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  • 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
ZUSAMMENFASSUNG

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.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Kaliumphosphat, ACS reagent, ≥99.0%
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Kaliumphosphat, ACS reagent, ≥98%
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D-(+)-Glukose, ≥99.5% (GC)
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1-Butanol, 99.9%
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D-(+)-Glukose, powder, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.5%
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2-Butanon, ACS reagent, ≥99.0%
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1-Butanol, ACS reagent, ≥99.4%
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Natriumchlorid, DNase, RNase, and protease, none detected, ≥99% (titration)
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Kaliumphosphat, powder, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99.0%
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2-Butanon, suitable for HPLC, ≥99.7%
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Natriumchlorid -Lösung, 0.9% in water, BioXtra, suitable for cell culture
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1-Butanol, suitable for HPLC, ≥99.7%
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Natriumchlorid, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
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Dextrose, 97.5-102.0% anhydrous basis, meets EP, BP, JP, USP testing specifications
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Natriumchlorid -Lösung, 5 M in H2O, BioReagent, suitable for cell culture
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Ammoniumchlorid, ReagentPlus®, ≥99.5%
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Kaliumphosphat, buffer substance, anhydrous, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., 99.5-100.5%
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Ammoniumchlorid, suitable for cell culture, ≥99.5%
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D-(+)-Glukose, ≥99.5% (GC), BioXtra
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Kaliumphosphat, puriss. p.a., ACS reagent, anhydrous, ≥99.0% (T)
SAFC
Natriumchlorid -Lösung, 5 M
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Kaliumphosphat, meets analytical specification of Ph. Eur., NF, E340, anhydrous, 98-100.5% (calc. to the dried substance)
USP
Dextrose, United States Pharmacopeia (USP) Reference Standard
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1-Butanol, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
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Kaliumphosphat, ≥98.0%
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Kaliumphosphat, anhydrous, BioUltra, ≥99.0% (T)
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D-(+)-Glukose, BioUltra, anhydrous, ≥99.5% (sum of enantiomers, HPLC)
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Natriumchlorid -Lösung, BioUltra, ~5 M in H2O
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Indol, ≥99%
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Natriumchlorid, BioUltra, ≥99.5% (AT)