CAJ | 학술논문

本研究旨在通过分离粪肠球菌裂解性噬菌体并分析其生物学特性，为噬菌体疗法应用于肠球菌感染提供依据。以粪肠球菌为宿主菌，用双层琼脂法从环境污水中分离出1株裂解性噬菌体（命名为φEn‐ZZ8），对其生物学特性进行初步分析。透射电镜观察发现，φEn‐ZZ8头部呈二十面体立体对称，直径约40 nm ；尾部呈丝状，长约200 nm。吸附实验表明，在10 min内超过90％的噬菌体可吸附至宿主菌细胞。一步生长曲线显示，φEn‐ZZ8的潜伏期为15 min ，爆发期为40 min ，爆发量为150 pfu／cell。φEn‐ZZ8基因组对内切酶Hind Ⅲ、EcoR Ⅴ、NdeⅠ、PstⅠ、X baⅠ和 SacⅡ敏感；基因组大小约为42 kb。十二烷基硫酸钠‐聚丙烯酰胺凝胶电泳至少可见14条蛋白带，相对分子质量为10000～130000。φEn‐ZZ8对26株粪肠球菌的裂解率达30．8％，其裂解活性在pH 5～8和50～70℃条件下仍稳定。结果提示，φEn‐ZZ8是一种裂解性强、裂解谱较宽、对温度具有耐受性的裂解性噬菌体，有望成为抗耐药性肠球菌感染的噬菌体生物制剂。
본연구지재통과분리분장구균렬해성서균체병분석기생물학특성，위서균체요법응용우장구균감염제공의거。이분장구균위숙주균，용쌍층경지법종배경오수중분리출1주렬해성서균체（명명위φEn‐ZZ8），대기생물학특성진행초보분석。투사전경관찰발현，φEn‐ZZ8두부정이십면체입체대칭，직경약40 nm ；미부정사상，장약200 nm。흡부실험표명，재10 min내초과90％적서균체가흡부지숙주균세포。일보생장곡선현시，φEn‐ZZ8적잠복기위15 min ，폭발기위40 min ，폭발량위150 pfu／cell。φEn‐ZZ8기인조대내절매Hind Ⅲ、EcoR Ⅴ、NdeⅠ、PstⅠ、X baⅠ화 SacⅡ민감；기인조대소약위42 kb。십이완기류산납‐취병희선알응효전영지소가견14조단백대，상대분자질량위10000～130000。φEn‐ZZ8대26주분장구균적렬해솔체30．8％，기렬해활성재pH 5～8화50～70℃조건하잉은정。결과제시，φEn‐ZZ8시일충렬해성강、렬해보교관、대온도구유내수성적렬해성서균체，유망성위항내약성장구균감염적서균체생물제제。
This study aims to isolate lytic bacteriophages from Enterococcus faecalis (E . faecalis) ,analyze its biological characteristics and to establish the research basis for a phage‐based therapy against Enterococcus infections .A lytic bacteriophage (named φEn‐ZZ8) was isolated from the sewage using double‐agar method and its basic biological characteristics were analyzed .The results showed that φEn‐ZZ8 had an icosahedral head of 40 nm in diameter and a filamentous tail of 200 nm in length .The adsorption rate of φEn‐ZZ8 reached 90% in 10 min .One‐step growth showed that φEn‐ZZ8 had a short latency period of 15 min ,a rise period of 40 min ,and a burst size of 150 pfu/cell .The genome of φEn‐ZZ8 was sensitive to HindIII , EcoRV ,NdeI ,PstI ,Xba I and Sac II ,and the size of the genome was estimated to be 42 kb .Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE ) revealed that at least 14 proteins were observed on the gel , ranging from 11 000 to 130 000 . The bacteriophage specifically infected and lysed 30 .8% E . faecalis isolates .TheφEn‐ZZ8 also exhibited stability to the wide range of pH and temperature . The data suggest that phage φEn‐ZZ8 may be a candidate therapeutic agent to control Enterococcus infections .