Dental Research Institute, University of Toronto, Toronto, Ontario, Canada.
J Bacteriol. 2010 Feb;192(4):984-93. doi: 10.1128/JB.01169-09. Epub 2009 Dec 18.
Glutamate contributes to the acid tolerance response (ATR) of many Gram-negative and Gram-positive bacteria, but its role in the ATR of the oral bacterium Streptococcus mutans is unknown. This study describes the discovery and characterization of a glutamate transporter operon designated glnQHMP (Smu.1519 to Smu.1522) and investigates its potential role in acid tolerance. Deletion of glnQHMP resulted in a 95% reduction in transport of radiolabeled glutamate compared to the wild-type UA159 strain. The addition of glutamate to metabolizing UA159 cells resulted in an increased production of acidic end products, whereas the glnQHMP mutant produced less lactic acid than UA159, suggesting a link between glutamate metabolism and acid production and possible acid tolerance. To investigate this possibility, we conducted a microarray analysis with glutamate and under pH 5.5 and pH 7.5 conditions which showed that expression of the glnQHMP operon was downregulated by both glutamate and mild acid. We also measured the growth kinetics of UA159 and its glnQHMP-negative derivative at pH 5.5 and found that the mutant doubled at a much slower rate than the parent strain but survived at pH 3.5 significantly better than the wild type. Taken together, these findings support the involvement of the glutamate transporter operon glnQHMP in the acid tolerance response in S. mutans.
谷氨酸有助于许多革兰氏阴性和革兰氏阳性细菌的耐酸反应(ATR),但其在口腔细菌变形链球菌的 ATR 中的作用尚不清楚。本研究描述了谷氨酸转运子操纵子 glnQHMP(Smu.1519 到 Smu.1522)的发现和特征,并研究了其在耐酸中的潜在作用。与野生型 UA159 菌株相比,glnQHMP 的缺失导致放射性标记的谷氨酸转运减少了 95%。将谷氨酸添加到代谢性 UA159 细胞中会导致酸性终产物的产量增加,而 glnQHMP 突变体产生的乳酸比 UA159 少,这表明谷氨酸代谢与产酸之间存在联系,并且可能与耐酸有关。为了研究这种可能性,我们进行了微阵列分析,发现谷氨酸和 pH 5.5 和 pH 7.5 条件下,glnQHMP 操纵子的表达均受谷氨酸和轻度酸化的下调。我们还测量了 UA159 及其 glnQHMP 阴性衍生物在 pH 5.5 下的生长动力学,发现突变体的倍增速度比亲本菌株慢得多,但在 pH 3.5 下的存活率明显高于野生型。综上所述,这些发现支持谷氨酸转运子操纵子 glnQHMP 参与变形链球菌的耐酸反应。