Department of Developmental and Surgical Sciences, University of Minnesota, Minneapolis, MN 55455, USA.
Department of Earth and Environmental Sciences, University of Minnesota, Minneapolis, MN 55455, USA.
Lett Appl Microbiol. 2023 Feb 16;76(2). doi: 10.1093/lambio/ovad017.
The uptake and storage of extracellular orthophosphate (Pi) by polyphosphate (polyP) accumulating bacteria may contribute to mineral dissolution in the oral cavity. To test the effect of potential inhibitors of polyP kinases on Rothia dentocariosa, gallein (0, 25, 50, and 100 μM) and fluoride (0, 50, and 100 ppm) were added to R. dentocariosa cultures grown in brain-heart infusion broth. At a late log growth phase (8 h), extracellular Pi was measured using an ascorbic acid assay, and polyP was isolated from bacterial cells treated with RNA/DNAases using a neutral phenol/chloroform extraction. Extracts were hydrolyzed and quantified as above. Gallein and fluoride had minor effects on bacterial growth with NaF having a direct effect on media pH. Gallein (≥25 μM) and fluoride (≥50 ppm) attenuated the bacterial drawdown of extracellular Pi by 56.7% (P < 0.05) and 37.3% (P < 0.01). There was a corresponding polyP synthesis decrease of 73.2% (P < 0.0001) from gallein and 83.1% (P < 0.0001) from fluoride. Attenuated total reflectance-Fourier-transform infrared spectroscopy validated the presence of polyP and its reduced concentration in R. dentocariosa bacterial cells following gallein and fluoride treatment. Rothia dentocariosa can directly change extracellular Pi and accumulate intracellular polyP, but the mechanism is attenuated by gallein and NaF.
聚磷酸盐(polyP)积累菌对细胞外正磷酸盐(Pi)的摄取和储存可能有助于口腔内矿物质的溶解。为了测试聚磷酸盐激酶潜在抑制剂对 Rothia dentocariosa 的影响,将 gallein(0、25、50 和 100μM)和氟化物(0、50 和 100ppm)添加到在脑心浸液肉汤中生长的 R.dentocariosa 培养物中。在对数生长期晚期(8h),使用抗坏血酸测定法测量细胞外 Pi,并用 RNA/DNA 酶处理细菌细胞,用中性苯酚/氯仿萃取法分离 polyP。如上所述水解并定量提取物。gallein 和氟化物对细菌生长的影响较小,而 NaF 对培养基 pH 有直接影响。gallein(≥25μM)和氟化物(≥50ppm)分别降低了 56.7%(P<0.05)和 37.3%(P<0.01)的细菌细胞外 Pi 消耗。相应的 polyP 合成分别降低了 73.2%(P<0.0001)来自 gallein 和 83.1%(P<0.0001)来自氟化物。衰减全反射-傅里叶变换红外光谱验证了 gallein 和氟化物处理后 Rothia dentocariosa 细菌细胞中 polyP 的存在及其浓度降低。Rothia dentocariosa 可以直接改变细胞外 Pi 并积累细胞内 polyP,但 gallein 和 NaF 会减弱这种机制。