Université d'Avignon et des Pays de Vaucluse, UMR408, Sécurité et Qualité des Produits d'Origine Végétale, F-84000 Avignon, France.
J Bacteriol. 2011 Apr;193(7):1757-66. doi: 10.1128/JB.00024-11. Epub 2011 Feb 4.
The diarrheal potential of a Bacillus cereus strain is essentially dictated by the amount of secreted nonhemolytic enterotoxin (Nhe). Expression of genes encoding Nhe is regulated by several factors, including the metabolic state of the cells. To identify metabolic sensors that could promote communication between central metabolism and nhe expression, we compared four strains of the B. cereus group in terms of metabolic and nhe expression capacities. We performed growth performance measurements, metabolite analysis, and mRNA measurements of strains F4430/73, F4810/72, F837/76, and PA cultured under anoxic and fully oxic conditions. The results showed that expression levels of nhe and ldhA, which encodes lactate dehydrogenase A (LdhA), were correlated in both aerobically and anaerobically grown cells. We examined the role of LdhA in the F4430/73 strain by constructing an ldhA mutant. The ldhA mutation was more deleterious to anaerobically grown cells than to aerobically grown cells, causing growth limitation and strong deregulation of key fermentative genes. More importantly, the ldhA mutation downregulated enterotoxin gene expression under both anaerobiosis and aerobiosis, with a more pronounced effect under anaerobiosis. Therefore, LdhA was found to exert a major control on both fermentative growth and enterotoxin expression, and it is concluded that there is a direct link between fermentative metabolism and virulence in B. cereus. The data presented also provide evidence that LdhA-dependent regulation of enterotoxin gene expression is oxygen independent. This study is the first report to describe a role of a fermentative enzyme in virulence in B. cereus.
一种蜡样芽胞杆菌菌株的腹泻潜力主要取决于分泌的非溶血肠毒素 (Nhe) 的量。编码 Nhe 的基因的表达受多种因素的调节,包括细胞的代谢状态。为了确定可以促进中心代谢和 nhe 表达之间通讯的代谢传感器,我们比较了 B. cereus 组的四个菌株在代谢和 nhe 表达能力方面的差异。我们进行了生长性能测量、代谢物分析和 F4430/73、F4810/72、F837/76 和 PA 菌株在缺氧和完全有氧条件下的 mRNA 测量。结果表明,在好氧和厌氧生长的细胞中,nhe 和编码乳酸脱氢酶 A (LdhA) 的 ldhA 的表达水平相关。我们通过构建 ldhA 突变体来研究 LdhA 在 F4430/73 菌株中的作用。ldhA 突变对厌氧生长的细胞比好氧生长的细胞更具危害性,导致生长受限和关键发酵基因的强烈失调。更重要的是,ldhA 突变在厌氧和需氧条件下均下调肠毒素基因表达,在厌氧条件下的影响更为明显。因此,LdhA 对发酵生长和肠毒素表达均具有主要的控制作用,并且可以得出结论,在蜡样芽胞杆菌中,发酵代谢和毒力之间存在直接联系。所提供的数据还提供了证据表明,LdhA 依赖性的肠毒素基因表达调控与氧无关。这项研究首次报道了一种发酵酶在蜡样芽胞杆菌毒力中的作用。