牛源肺炎克雷伯菌前噬菌体内溶素的表达与抗菌活性研究  

Expression and Antibacterial Activity Verification of Endolysin in Prophages of Klebsiella pneumoniae from Cattle

作　　者：刘初阳 胡瑞瑞 李福林 崔超文 胡圣伟[1] LIU Chuyang;HU Ruirui;LI Fulin;CUI Chaowen;HU Shengwei(College of Life Sciences,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学生命科学学院,石河子832000

出　　处：《中国畜牧兽医》2024年第9期4132-4142,共11页China Animal Husbandry & Veterinary Medicine

基　　金：新疆维吾尔自治区天山英才项目。

摘　　要：【目的】预测并分析牛源肺炎克雷伯菌携带的前噬菌体以及其携带内溶素基因的情况,探究肺炎克雷伯菌前噬菌体内溶素的体外抗菌活性,为肺炎克雷伯菌感染提供新的治疗手段。【方法】以134株牛源肺炎克雷伯菌为研究对象,通过PhiSpy软件预测牛源肺炎克雷伯菌前噬菌体,并对前噬菌体进行功能注释鉴定得到内溶素。通过构建内溶素原核表达载体,对重组蛋白进行诱导表达与纯化,并验证其在体外对肺炎克雷伯菌NMG10、GS4和ATCC 13883的裂解活性。【结果】134株牛源肺炎克雷伯菌基因组的前噬菌体序列携带率为98.5%,共预测到414个前噬菌体序列,其中肺炎克雷伯菌噬菌体共112个,其余均为其他细菌的噬菌体或者噬菌体元件。每株牛源肺炎克雷伯菌所携带的全部前噬菌体占其基因组比例约为1.4%。112个肺炎克雷伯菌噬菌体的前噬菌中共有44个前噬菌体基因组中存在编码内溶素的基因,将其命名为LyKb。内溶素LyKb对肺炎克雷伯菌NMG10、GS4和ATCC 13883均具有裂解和抑菌活性,其中对肺炎克雷伯菌NMG10的裂解及抑菌活性最强,其次是肺炎克雷伯菌GS4、ATCC 13883。【结论】134株牛源肺炎克雷伯菌普遍携带前噬菌体,且部分前噬菌体基因组中含有编码内溶素的基因。肺炎克雷伯菌前噬菌体内溶素对多种肺炎克雷伯菌具有抗菌活性。【Objective】The purpose of this experiment was to predict and analyze the prophage carried by Klebsiella pneumoniae and its carrying endolysin gene,explore the antibacterial activity of endolysin in vitro,and provide a new treatment for Klebsiae pneumoniae infection.【Method】Using 134 strains of bovine-origin Klebsiella pneumoniae as the research subjects,the prophages in bovine-origin Klebsiella pneumoniae were predicted through the PhiSpy software.Functional annotation and identification of endolysin were performed on the predicted prophages.The recombinant protein was induced and purified by constructing endolysin prokaryotic expression vector,and its lytic activity against Klebsiella pneumoniae NMG10,GS4 and ATCC 13883 was verified in vitro.【Result】The prophage sequence carriage rate in the genomes of 134 strains of bovine-origin Klebsiella pneumoniae was 98.5%,with a total of 414 predicted prophage sequences.Among them,112 belonged to Klebsiella pneumoniae phages,while the other were either phages or phage components of other bacteria.The proportion of all prophages carried by each bovine-origin Klebsiella pneumoniae genome was approximately 1.4%.A total of 44 prophage genes encoding endolysin were found in the genome of 112 prophages of Klebsiella pneumoniae,which was named LyKb.Endolysins LyKb exhibited both lytic and antibacterial activities against Klebsiella pneumoniae strains NMG10,GS4 and ATCC 13883.Among these,LyKb showed the strongest lytic and antibacterial activities against Klebsiella pneumoniae NMG10,followed by Klebsiella pneumoniae GS4 and ATCC 13883.【Conclusion】134 strains of bovine-origin Klebsiella pneumoniae commonly carried prophages,and some of the prophage genomes contained genes encoding endolysins.The endolysins of Klebsiella pneumoniae prophages had antibacterial activity against various strains of Klebsiella pneumoniae.

关 键 词：肺炎克雷伯菌 前噬菌体 内溶素 裂解 

分 类 号：S852.61[农业科学—基础兽医学]