抗分枝杆菌海洋微生物的分离与鉴定  

Isolation and identification of marine bacteria with bacteriostasis to Mycobacterium smegmatis MC;155

作　　者：关艳[1] 蒙建州[1] 刘忆霜[1] 肖春玲[1] 杨延辉 GUAN Yan;MENG Jian-zhou;LIU Yi-shuang;XIAO Chun-ling;YANG Yan-hui(NHC Key Laboratory of Biotechnology of Antibiotics,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China;Ningxia Key Laboratory of Prevention and Control of Common Infectious Diseases,Department of Pathogenic Biology and Medical Immunology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所国家新药(微生物)筛选室,北京100050 [2]宁夏医科大学基础医学院病原生物学与医学免疫学系宁夏回族自治区常见传染病防治重点实验室,银川750004

出　　处：《中国医药生物技术》2022年第1期4-11,共8页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(81560604);中国科学院“西部之光”人才培养计划(XAB2019AW11);中国医学科学院医学与健康科技创新工程(2021-12M-1-028);宁夏回族自治区重点研发计划(2021BEG 03072)。

摘　　要：目的从海洋微生物中发现具有抗分枝杆菌活性的细菌,并对其进行鉴定。方法从印度洋热带地区的海底沉积物中分离海洋微生物。通过纸片法活性测定,发现了一株具有抗耻垢分枝杆菌活性的细菌。光学显微下观察细菌形态和菌落。测定细菌的理化特性,提取其基因组,荧光法测定基因组的T_(m)值,扩增其16S rDNA并进行测序。将序列提交NCBI和EzTaxon网站进行搜索比对,使用MEGA 7.0软件构建系统进化树。结果分离得到的菌株IMBGY10-46的发酵产物具有抗耻垢分枝杆菌活性;其与Pseudomonas sabulinigriJ64的16S rDNA相似性最高,但菌体形态和基因组的Tm值相差较大,并且过氧化氢酶试验呈阴性。将该菌株命名为假单胞菌属新菌株。该细菌的16S rDNA序列提交GenBank,收录号为JQ362457。结论从海底沉积物中获得了具有抗耻垢分枝杆菌作用的海洋微生物新菌株。Objective We aim to identify novel anti-mycobacterial microorganisms from marine microbes.Methods Marine microorganisms were separated from the samples of seafloor sediments from the Indian Ocean tropical region. A paper disk method was used to determine the anti-mycobacteria activity, whereas light microscopy was utilized to learn the morphologies of bacteria and colony. The physicochemical properties of IMBGY10-46 were measured. The genome of IMBGY10-46 was extracted by genomic purification kit. In addition, fluorescence spectrometry was harnessed to determine the T_(m) value of bacterial genome. 16 S rDNA was amplified by PCR and its sequence was subjected to BLAST in NCBI and EzTaxon. A phylogenetic tree was built by MEGA 7.0.Results Anti-mycobacterial activity was discovered in the fermentative liquid of IMBGY10-46, whose 16 S rDNA had the highest homology with that of Pseudomonas sabulinigri J64. Nevertheless, there were diverse in morphology and in Tm value of genome.The strain IMBGY10-46 was found to be catalase-negative. To this end, the 16 S rDNA sequence of IMBGY10-46 was deposited in GenBank(Accession No. JQ362457).Conclusion A new marina bacteria isolated from the seafloor sediment can produce secondary metabolites against Mycobacterium smegmatis.

关 键 词：海洋微生物 次级代谢产物 抗菌活性 耻垢分枝杆菌 16S rDNA 

分 类 号：R915[医药卫生—微生物与生化药学]