牛粪中链霉菌的分离鉴定及其抗菌活性  

Isolation and Identification of Streptomyce from Cow Dung and Antimicrobial Activity of Its Substance

作　　者：尹逊哲 郭焱[2] 李文杰[1] 姜爽[2] 于秀明[3] 金宁一[4] YIN Xunzhe;GUO Yan;LI Wenjie;JIANG Shuang;YU Xiuming;JIN Ningyi(College of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China;College of Basic Medical,Changchun University of Chinese Medicine,Changchun 130117,China;Affiliated Hospital,Changchun University of Chinese Medicine,Changchun 130021,China;Institute of Military Veterinary,Academy of Military Medical Sciences,Changchun 130122,China)

机构地区：[1]长春中医药大学药学院,长春130117 [2]长春中医药大学基础医学院,长春130117 [3]长春中医药大学附属医院,长春130021 [4]军事医学科学院军事兽医研究所,长春130122

出　　处：《医药导报》2018年第1期40-43,共4页Herald of Medicine

基　　金：吉林省卫生计生科研计划(2014Z099);吉林省教育厅"十二五"科学技术研究规划项目(吉教科合字[2015]第333号);吉林省教育厅"十三五"产业化项目(JJKH20170739KJ);吉林省科技厅自然科学基金资助项目(20160101224JC)

摘　　要：目的分离鉴定牛粪中的链霉菌,观察其抗菌活性。方法采用稀释涂布法从牛粪中分离菌株,通过固定琼脂块法以金黄色葡萄球菌、大肠埃希菌为指示菌筛选抗菌活性较强的菌株,并对菌种进行生理生化鉴定及16S r DNA序列测定;采用纸片扩散法测定活性菌株发酵液抗菌活性,菌株发酵液抗菌活性物质进行水饱和正丁醇萃取,并通过Molish反应、双缩脲反应及茚三酮反应进行抗菌活性物质定性鉴别。结果从牛粪中分离出8株菌株,并筛选出1株抗菌活性较强菌株B5-2,鉴定为链霉菌。活性菌株发酵液对金黄色葡萄球菌、大肠埃希菌、费氏柠檬酸杆菌、阴沟肠杆菌、肺炎克雷伯菌、产气肠杆菌均具有较强抑菌作用。抑菌结果显示,菌株B5-2抑菌活性物质均为水溶性物质。Molish反应、双缩脲反应及茚三酮反应结果显示,菌株B5-2的抑菌活性物质均初步鉴定为糖类和蛋白质类。结论筛选分离的链霉菌对临床致病菌有较强的抑制作用。ObjectiveTo isolate and identificate Streptomyce strain from cow dung and observe its antimicrobial activity.MethodsStrains were isolated from cow dung by dilution coating method.Strong antibacterial strains were screened out by agar block method with fixed Staphylococcus aureus and Escherichia coli as indicative bacteria.The strains were identified based on physiological and biochemical characteristics as well as16S rDNA gene sequence analysis.Active antibacterial fermentation broth substance was determined by disk diffusion method,and antibacterial active substance of strains fermentation broth was extracted by water saturated n butanol.Antibacterial substance of strains was identified by Molish reaction,biuret test and ninhydrin reaction.ResultsEight strains were isolated from cow dung and one strong antibacterial strain was screened out and named B5-2,identified as Streptomyces.The results showed that the strain had the highly antibacterial effect on Staphylococcus aureus,Escherichia coli,Citrobacter freundii,Enterobacter cloacae,Klebsiella pneumoniae,Enterobacter aerogenes.The strain antibacterial active substance of fermentation broth preliminary analysis showed that strong antibacterial active substance of B5-2was the water soluble substance.Antibacterial substance of B5-2was preliminarily identified as glycoside and protein by Molish reaction,biuret test and ninhydrin reaction.ConclusionThe strain isolated have a strong inhibition effect on clinical pathogenic bacteria in clinical practice.

关 键 词：链霉菌 牛粪 鉴定 抗菌活性 

分 类 号：R978.12[医药卫生—药品] R965[医药卫生—药学]