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Sensitivity to the two peptide bacteriocin plantaricin EF is dependent on CorC a membrane‐bound magnesium/cobalt efflux protein

机译:对两种肽细菌素植物霉素EF的敏感性取决于膜结合的镁/钴外排蛋白CorC

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摘要

Lactic acid bacteria produce a variety of antimicrobial peptides known as bacteriocins. Most bacteriocins are understood to kill sensitive bacteria through receptor‐mediated disruptions. Here, we report on the identification of the plantaricin EF (PlnEF) receptor. Spontaneous PlnEF‐resistant mutants of the PlnEF‐indicator strain NCIMB 700965 (LP965) were isolated and confirmed to maintain cellular ATP levels in the presence of PlnEF. Genome comparisons resulted in the identification of a single mutated gene annotated as the membrane‐bound, magnesium/cobalt efflux protein CorC. All isolates contained a valine (V) at position 334 instead of a glycine (G) in a cysteine‐β‐synthase domain at the C‐terminal region of CorC. In silico template‐based modeling of this domain indicated that the mutation resides in a loop between two β‐strands. The relationship between PlnEF, CorC, and metal homeostasis was supported by the finding that PlnEF‐resistance was lost when PlnEF was applied together with high concentrations of Mg , Co , Zn , or Cu . Lastly, PlnEF sensitivity was increased upon heterologous expression of LP965 but not the G334V CorC mutant in the PlnEF‐resistant strain BL23. These results show that PlnEF kills sensitive bacteria by targeting CorC.
机译:乳酸菌可产生多种抗菌肽,称为细菌素。大多数细菌素被理解为通过受体介导的破坏杀死敏感细菌。在这里,我们报告了对植物抑素EF(PlnEF)受体的鉴定。分离出PlnEF指示剂菌株NCIMB 700965(LP965)的自发性PlnEF耐药突变体,并确认在存在PlnEF的情况下可以维持细胞的ATP水平。通过基因组比较,鉴定出一个注释为膜结合的镁/钴外排蛋白CorC的单个突变基因。所有分离株在CorC的C端区域的半胱氨酸-β-合酶结构域中的334位均含有缬氨酸(V)而不是甘氨酸(G)。在基于计算机键盘模板的此域建模中,该突变位于两个β链之间的环中。发现当将PlnEF与高浓度的Mg,Co,Zn或Cu一起使用时会失去PlnEF电阻,这支持了PlnEF,CorC和金属稳态之间的关系。最后,在耐PlnEF的菌株BL23中,LP965的异源表达增加了PlnEF的敏感性,但没有增加G334V CorC突变体的敏感性。这些结果表明,PlnEF通过靶向CorC杀死敏感细菌。

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