塔克拉玛干沙生植物来源放线菌抗铜绿假单胞菌活性的筛选及菌株38-7L-1活性产物的研究  

Endophytic actinomycetes from psammophytes of Taklamakan Desert against Pseudomonas aeruginosa and study on bioactive product of strain 38-7L-1

作　　者：陈川[1] 刘佳萌[1] 蒋忠科[1] 李小俊[1,2] 刘少伟[1] 庹利 戴素娟[1] 杨桂柳 王飞飞[1] 黄大林[4] 姜明国[3] 孙承航[1] 

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,北京100050 [2]河北北方学院医学检验学院,张家口075000 [3]广西民族大学海洋与生物技术学院广西高校微生物与植物资源利用重点实验室,南宁530008 [4]桂林医学院,桂林541004

出　　处：《中国抗生素杂志》2015年第2期81-87,115,共8页Chinese Journal of Antibiotics

基　　金：国家自然科学基金(No.81172963;No.81373308和No.31260004);北京市自然科学基金(No.7133249);广西高校科研项目(No.201203YB117);广西自然基金(No.2012GXNSFAA053038);高等学校博士学科点专项科研基金博导类项目(No.20111106110032);高等学校博士学科点专项科研基金新教师类项目(No.20131106120027)

摘　　要：目的筛选来自塔克拉玛干沙漠沙生植物对铜绿假单胞菌具有抑制活性的内生放线菌菌株,对拟诺卡菌属菌株38-7L-1发酵液中活性化合物进行分析预测、分离纯化和结构验证。方法采用琼脂平板法进行抗菌活性筛选;基于16S r RNA基因序列对菌株38-7L-1进行初步分类鉴定;基于菌株38-7L-1 PKS I基因的KS域序列分析结果 ,结合活性峰的液-质联用数据,比对微生物天然产物数据库,预测活性化合物201的结构;通过色谱方法纯化化合物201;结合光谱数据分析及文献比对,确定和验证化合物201的结构。结果从81株沙生植物内生放线菌中获得7株活性菌株,包括链霉菌属菌株4株、考克菌属菌株1株、多形孢菌属菌株1株和拟诺卡菌属菌株1株。其中菌株38-7L-1活性最强并具有I型聚酮合酶基因,16S r RNA基因序列分析表明该菌株与Nocardiopsis alba DSM 43377T(X97883)的相似率为100%。菌株3 8-7L-1抗铜绿假单胞菌活性次级代谢产物,化合物201为十六元大环内酯类化合物,蔷薇霉素。结论 首次从拟诺卡菌中分离到常见于小单孢菌的蔷薇霉素,多策略组合对次级代谢产物结构预测具有指导意义。Objective To screen endophytic actinomycetes isolated from psammophytes collected in Taklamakan Desert with inhibitory activity against Pseudomonas aeruginosa, and predict, isolate, purify and identificate the structure of bioactive component produced by Nocardiopsis strain 38-7L-1. Methods The fermentation broths of 81 strains were screened by agar disc diffusion, strain 38-7L-1 was identified on the basis of sequence analysis of 16S rRNA gene, a bioactive secondary metabolite named as compound 201 produced by the strain 38-7L-1 was predicted by comprehensive analysis of type I PKS gene, UV and MS data provided by HPLC- DAD-MS and microbial product database; compound 201 was purified by different chromatography and its chemical structure was determined by spectral data analysis and comparison with that of published data. Results 7 strains including 4 Streptomyces strains and 3 rare actinomycetes strains were obtained as positive strains, among them, Nocardiopsis sp. 38-7L-1 showed the strongest inhibitory activity against Pseudomonas aeruginosa; predicted bioactive component produced by the strain was purified and proved to be rosamicin, a known 16-membered macrolide. Conclusion Combination of different strategy involved in discovery of antibiotic has great significance for dereplication of known antibiotic and finding new antibiotics. Rosamicin, normally produced by the genus Micromonospra, was found for the first time in the fermentation broth ofNocardiopsis sp.

关 键 词：塔克拉玛干沙漠 植物内生放线菌 铜绿假单胞菌 拟诺卡菌属 蔷薇霉素 

分 类 号：R378.991[医药卫生—病原生物学]