Time-resolved selected ion flow tube mass spectrometric quantification of the volatile compounds generated by E. coli JM109 cultured in two different media

被引:34
作者
Chippendale, Thomas W. E. [2 ]
Spanel, Patrik [1 ,2 ]
Smith, David [2 ]
机构
[1] Acad Sci Czech Republ, J Heyrovsky Inst Phys Chem, CR-18223 Prague 8, Czech Republic
[2] Keele Univ, Sch Med, Inst Sci & Technol Med, Stoke On Trent ST4 7QB, Staffs, England
基金
英国工程与自然科学研究理事会;
关键词
SIFT-MS; IN-VITRO; PSEUDOMONAS-AERUGINOSA; ORGANIC-COMPOUNDS; HUMID AIR; BREATH; HEADSPACE; METABOLITES; ORIGIN; GROWTH;
D O I
10.1002/rcm.5099
中图分类号
Q5 [生物化学];
学科分类号
070307 [化学生物学];
摘要
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 degrees 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 degrees 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. Copyright (C) 2011 John Wiley & Sons, Ltd.
引用
收藏
页码:2163 / 2172
页数:10
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