粪肠球菌CRL1879细菌素基因的基因组挖掘与转录分析

N. Suárez, J. Bonacina, E. Hebert, L. Saavedra
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引用次数: 8

摘要

从9种手工奶酪中分离的151株细菌中,屎肠球菌(Enterococcus faecium) CRL 1879对食源性单核增生李斯特菌具有抗菌活性。分离物在无细胞上清液中产生蛋白酶k敏感化合物。基因组分析显示存在肠球菌蛋白A、肠球菌蛋白B、肠球菌蛋白P、肠球菌蛋白se - k4样和肠球菌蛋白X生物合成基因簇。利用生物信息学工具检测了一种推定的双组分细菌素的核苷酸序列,这里将其命名为enterocin CRL1879αβ。利用实时荧光定量PCR (qRT-PCR)对所有细菌素基因进行转录分析,揭示了各肠霉素基因的转录水平不同。最后,对251株粪肠杆菌的细菌素基因分布进行了分析,并与粪肠杆菌CRL1879中的细菌素基因进行了比较。判别分析表明,细菌素基因在肠球菌中广泛分布,与菌株的来源无关。本文中提出的结果是一个独特的发现,因为这是第一次证明从手工奶酪中分离出的粪肠杆菌菌株具有完整的遗传机制,可以产生六种II类和一种III类细菌素。
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Genome Mining and Transcriptional Analysis of Bacteriocin Genes in Enterococcus faecium CRL1879
Among 151 bacterial isolates from nine artisanal cheeses, Enterococcus faecium CRL 1879 showed antibacterial activity against the food-borne pathogen Listeria monocytogenes. The isolate produced a proteinase K-sensitive compound in the cell free supernatant. Genome analysis demonstrated the presence of enterocin A, enterocin B, enterocin P, enterocin SE-K4-like and enterocin X biosynthetic gene clusters. Nucleotide sequences encoding for a putative two-component bacteriocin were detected using bioinformatics tools, here named enterocin CRL1879αβ. A transcriptional analysis of all bacteriocin genes by quantitative real time PCR analysis (qRT-PCR) revealed the transcription of each enterocin gene at different levels. Finally, analysis of bacteriocin genes distribution in 251 E. faecium bioprojects was performed and compared to those identify in E. faecium CRL1879. The discriminative analysis demonstrated that bacteriocin genes are widely distributed among Enterococcus, independently of the origin of the strain. The results presented in this paper represent a unique finding since this is the first demonstration of an E. faecium strain isolated from an artisanal cheese with the complete genetic machinery to produce six classes II and one class III bacteriocins.
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