Characterization of bacteriophage BUCT631 lytic for K1 <i>Klebsiella pneumoniae</i> and its therapeutic efficacy in <i>Galleria mellonella</i> larvae

Pengjun Han; Mingfang Pu; Yahao Li; Huahao Fan; Yigang Tong

doi:10.1016/j.virs.2023.07.002

October 2023

Pengjun Han, Mingfang Pu, Yahao Li, Huahao Fan and Yigang Tong. Characterization of bacteriophage BUCT631 lytic for K1 Klebsiella pneumoniae and its therapeutic efficacy in Galleria mellonella larvae[J]. Virologica Sinica, 2023, 38(5): 801-812. doi: 10.1016/j.virs.2023.07.002

Citation: Pengjun Han, Mingfang Pu, Yahao Li, Huahao Fan, Yigang Tong. Characterization of bacteriophage BUCT631 lytic for K1 Klebsiella pneumoniae and its therapeutic efficacy in Galleria mellonella larvae .VIROLOGICA SINICA, 2023, 38(5) : 801-812. http://dx.doi.org/10.1016/j.virs.2023.07.002

K1型肺炎克雷伯菌噬菌体BUCT631的鉴定及其对大蜡螟幼虫的治疗效果评估

韩鹏军 ^a ,
蒲明芳 ^a ,
李亚昊 ^b ,
范华昊 ^a,, ,
童贻刚 ^a,b,,

北京化工大学, 生命科学与技术学院

通讯作者： 范华昊, fanhuahao@mail.buct.edu.cn; 童贻刚, tongyigang@mail.buct.edu.cn
收稿日期： 2023-04-09
录用日期： 2023-07-03

摘要

由多重耐药性肺炎克雷伯菌（K. pneumoniae）引起的严重感染凸显了对这种病原体具有活性的新疗法的需求。噬菌体疗法是治疗多重耐药性肺炎克雷伯菌感染的一种替代治疗方法。在此，我们报告了一株新的噬菌体BUCT631，它能特异性地裂解K1荚膜类型的肺炎克雷伯菌。生理特征分析表明，噬菌体BUCT631能迅速吸附在肺炎克雷伯菌的表面并形成明显的晕环，它具有相对有利的热稳定性（4-50℃）和pH耐受性（pH=4-12）。此外，噬菌体BUCT631的最佳感染复数（MOI）为0.01，爆发量约为303 PFU/cell。基因组分析表明，噬菌体BUCT631具有双链DNA（总长度为44,812 bp），G+C含量为54.1%，基因组包含57个开放阅读框（ORFs），没有毒力或抗生素抗性相关基因。根据系统发育分析，噬菌体BUCT631可以被归入Slopekvirinae亚科Drulisvirus属中的一个新种。最重要的是，噬菌体BUCT631可以在体外2小时内迅速抑制肺炎克雷伯菌的生长，并在体内将感染肺炎克雷伯菌的大蜡螟幼虫的存活率从10%显著提高到90%。这些研究表明，噬菌体BUCT631具有很好的开发潜力，是控制和治疗多重耐药性K1肺炎克雷伯菌感染的安全替代品。
- 噬菌体BUCT631
- , 肺炎克雷伯菌
- , 基因组分析
- , 噬菌体治疗
- , 大蜡螟幼虫

Characterization of bacteriophage BUCT631 lytic for K1 Klebsiella pneumoniae and its therapeutic efficacy in Galleria mellonella larvae

Pengjun Han ^a ,
Mingfang Pu ^a ,
Yahao Li ^b ,
Huahao Fan ^a,, ,
Yigang Tong ^a,b,,

a. College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, China;
b. Beijing Advanced Innovation Center for Soft Matter Science and Engineering, Beijing University of Chemical Technology, Beijing, 100029, China

Corresponding author: Huahao Fan, fanhuahao@mail.buct.edu.cn Yigang Tong, tongyigang@mail.buct.edu.cn
Received Date: 09 April 2023
Accepted Date: 03 July 2023

Abstract

Severe infections caused by multidrug-resistant Klebsiella pneumoniae (K. pneumoniae) highlight the need for new therapeutics with activity against this pathogen. Phage therapy is an alternative treatment approach for multidrug-resistant K. pneumoniae infections. Here, we report a novel bacteriophage (phage) BUCT631 that can specifically lyse capsule-type K1 K. pneumoniae. Physiological characterization revealed that phage BUCT631 could rapidly adsorb to the surface of K. pneumoniae and form an obvious halo ring, and it had relatively favorable thermal stability (4-50 °C) and pH tolerance (pH=4-12). In addition, the optimal multiplicity of infection (MOI) of phage BUCT631 was 0.01, and the burst size was approximately 303 PFU/cell. Genomic analysis showed that phage BUCT631 has double-stranded DNA (total length of 44,812 bp) with a G + C content of 54.1%, and the genome contains 57 open reading frames (ORFs) and no virulence or antibiotic resistance related genes. Based on phylogenetic analysis, phage BUCT631 could be assigned to a new species in the genus Drulisvirus of the subfamily Slopekvirinae. In addition, phage BUCT631 could quickly inhibit the growth of K. pneumoniae within 2 h in vitro and significantly elevated the survival rate of K. pneumoniae infected Galleria mellonella larvae from 10% to 90% in vivo. These studies suggest that phage BUCT631 has promising potential for development as a safe alternative for control and treatment of multidrug-resistant K. pneumoniae infection.
- Bacteriophage BUCT631
- , Klebsiella pneumoniae
- , Genomic analysis
- , Phage therapy
- , Galleria mellonella

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