新型R型噬菌体尾样细菌素对MRSA的抗菌活性研究  

Antimicrobial activity of a novel R-type phage tail-like bacteriocin against MRSA

作　　者：唐伟 刘颖 唐震海 唐颖 李昕[1] 姚杰[1] 李维祖 徐元宏[6] 周强[1] TANG Wei;LIU Ying;TANG Zhen-hai;TANG Ying;LI Xin;YAO Jie;LI Wei-zu;XU Yuan-hong;ZHOU Qiang(Dept of Clinical Laboratory,the Second Affiliated Hospital of Anhui Medical University,Hefei 230601,China;High Magnetic Field Laboratory,Hefei Institutes of Physical Science,Chinese Academy of Sciences,Hefei 230031,China;Center for Scientific Research of Anhui Medical University,Hefei 230032,China;School of Clinical Medicine,Wannan Medical College,Wuhu Anhui 241002,China;Dept of Pharmacology,School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China;Dept of Clinical Laboratory,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China)

机构地区：[1]安徽医科大学第二附属医院检验科,安徽合肥230601 [2]中国科学院合肥物质科学研究院强磁场科学中心,安徽合肥230031 [3]安徽医科大学科研实验中心,安徽合肥230032 [4]皖南医学院临床医学院,安徽芜湖241002 [5]安徽医科大学基础医学院药理学教研室,安徽合肥230032 [6]安徽医科大学第一附属医院检验科,安徽合肥230022

出　　处：《中国药理学通报》2024年第9期1760-1765,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82102460);安徽医科大学第二附属医院临床研究培育计划(No 2021LCZD13);安徽医科大学校科研基金项目(No 2022xkj197)。

摘　　要：目的探索阴沟肠杆菌SHAMU191747所分泌的新型R型噬菌体尾样细菌素(phage tail-like bacteriocin,PTLB)对耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)的抗菌活性。方法琼脂平板拮抗试验和肉汤微量发酵试验,检测阴沟肠杆菌SHAMU191747对MRSA的拮抗活性。超高速离心法和密度梯度离心法,制备阴沟肠杆菌SHAMU191747发酵上清液粗提物。使用透射电镜检测粗提物中有无新型R型PTLB。琼脂平板点种法,验证粗提物对MRSA的抗菌活性。采用SDS-PAGE电泳检测新型R型PTLB的分子量。结果阴沟肠杆菌SHAMU191747分泌新型R型PTLB,对MRSA具有强拮抗效应。新型R型PTLB的分子量约35 ku,能够高效杀伤MRSA;尾鞘未收缩的功能性分子的物理尺寸为(142.7±4.3)×(13.8±0.6)nm,尾鞘收缩的非功能性分子的物理尺寸为(57.7±1.2)×(20.8±1.5)nm。结论阴沟肠杆菌SHAMU191747产生的新型R型PTLB能够高效杀伤MRSA,具有开发成新型抗菌药物的潜力,临床应用前景很大。Aim To explore the antimicrobial activity of a novel R-type phage tail-like bacteriocin(PTLB)secreted by Enterobacter cloacae SHAMU191747 against methicillin-resistant Staphylococcus aureus(MRSA).Methods Antagonistic activity of E.cloacae SHAMU191747 against MRSA was detected by LB agar plate antagonistic test and LB broth micro-fermentation test.The crude extract of fermentation supernatant of E.cloacae SHAMU191747 was prepared by ultra-high-speed centrifugation and density gradient centrifugation.The novel R-type PTLB in the crude extract was detected by transmission electron microscopy.The antimicrobial activity of the crude extract against MRSA was verified by LB agar plate spot-seeding method.The molecular weight of the novel R-type PTLB was detected by SDS-PAGE electrophoresis.Results E.cloacae SHAMU191747 secreted a novel R-type PTLB,and had a strong antagonistic effect on MRSA.The novel R-type PTLB had a molecular weight of approximately 35 ku and could efficiently kill MRSA.The physical dimensions of its tail-sheath-uncontracted functional molecules were(142.7±4.3)×(13.8±0.6)nm,and those of the tail-sheath-contracted nonfunctional molecules were(57.7±1.2)×(20.8±1.5)nm.Conclusions The novel R-type PTLB produced by E.cloacae SHAMU191747 can efficiently kill MRSA,and has the potential to be developed into a novel antimicrobial drug with great prospects for clinical application.

关 键 词：阴沟肠杆菌 细菌素 噬菌体尾样细菌素 金黄色葡萄球菌 耐甲氧西林金黄色葡萄球菌 抗菌活性 

分 类 号：R378.11[医药卫生—病原生物学] R378.2[医药卫生—基础医学] R978.1