Institute for Science and Technology in Medicine, School of Medicine, Keele University, Thornburrow Drive, Hartshill, Stoke-on-Trent, ST4 7QB, UK.
Rapid Commun Mass Spectrom. 2011 Aug 15;25(15):2163-72. doi: 10.1002/rcm.5099.
Preliminary measurements have been made of the volatile compounds emitted by the bacterium E. coli JM109 cultured in the commonly used media Dulbecco's modified Eagle's medium (DMEM) and lysogeny broth (LB) using selected ion flow tube mass spectrometry, SIFT-MS, as a step towards the real time, non-invasive monitoring of accidental infections of mammalian cell cultures. In one procedure, the culture medium alone and the E. coli cells/medium combination were held at 37 °C in bottles sealed with septa for a given time period, usually overnight, to allow the bacterium to proliferate, after which the captured headspace was analysed directly by SIFT-MS. Several compounds were seen to be produced by the E. coli cells that depended on the liquid medium used: when cultured in DMEM, copious amounts of ethanol, acetaldehyde and hydrogen sulphide were produced; in LB ammonia is the major volatile product. In a second procedure, to ensure aerobic conditions prevailed in the cell culture, selected volatile compounds were monitored by SIFT-MS in real time for several hours above the open-to-air E. coli/DMEM culture held at close to 37 °C. The temporal variations in the concentrations of some compounds, which reflect their production rates in the culture, indicate maxima. Thus, the maxima in the ethanol and acetaldehyde production are a reflection of the reduction of glucose from the DMEM by the vigorous E. coli cells and the maximum in the hydrogen sulphide level is an indication of the loss of the sulphur-bearing amino acids from the DMEM. Serendipitously, emissions from DMEM inadvertently infected with the bacterium C. testosteroni were observed when large quantities of ammonia were seen to be produced. The results of this preliminary study suggest that monitoring volatile compounds might assist in the early detection of bacterial infection in large-scale bioreactors.
初步测量已经进行了细菌大肠杆菌 JM109 发出的挥发性化合物培养在常用的介质 Dulbecco 修改后的 Eagle 氏培养基 (DMEM) 和溶菌肉汤 (LB) 使用选择离子流管质谱,SIFT-MS,作为一个实时,非侵入性监测哺乳动物细胞培养的偶然感染的方法。在一个程序中,单独的培养基和大肠杆菌细胞/培养基组合在密封瓶中保持在 37°C 一段时间,通常是过夜,以允许细菌增殖,之后直接通过 SIFT-MS 分析捕获的顶空。发现几种由大肠杆菌细胞产生的化合物依赖于所用的液体培养基:在 DMEM 中培养时,会产生大量的乙醇、乙醛和硫化氢;在 LB 中,氨气是主要的挥发性产物。在第二个程序中,为了确保细胞培养中存在有氧条件,通过 SIFT-MS 实时监测几种挥发性化合物数小时,这些化合物在接近 37°C 的空气中开放的大肠杆菌/LB 培养基上方。一些化合物浓度的时间变化反映了它们在培养物中的产生速率,表明有最大值。因此,乙醇和乙醛产生的最大值反映了 DMEM 中葡萄糖被活力旺盛的大肠杆菌细胞还原,以及硫化氢水平的最大值表明 DMEM 中含硫氨基酸的损失。偶然地,当观察到大量氨产生时,无意中被细菌 C. testosteroni 感染的 DMEM 的排放被观察到。这项初步研究的结果表明,监测挥发性化合物可能有助于在大规模生物反应器中早期检测细菌感染。