Skip to Content
Merck
  • Investigation of volatile metabolites during growth of Escherichia coli and Pseudomonas aeruginosa by needle trap-GC-MS.

Investigation of volatile metabolites during growth of Escherichia coli and Pseudomonas aeruginosa by needle trap-GC-MS.

Analytical and bioanalytical chemistry (2014-08-26)
C Zscheppank, H L Wiegand, C Lenzen, J Wingender, U Telgheder
ABSTRACT

A new method for the growth-dependent headspace analysis of bacterial cultures by needle trap (NT)-gas chromatography-mass spectrometry (GC-MS) was established. NTs were used for the first time as enrichment technique for volatile organic compounds (VOCs) in the headspace of laboratory cultures. Reference strains of Escherichia coli and Pseudomonas aeruginosa were grown in different liquid culture media for 48 h at 36 °C. In the course of growth, bacterial culture headspace was analysed by NT-GC-MS. In parallel, the abiotic release of volatile organic compounds (VOC) from nutrient media was investigated by the same method. By examination of microbial headspace samples in comparison with those of uninoculated media, it could be clearly differentiated between products and compounds which serve as substrates. Specific microbial metabolites were detected and quantified during the stationary growth phase. P. aeruginosa produced dimethyl sulfide (max. 125 μg L(-1) < limits of quantification (LOQ)), 1-undecene (max. 164 μg L(-1)) and 2-nonanone (max. 200 μg L(-1)), whereas E. coli produced carbon disulfide, butanal and indole (max. 149 mg L(-1)). Both organisms produced isoprene.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Dimethyl sulfide, natural, ≥99%, FCC, FG
Sigma-Aldrich
Dimethyl sulfide, ≥99%, FCC, FG
Supelco
Dextrose, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Ammonium-14N chloride, 99.99 atom % 14N, 15N-depleted, 99% (CP)
Sigma-Aldrich
2-Butanone, FCC, FG
Sigma-Aldrich
Potassium phosphate dibasic solution, 1.0 M
Sigma-Aldrich
Potassium phosphate monobasic, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
Sigma-Aldrich
Dextrose, 97.5-102.0% anhydrous basis, meets EP, BP, JP, USP testing specifications
Sigma-Aldrich
Butyl alcohol, natural, ≥99.5%, FCC, FG
Sigma-Aldrich
Dimethyl sulfide, anhydrous, ≥99.0%
Sigma-Aldrich
Dimethyl disulfide, ≥99.0%
Supelco
Dimethyl sulfide, analytical standard
USP
Dextrose, United States Pharmacopeia (USP) Reference Standard
USP
Monobasic potassium phosphate, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Potassium phosphate monobasic, ACS reagent, ≥99.0%
Sigma-Aldrich
Potassium phosphate dibasic, ACS reagent, ≥98%
Sigma-Aldrich
2-Butanone, ACS reagent, ≥99.0%
Supelco
Methyl Ethyl Ketone, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
2-Butanone, suitable for HPLC, ≥99.7%
Sigma-Aldrich
2-Methylbutyraldehyde, ≥95%, FG
Sigma-Aldrich
Sodium chloride, BioUltra, Molecular Biology, ≥99.5% (AT)
Sigma-Aldrich
Sodium chloride solution, BioUltra, Molecular Biology, ~5 M in H2O
Supelco
1-Butanol, analytical standard
Sigma-Aldrich
D-(+)-Glucose, BioUltra, anhydrous, ≥99.5% (sum of enantiomers, HPLC)
Sigma-Aldrich
Indole, ≥99%, FG
Sigma-Aldrich
1-Butanol, anhydrous, 99.8%
Sigma-Aldrich
D-Glucose-12C6, 16O6, 99.9 atom % 16O, 99.9 atom % 12C
Sigma-Aldrich
D-(+)-Glucose, ACS reagent
Sigma-Aldrich
D-(+)-Glucose, ≥99.5% (GC)
Sigma-Aldrich
Sodium chloride, BioXtra, ≥99.5% (AT)