Article

Longevity of major coenzymes allows minimal de novo synthesis in microorganisms

  • Nature Microbiology 2, Article number: 17073 (2017)
  • doi:10.1038/nmicrobiol.2017.73
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Abstract

Coenzymes are vital for cellular metabolism and act on the full spectrum of enzymatic reactions. Intrinsic chemical reactivity, enzyme promiscuity and high flux through their catalytic cycles make coenzymes prone to damage. To counteract such compromising factors and ensure stable levels of functional coenzymes, cells use a complex interplay between de novo synthesis, salvage, repair and degradation. However, the relative contribution of these factors is currently unknown, as is the overall stability of coenzymes in the cell. Here, we use dynamic 13C-labelling experiments to determine the half-life of major coenzymes of Escherichia coli. We find that coenzymes such as pyridoxal 5-phosphate, flavins, nicotinamide adenine dinucleotide (phosphate) and coenzyme A are remarkably stable in vivo and allow biosynthesis close to the minimal necessary rate. In consequence, they are essentially produced to compensate for dilution by growth and passed on over generations of cells. Exceptions are antioxidants, which are short-lived, suggesting an inherent requirement for increased renewal. Although the growth-driven turnover of stable coenzymes is apparently subject to highly efficient end-product homeostasis, we exemplify that coenzyme pools are propagated in excess in relation to actual growth requirements. Additional testing of Bacillus subtilis and Saccharomyces cerevisiae suggests that coenzyme longevity is a conserved feature in biology.

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Acknowledgements

The authors thank U. Sauer and J.-C. Portais for discussions and comments on the manuscript and P. Christen for technical support with LC-MS. This work was supported by the Swiss National Science Foundation (grant no. 31003A-173094) and ETH Zurich.

Author information

Affiliations

  1. Institute of Microbiology, ETH Zurich, 8093 Zurich, Switzerland

    • Johannes Hartl
    • , Patrick Kiefer
    • , Fabian Meyer
    •  & Julia A. Vorholt

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Contributions

J.H., P.K., F.M. and J.A.V. planned the project and designed the experiments. J.H. and F.M. performed the experiments. J.H. and P.K. set-up LC-MS methods and data analysis. J.H. performed measurements and data analysis. J.H. and J.A.V. wrote the manuscript with input from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Julia A. Vorholt.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Figures 1-9, Supplementary Tables 1-6, Supplementary Methods, Supplementary References.

Excel files

  1. 1.

    Supplementary Data 1

    Excel file containing analysed LC-MS and LC-MS/MS data from a long-term dynamic labelling switch experiment (12C to 13C) with Escherichia coli.

  2. 2.

    Supplementary Data 2

    Excel file containing analysed LC-MS and LC-MS/MS data from a long-term dynamic labelling switch experiment (13C to 12C) with Escherichia coli.

  3. 3.

    Supplementary Data 3

    Excel file containing analysed LC-MS and LC-MS/MS data from a long-term dynamic labelling switch experiment (12C to 13C) with Escherichia coli Δfdx.

  4. 4.

    Supplementary Data 4

    Excel file containing analysed LC-MS and LC-MS/MS data from a long-term dynamic labelling switch experiment (12C to 13C) with Escherichia coli.

  5. 5.

    Supplementary Data 5

    Excel file containing analysed LC-MS and LC-MS/MS data from a long-term dynamic labelling switch experiment (12C to 13C) with Saccharomyces cerevisiae.