jiuru Wang contributed equally to the writing of this article. Rongrong Zhang was responsible for data and sequence analyses and wrote the manuscript. Feiyang Zhao extracted the phage nucleotide and conducted the sequencing experiments. Jiuru Wang isolated and identified the phage and conducted the biological characterization experiments. Guangqian Pei helps measuring phage DNA sequences. Hang Fan and Lilan Zhangxiang help analysis the bioinformation of the phage. All authors read and approved the final manuscript

Corresponding author

Correspondence to Hui Liu.

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Zhang, R., Zhao, F., Wang, J. et al. Biological characteristics and genome analysis of a novel phage vB_KpnP_IME279 infecting Klebsiella pneumoniae. Folia Microbiol 65, 925–936 (2020). https://doi.org/10.1007/s12223-020-00775-8

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  • DOI: https://doi.org/10.1007/s12223-020-00775-8

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