Efficacy of bacterial bioremediation

Demonstration of complete incorporation of hydrocarbons into membrane phospholipids from Rhodococcus hydrocarbon degrading bacteria by electrospray ionization fourier transform ion cyclotron resonance mass spectrometry

Ryan P. Rodgers, Erin N. Blumer, Mark Emmett, Alan G. Marshall

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We present a method and example to establish complete incorporation of hydrocarbons into membrane phospholipids (and their constituent individual fatty acids and polar headgroup) of putatively bioremediative bacteria. Bacteria are grown on minimal media containing a specified carbon source (in this case, C16 and C18 alkanes), either natural abundance (99% 12C) or enriched (99% 13C). After extraction (but no other prior separation) of the membrane lipids, electrospray ionization yields a negative-ion FT-ICR mass spectrum containing prominent phospholipid parent ions. If 13C-enriched hydrocarbon incorporation is complete, then the mass of the parent ion will increase by n Da, in which n is the number of its constituent carbon atoms; moreover, the 13C isotopic distribution pattern will be reversed. The identities of the constituent fatty acids and polar headgroup are obtained by collisional dissociation (MS/MS), and their extent of 13C incorporation determined individually. The method is demonstrated for Rhodecoccus rhodochrous (ATCC 53968), for which all 44 carbons of a representative phosphatidylinositol are shown to derive from the hydrocarbon source. Interestingly, although only C16 and C18 alkanes are provided in the growth medium, the bacteria synthesize uniformly enriched C16:0 and C19:0 fatty acids.

Original languageEnglish (US)
Pages (from-to)535-540
Number of pages6
JournalEnvironmental Science and Technology
Volume34
Issue number3
DOIs
StatePublished - Feb 1 2000
Externally publishedYes

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Cyclotron resonance
Electrospray ionization
Bioremediation
Phospholipids
Hydrocarbons
phospholipid
Fatty acids
bioremediation
Fourier transform
Mass spectrometry
Bacteria
Fourier transforms
Alkanes
ionization
Demonstrations
Fatty Acids
fatty acid
Carbon
mass spectrometry
Ions

ASJC Scopus subject areas

  • Environmental Science(all)
  • Environmental Chemistry
  • Environmental Engineering

Cite this

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abstract = "We present a method and example to establish complete incorporation of hydrocarbons into membrane phospholipids (and their constituent individual fatty acids and polar headgroup) of putatively bioremediative bacteria. Bacteria are grown on minimal media containing a specified carbon source (in this case, C16 and C18 alkanes), either natural abundance (99{\%} 12C) or enriched (99{\%} 13C). After extraction (but no other prior separation) of the membrane lipids, electrospray ionization yields a negative-ion FT-ICR mass spectrum containing prominent phospholipid parent ions. If 13C-enriched hydrocarbon incorporation is complete, then the mass of the parent ion will increase by n Da, in which n is the number of its constituent carbon atoms; moreover, the 13C isotopic distribution pattern will be reversed. The identities of the constituent fatty acids and polar headgroup are obtained by collisional dissociation (MS/MS), and their extent of 13C incorporation determined individually. The method is demonstrated for Rhodecoccus rhodochrous (ATCC 53968), for which all 44 carbons of a representative phosphatidylinositol are shown to derive from the hydrocarbon source. Interestingly, although only C16 and C18 alkanes are provided in the growth medium, the bacteria synthesize uniformly enriched C16:0 and C19:0 fatty acids.",
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AB - We present a method and example to establish complete incorporation of hydrocarbons into membrane phospholipids (and their constituent individual fatty acids and polar headgroup) of putatively bioremediative bacteria. Bacteria are grown on minimal media containing a specified carbon source (in this case, C16 and C18 alkanes), either natural abundance (99% 12C) or enriched (99% 13C). After extraction (but no other prior separation) of the membrane lipids, electrospray ionization yields a negative-ion FT-ICR mass spectrum containing prominent phospholipid parent ions. If 13C-enriched hydrocarbon incorporation is complete, then the mass of the parent ion will increase by n Da, in which n is the number of its constituent carbon atoms; moreover, the 13C isotopic distribution pattern will be reversed. The identities of the constituent fatty acids and polar headgroup are obtained by collisional dissociation (MS/MS), and their extent of 13C incorporation determined individually. The method is demonstrated for Rhodecoccus rhodochrous (ATCC 53968), for which all 44 carbons of a representative phosphatidylinositol are shown to derive from the hydrocarbon source. Interestingly, although only C16 and C18 alkanes are provided in the growth medium, the bacteria synthesize uniformly enriched C16:0 and C19:0 fatty acids.

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