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Engineering and analysis of a Saccharomyces cerevisiae strain that uses formaldehyde as an auxiliary substrate

Baerends, R. J. S., de Hulster, E., Geertman, J-M. A., Daran, J-M., van Maris, A. J. A., Veenhuis, M., van der Klei, I. J. & Pronk, J. T., May-2008, In : Applied and environmental microbiology. 74, 10, p. 3182-3188 7 p.

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  • Richard J. S. Baerends
  • Erik de Hulster
  • Jan-Maarten A. Geertman
  • Jean-Marc Daran
  • Antonius J. A. van Maris
  • Marten Veenhuis
  • Ida J. van der Klei
  • Jack T. Pronk

We demonstrated that formaldehyde can be efficiently coutilized by an engineered Saccharomyces cerevisiae strain that expresses Hansenula polymorpha genes encoding formaldehyde dehydrogenase (FLD1) and formate dehydrogenase (FMD), in contrast to wild-type strains. Initial chemostat experiments showed that the engineered strain coutilized formaldehyde with glucose, but these mixed-substrate cultures failed to reach steadystate conditions and did not exhibit an increased biomass yield on glucose. Subsequent transcriptome analyses of chemostat cultures of the engineered strain, grown on glucose-formaldehyde mixtures, indicated that the presence of formaldehyde in the feed caused biotin limitations. Further transcriptome analysis demonstrated that this biotin inactivation was prevented by using separate formaldehyde and vitamin feeds. Using this approach, steady-state glucose-limited chemostat cultures were obtained that coutilized glucose and formaldehyde. Coutilization of formaldehyde under these conditions resulted in an enhanced biomass yield of the glucose-limited cultures. The biomass yield was quantitatively consistent with the use of formaldehyde as an auxiliary substrate that generates NADH and subsequently, via oxidative phosphorylation, ATP. On an electron pair basis, the biomass yield increase observed with formaldehyde was larger than that observed previously for formate, which is tentatively explained by different modes of formate and formaldehyde transport in S. cerevisiae.

Original languageEnglish
Pages (from-to)3182-3188
Number of pages7
JournalApplied and environmental microbiology
Volume74
Issue number10
Publication statusPublished - May-2008

    Keywords

  • LIMITED CHEMOSTAT CULTURES, ADDITIONAL ENERGY-SOURCE, MOLECULAR CHARACTERIZATION, FORMATE DEHYDROGENASE, GLYCEROL PRODUCTION, ALCOHOL OXIDASE, SHUTTLE VECTORS, YEAST, GLUCOSE, METABOLISM

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