摘要
为了解抗菌肽RL-8的抑菌活性及抗大肠杆菌的作用机制,应用生物信息学工具分析了抗菌肽RL-8的理化特征和二级结构,通过微量稀释法测定抗菌肽RL-8对部分细菌和真菌的抑菌活性,采用菌落计数法测定抗菌肽RL-8对大肠杆菌CVCC1568的杀菌动力学,利用分光光度法分析抗菌肽RL-8对大肠杆菌CVCC1568细胞壁和细胞膜的破坏作用。结果表明:抗菌肽RL-8为阳离子型α螺旋形两亲性多肽,对革兰阴性菌(大肠杆菌、沙门菌、猪胸膜肺炎放线杆菌、绿脓杆菌)的最小抑菌浓度(MIC)为6.25~200.00 mg/L,对革兰阳性菌(金黄色葡萄球菌、枯草芽孢杆菌、屎肠球菌、短小芽孢杆菌)的MIC为3.12~400.00 mg/L,对白色念珠菌的MIC为200.00 mg/L。抗菌肽RL-8能在60 min内杀灭99.99%的大肠杆菌CVCC1568,可破坏大肠杆菌的细胞壁和细胞膜,造成胞内紫外吸收物质泄露。文章研究结果为抗菌肽RL-8的进一步研究和临床应用提供参考。
To understand antimicrobial activity and action mechanism against Escherichia coli(E. coli) of antibacterial peptide RL-8, bioinformatics tools were used to analyze the physical and chemical characteristics and secondary structure of antimicrobial peptide RL-8, the microdilution method was used to determine bacteriostatic activity of antibacterial peptide RL-8 against several strains of bacteria and fungi, the colony counting method was used to determine the bactericidal dynamics of antibacterial peptide RL-8 against E. coli CVCC1568, and spectrophotometric method was used to analyze the damage influence of antibacterial peptide RL-8 on cell wall and cell membrane of E. coli CVCC1568. The results showed that antibacterial peptide RL-8 was a kind of cationic α helix amphiphilic peptides, the minimum inhibitory concentration(MIC) of antibacterial peptide RL-8 against several gram-negative bacteria(E.coli, Salmonella, Actinobacillus pleuropneumonia, Pseudomonas aeruginosa) was 6.25~200.00 mg/L, the MIC of antibacterial peptide RL-8 against several Gram-positive bacteria(Staphylococcus aureus, Bacillus subtilis, Enterococcus faecium, Bacillus pumilus) was 3.12~400.00 mg/L, the MIC of antibacterial peptide RL-8 against Candida albicans was 200.00 mg/L. The antimicrobial peptide RL-8 could kill 99.99% of the E. coli CVCC1568 within 60 min, and could damage the cell wall and cell membrane of E. coli CVCC1568 which causes the leakage of intracellular ultraviolet absorbent materials. These findings provided reference for further study and clinical application of the antimicrobial peptide RL-8.
作者
李杰
陈文杰
杭柏林
刘保国
宁春妹
秦爱建
钱琨
Li Jie;Chen Wenjie;Hang Bolin;Liu Baoguo;Ning Chunmei;Qin Aijian;Qian Kun(Ministry of Education Key Lab for Avian Preventive Medicine,Key Laboratory of Jiangsu Preventive Veterinary Medicine,Colelge of Veterinary Medicine,Yangzhou University,Jiangsu Yangzhou 225009;Xnixiang Center of Monitor and Inspection for Quality of Livestokc Production,Henan Xinxiang 453003;College of Animal Science and Veterinary Medicine,Henan Institute of Science and Technology,Henan Xinxiang 453003)
出处
《现代畜牧兽医》
2020年第9期5-10,共6页
Modern Journal of Animal Husbandry and Veterinary Medicine
基金
教育部禽类预防医学重点实验室/江苏省动物预防医学重点实验室开放课题(YF201802)
河南省自然科学基金项目(182300410031)
河南科技学院高层次人才启动项目(2014020)
河南科技学院大学生“百农英才”创新项目(BNYC2018-2-05)河南省大学生创新创业训练计划项目(S201910467012)。
关键词
抗菌肽
抑菌活性
大肠杆菌
胞内物质泄露
Antimicrobial peptides
Bacteriostatic activity
Escherichia coli
Leakage of intracellular substances
