Mirkovic Nemanja, Polovic Natalija, Vukotic Goran, Jovcic Branko, Miljkovic Marija, Radulovic Zorica, Diep Dzung B, Kojic Milan
Laboratory for Molecular Microbiology, Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
Department for Food Microbiology, Faculty of Agriculture, University of Belgrade, Belgrade, Serbia.
Appl Environ Microbiol. 2016 Apr 4;82(8):2555-2562. doi: 10.1128/AEM.03988-15. Print 2016 Apr.
Bacteriocin producers normally possess dedicated immunity systems to protect themselves from their own bacteriocins.Lactococcus lactis strains LMG2081 and BGBM50 are known as lactococcin G producers. However, BGBM50 was sensitive to LMG2081, which indicated that LMG2081 might produce additional bacteriocins that are not present in BGBM50. Therefore, whole-genome sequencing of the two strains was performed, and a lantibiotic operon (called lctLMG) was identified in LMG2081 but not in BGBM50. The lctLMG operon contains six open reading frames; the first three genes,lmgA ,lmgM, and lmgT, are involved in the biosynthesis and export of bacteriocin, while the other three genes,lmgF,lmgE, and lmgG, are involved in lantibiotic immunity. Mutational analysis confirmed that the lctLMG operon is responsible for the additional antimicrobial activity. Specifically, site-directed mutation within this operon rendered LMG2081 inactive toward BGBM50. Subsequent purification and electrospray ionization-time of flight mass spectrometric analysis confirmed that the lantibiotic bacteriocin called lacticin LMG is exported as a 25-amino-acid peptide. Lacticin LMG is highly similar to the lacticin 481 group. It is interesting that a bacteriocin producer produces two different classes of bacteriocins, whose operons are located in the chromosome and a plasmid.
细菌素产生菌通常拥有专门的免疫系统来保护自身免受自身产生的细菌素的影响。乳酸乳球菌菌株LMG2081和BGBM50是已知的乳球菌素G产生菌。然而,BGBM50对LMG2081敏感,这表明LMG2081可能产生了BGBM50中不存在的其他细菌素。因此,对这两个菌株进行了全基因组测序,在LMG2081中鉴定出一个羊毛硫抗生素操纵子(称为lctLMG),而在BGBM50中未鉴定到。lctLMG操纵子包含六个开放阅读框;前三个基因lmgA、lmgM和lmgT参与细菌素的生物合成和输出,而其他三个基因lmgF、lmgE和lmgG参与羊毛硫抗生素免疫。突变分析证实lctLMG操纵子负责额外的抗菌活性。具体而言,该操纵子内的定点突变使LMG2081对BGBM50失去活性。随后的纯化和电喷雾电离-飞行时间质谱分析证实,名为乳酸乳球菌素LMG的羊毛硫抗生素细菌素以25个氨基酸的肽形式输出。乳酸乳球菌素LMG与乳酸乳球菌素481组高度相似。有趣的是,一种细菌素产生菌产生了两类不同的细菌素,其操纵子分别位于染色体和质粒上。