Identification of a phage-derived depolymerase specific for KL47 capsule of <em>Klebsiella pneumoniae</em> and its therapeutic potential in mice

Min Li; Hui Wang; Long Chen; Genglin Guo; Pei Li; Jiale Ma; Rong Chen; Hong Du; Yuqing Liu; Wei Zhang

doi:10.1016/j.virs.2022.04.005

August 2022

Min Li, Hui Wang, Long Chen, Genglin Guo, Pei Li, Jiale Ma, Rong Chen, Hong Du, Yuqing Liu and Wei Zhang. Identification of a phage-derived depolymerase specific for KL47 capsule of Klebsiella pneumoniae and its therapeutic potential in mice[J]. Virologica Sinica, 2022, 37(4): 538-546. doi: 10.1016/j.virs.2022.04.005

Citation: Min Li, Hui Wang, Long Chen, Genglin Guo, Pei Li, Jiale Ma, Rong Chen, Hong Du, Yuqing Liu, Wei Zhang. Identification of a phage-derived depolymerase specific for KL47 capsule of Klebsiella pneumoniae and its therapeutic potential in mice .VIROLOGICA SINICA, 2022, 37(4) : 538-546. http://dx.doi.org/10.1016/j.virs.2022.04.005

对肺炎克雷伯菌KL47型荚膜特异的噬菌体源解聚酶的鉴定及其对小鼠的治疗潜力

李敏 ^a,b,c ,
王惠 ^a,b,c ,
陈龙 ^d ,
郭庚霖 ^a,b,c ,
李培 ^a,b,c ,
马家乐 ^a,b,c ,
陈蓉 ^e ,
杜鸿 ^f ,
刘玉庆 ^g ,
张炜 ^a,b,c,,

1 南京农业大学动物医学院, 南京, 210095, 中国;
2 动物细菌学重点实验室, 农业部, 南京 210095, 中国;
3 教育部动物卫生与食品安全国际合作联合实验室, 南京 210095, 中国;
4 苏州大学附属张家港医院, 张家港 215600, 中国;
5 江苏省农业科学院兽医研究所, 南京 210014, 中国;
6 苏州大学第二附属医院临床检验科, 苏州 215004, 中国;
7 山东省农业科学院畜牧兽医研究所, 济南 250100, 中国

通讯作者： 张炜, vszw@njau.edu.cn
收稿日期： 2021-10-14
录用日期： 2022-04-28

摘要

肺炎克雷伯菌是引起全球多重耐药感染的主要病原体之一。因此，迫切需要制定有效的防控策略。噬菌体源解聚酶常存在于尾纤维蛋白或尾刺蛋白中，据报道具有抗生物被膜活性。本研究中，从污水中分离的噬菌体P560对KL47型肺炎克雷伯菌具有特异性，其噬菌斑周围扩大的晕圈暗示P560可能编码一种解聚酶。从噬菌体P560中预测到解聚酶ORF43，命名为P560dep，我们对其进行了表达、纯化、鉴定以及评估酶活性和特异性。结果表明荚膜解聚酶P560dep可以消化KL47型肺炎克雷伯菌表面的荚膜多糖，并且P560dep的解聚谱与噬菌体P560的宿主范围相匹配，即KL47型肺炎克雷伯菌。结晶紫染色结果显示，P560dep能显著抑制生物被膜的形成。此外，腹腔注射单剂量（50µg/只）的解聚酶P560dep可保护90%-100%的小鼠免受KL47型耐碳青霉烯类肺炎克雷格菌感染前后的致命攻击。感染KL47型肺炎克雷伯菌的小鼠经P560dep治疗后肺、肝组织病理改变均有所减轻。这也证明解聚酶P560dep作为一种有吸引力的抗毒力制剂代表了一种有前景的抗菌治疗工具。
- 噬菌体源解聚酶
- , 耐碳青霉烯类肺炎克雷伯菌
- , 荚膜型
- , 生物被膜
- , 抗菌疗法

Identification of a phage-derived depolymerase specific for KL47 capsule of Klebsiella pneumoniae and its therapeutic potential in mice

Min Li ^a,b,c ,
Hui Wang ^a,b,c ,
Long Chen ^d ,
Genglin Guo ^a,b,c ,
Pei Li ^a,b,c ,
Jiale Ma ^a,b,c ,
Rong Chen ^e ,
Hong Du ^f ,
Yuqing Liu ^g ,
Wei Zhang ^a,b,c,,

a College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China;
b Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing, 210095, China;
c MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China;
d Department of Clinical Laboratory, Zhangjiagang Hospital Affiliated to Soochow University, Zhangjiagang, 215600, China;
e Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing, 210014, China;
f Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, 215004, China;
g Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China

Corresponding author: Wei Zhang, vszw@njau.edu.cn
Received Date: 14 October 2021
Accepted Date: 28 April 2022

Abstract

Klebsiella pneumoniae is one of the major pathogens causing global multidrug-resistant infections. Therefore, strategies for preventing and controlling the infections are urgently needed. Phage depolymerase, often found in the tail fiber protein or the tail spike protein, is reported to have antibiofilm activity. In this study, phage P560 isolated from sewage showed specific for capsule locus type KL47 K. pneumoniae, and the enlarged haloes around plaques indicated that P560 encoded a depolymerase. The capsule depolymerase, ORF43, named P560dep, derived from phage P560 was expressed, purified, characterized and evaluated for enzymatic activity as well as specificity. We reported that the capsule depolymerase P560dep, can digest the capsule polysaccharides on the surface of KL47 type K. pneumoniae, and the depolymerization spectrum of P560dep matched to the host range of phage P560, KL47 K. pneumoniae. Crystal violet staining assay showed that P560dep was able to significantly inhibit biofilm formation. Further, a single dose (50 μg/mouse) of depolymerase intraperitoneal injection protected 90%–100% of mice from lethal challenge before or after infection by KL47 carbapenem-resistant K. pneumoniae. And pathological changes were alleviated in lung and liver of mice infected by KL47 type K. pneumoniae. It is demonstrated that depolymerase P560dep as an attractive antivirulence agent represents a promising tool for antimicrobial therapy.
- Bacteriophage-derived depolymerase
- , Carbapenem-resistant Klebsiella pneumoniae
- , Capsular type
- , Biofilm
- , Antimicrobial therapy

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10.1016j.virs.2022.04.005-ESM.docx