摘要
目的建立氨基戊二烯环脂肽类化合物(APD-CLDs)基因簇携带菌株的筛选策略,从实验室建立的放线菌菌种库中筛选并分析APD-CLDs基因簇携带菌株。方法采用PCR技术结合基因组扫描,成功筛选出7株携带APD-CLDs基因簇的放线菌,并对这些菌株进行了基因组测序和分析。结果基因组分析结果表明,Micromonospora sp.FIM99-461、Micromonospora sp.FIM030746、Micromonospora sp.FIM060002、Micromonospora sp.FIM090374和Streptomyces sp.FIM18-1089菌株中存在与rakicidins生物合成基因簇,而Streptomyces sp.FIM15-0361和Streptomyces sp.FIM16-0599菌株的APD-CLDs基因簇则表现出一定的差异。模式菌株基因组服务器(TYGS)分析揭示,FIM16-0599、FIM18-1089和FIM99-461等3株为放线菌潜在新种。此外,在菌株FIM99-461、FIM03-0746、FIM06-0002和FIM09-0374菌株的发酵液中检测到了rakicidin A。结论建立了高效的APD-CLDs基因筛选策略,获得了多株APD-CLDs基因簇携带菌株。研究为APD-CLDs的生物合成研究和新药开发提供了新的资源,也为微生物次级代谢产物的筛选提供了新的方法和思路。
Objective To establish a genetic screening strategy for strains carrying the amino pentadienyl cyclic lipopeptides(APD-CLDs)gene cluster,and to screen and analyze such strains from an actinomycete strain library constructed in the laboratory.Methods By employing PCR technology and genome scanning,seven actinomycete strains carrying the APD-CLDs gene cluster were successfully screened,followed by genome sequencing and analysis.Results Genome analysis revealed that strains Micromonospora sp.FIM99-461,Micromonospora sp.FIM03-0746,Micromonospora sp.FIM06-0002,Micromonospora sp.FIM09-0374,and Streptomyces sp.FIM18-1089 contained gene clusters similar to those involved in the biosynthesis of rakicidins,whereas the APD-CLDs gene clusters in strains Streptomyces sp.FIM15-0361 and Streptomyces sp.FIM16-0599 showed some differences.The type strain genome server(TYGS)analysis identified FIM16-0599,FIM18-1089,and FIM99-461 as potential novel species of actinomycetes.Additionally,rakicidin A was detected in the fermentation broths of FIM99-461,FIM03-0746,FIM06-0002,and FIM09-0374.Conclusion This study established an efficient APD-CLDs gene screening strategy and successfully identified several strains carrying the APD-CLDs gene cluster.The findings provided new resources for the study of APD-CLDs biosynthesis and drug development,as well as new methods and insights for screening microbial secondary metabolites.
作者
方志锴
陈丽
林鹏辉
陈楠楠
江红
周剑
Fang Zhikai;Chen Li;Lin Penghui;Chen Nannan;Jiang Hong;Zhou Jian(Fujian Key Laboratory of Screening for Novel Microbial Products,Fujian Institute of Microbiology,Fuzhou 350007;School of Pharmacy,Fujian Medical University,Fuzhou 350122)
出处
《中国抗生素杂志》
北大核心
2025年第12期1444-1450,共7页
Chinese Journal of Antibiotics
基金
福建省公益类科研院所专项(No.2020R10050011)
福建省自然科学基金(No.2022J01387)。
关键词
氨基戊二烯环脂肽类化合物
生物合成基因簇
PCR筛选
基因组扫描
Amidopentadienoate-containing cyclolipodepsipeptides
Biosynthetic gene clusters
PCR screening
Genomic scanning
