Detection of the dipicolinic acid biomarker in Bacillus spores using Curie-point pyrolysis mass spectrometry and fourier transform infrared spectroscopy

被引:246
作者
Goodacre, R [1 ]
Shann, B
Gilbert, RJ
Timmins, ÉM
McGovern, AC
Alsberg, BK
Kell, DB
Logan, NA
机构
[1] Univ Wales, Inst Biol Sci, Aberystwyth SY23 3DD, Ceredigion, Wales
[2] Univ Wales, Dept Comp Sci, Aberystwyth SY23 3DB, Ceredigion, Wales
[3] Glasgow Caledonian Univ, Sch Biol & Biomed Sci, Glasgow G4 0BA, Lanark, Scotland
关键词
D O I
10.1021/ac990661i
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Thirty-six strains of aerobic endospore-forming bacteria confirmed by polyphasic taxonomic methods to belong to Bacillus amyloliquefaciens, Bacillus cereus, Bacillus licheniformis, Bacillus megaterium, Bacillus subtilis (including Bacillus niger and Bacillus globigii), Bacillus sphaericus,and Brevi laterosporus were grown axenically on nutrient agar, and vegetative and sporulated biomasses were analyzed by Curie-point pyrolysis mass spectrometry (PyMS) and diffuse reflectance-absorbance Fourier-transform infrared spectroscopy (FT-IK), Chemometric methods based on rule induction and genetic programming were used to determine the physiological state (vegetative cells or spores) correctly, and these methods produced mathematical rules which could be simply interpreted in biochemical terms. For PyMS it was found that m/z 105 was characteristic and is a pyridine ketoniun ion (C6H3ON+) obtained from the pyrolysis of dipicolinic acid (pyridine-2,6-dicarboxylic acid; DPA), a substance found in spores but not in vegetative cells; this was confirmed using pyrolysis-gas chromatography/mass spectrometry, In addition, a pyridine ring vibration at 1447-1439 cm(-1) from DPA was found to be highly characteristic of spores in FT-IR analysis. Thus, although the original data sets recorded hundreds of spectral variables from whole cells simultaneously, a simple biomarker can be used for the rapid and unequivocal detection of spores of these organisms.
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页码:119 / 127
页数:9
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