Extensive Turnover of Compatible Solutes in Cyanobacteria Revealed by Deuterium Oxide (D2O) Stable Isotope Probing

Richard Baran, Rebecca Lau, Benjamin P. Bowen, Spencer Diamond, Nick Jose, Ferran Garcia-Pichel, Trent R. Northen

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Cyanobacteria are important primary producers of organic matter in diverse environments on a global scale. While mechanisms of CO2 fixation are well understood, the distribution of the flow of fixed organic carbon within individual cells and complex microbial communities is less well characterized. To obtain a general overview of metabolism, we describe the use of deuterium oxide (D2O) to measure deuterium incorporation into the intracellular metabolites of two physiologically diverse cyanobacteria: a terrestrial filamentous strain (Microcoleus vaginatus PCC 9802) and a euryhaline unicellular strain (Synechococcus sp. PCC 7002). D2O was added to the growth medium during different phases of the diel cycle. Incorporation of deuterium into metabolites at nonlabile positions, an indicator of metabolite turnover, was assessed using liquid chromatography mass spectrometry. Expectedly, large differences in turnover among metabolites were observed. Some metabolites, such as fatty acids, did not show significant turnover over 12-24 h time periods but did turn over during longer time periods. Unexpectedly, metabolites commonly regarded to act as compatible solutes, including glutamate, glucosylglycerol, and a dihexose, showed extensive turnover compared to most other metabolites already after 12 h, but only during the light phase in the cycle. The observed extensive turnover is surprising considering the conventional view on compatible solutes as biosynthetic end points given the relatively slow growth and constant osmotic conditions. This suggests the possibility of a metabolic sink for some compatible solutes (e.g., into glycogen) that allows for rapid modulation of intracellular osmolarity. To investigate this, uniformly 13C-labeled Synechococcus sp. PCC 7002 were exposed to 12C glucosylglycerol. Following metabolite extraction, amylase treatment of methanol-insoluble polymers revealed 12C labeling of glycogen. Overall, our work shows that D2O probing is a powerful method for analysis of cyanobacterial metabolism including discovery of novel metabolic processes.

Original languageEnglish (US)
Pages (from-to)674-681
Number of pages8
JournalACS Chemical Biology
Volume12
Issue number3
DOIs
StatePublished - Mar 17 2017

Fingerprint

Deuterium Oxide
Cyanobacteria
Metabolites
Isotopes
Synechococcus
Deuterium
Glycogen
Amylases
Growth
Liquid Chromatography
Osmolar Concentration
Methanol
Glutamic Acid
Mass Spectrometry
Polymers
Fatty Acids
Carbon
Metabolism
Light
Liquid chromatography

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

Cite this

Extensive Turnover of Compatible Solutes in Cyanobacteria Revealed by Deuterium Oxide (D2O) Stable Isotope Probing. / Baran, Richard; Lau, Rebecca; Bowen, Benjamin P.; Diamond, Spencer; Jose, Nick; Garcia-Pichel, Ferran; Northen, Trent R.

In: ACS Chemical Biology, Vol. 12, No. 3, 17.03.2017, p. 674-681.

Research output: Contribution to journalArticle

Baran, Richard ; Lau, Rebecca ; Bowen, Benjamin P. ; Diamond, Spencer ; Jose, Nick ; Garcia-Pichel, Ferran ; Northen, Trent R. / Extensive Turnover of Compatible Solutes in Cyanobacteria Revealed by Deuterium Oxide (D2O) Stable Isotope Probing. In: ACS Chemical Biology. 2017 ; Vol. 12, No. 3. pp. 674-681.
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