Detection of cyclic lipopeptide biomarkers from Bacillus species using atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry.
Detection of cyclic lipopeptide biomarkers from Bacillus species using atmospheric pressure matrix-assisted laser desorption/ionization mass spectrometry.
复制标题
使用大气压基质辅助激光解吸/电离质谱法检测芽孢杆菌属物种的环状脂肽生物标志物。
DOI：
10.1021/ac020506v
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发表时间：
2003
影响因子：
7.4
通讯作者：
Doroshenko,VladimirM
中科院分区：
文献类型：
--
作者：
Madonna,AngeloJ;Voorhees,KentJ;Taranenko,NelliI;Laiko,VictorV;Doroshenko,VladimirM
介绍了一种新的微生物检测方法，该方法使用大气压基质辅助激光解吸/电离与离子阱质谱仪分析全细胞细菌。这种新的方法进行了测试与冻干孢子和培养物的芽孢杆菌球（BG）在琼脂培养基上生长4天或更长的时间。在生长的每个阶段，发现生物标志物（鉴定为称为芬枯草菌素和表面活性素的环脂肽）可通过脉冲紫外激光照射与α-氰基-4-羟基肉桂酸共结晶的完整BG细胞（约1.5 mg）进行检测。此外，通过对环脂肽的前体离子进行串联质谱，获得了确定的氨基酸序列信息。扩大调查范围，包括检查从大气中收集并直接沉积在双面胶带上的气溶胶化BG孢子。随后对回收的孢子进行分析，结果产生了与芬枯草菌素一致的质量峰。对芽孢杆菌属物种进行了分析比较，并显示质谱峰也被鉴定为源自各种环脂肽。使用脉冲红外激光器作为激发源进行进一步的研究，以分析悬浮在0.03 M柠檬酸铵和甘油的基质中的BG细胞（约1.5 mg），导致产生丰霉素和表面活性素的特征性离子。
A novel approach to microbial detection using atmospheric pressure matrix-assisted laser desorption/ionization with an ion trap mass spectrometer to analyze whole cell bacteria is introduced. This new approach was tested with lyophilized spores and cultures ofBacillusglobigii(BG) grown on agar media for 4 days or longer. At each stage of growth, it was found that biomarkers, identified as cyclic lipopeptides known as fengycin and surfactin, could be detected by pulsed ultraviolet laser irradiation of intact BG cells (∼5 mg) cocrystallized with α-cyano-4-hydroxycinnamic acid. Furthermore, definitive amino acid sequence information was obtained by performing tandem mass spectrometry on the precursor ions of the cyclic lipopeptides. The investigation was broadened to include the examination of aerosolized BG spores collected from the atmosphere and directly deposited onto double-sided tape. Subsequent analysis of the recovered spores resulted in the production of mass peaks consistent with fengycin. OtherBacillusspecies were analyzed for comparison and showed mass spectral peaks also identified as originating from various cyclic lipopeptides. Further studies were conducted using a pulsed infrared laser as the excitation source to analyze BG cells (∼5 mg) suspended in a matrix of 0.03 M ammonium citrate and glycerol resulting in the production of ions characteristic of fengycin and surfactin.