Assessing Lanthanide-Dependent Methanol Dehydrogenase Activity: The Assay Matters

Manh Tri Phi, Helena Singer, Felix Zäh, Christoph Haisch, Sabine Schneider, Huub J.M. Op den Camp, Lena J. Daumann

Research output: Contribution to journalArticlepeer-review


Artificial dye-coupled assays have been widely adopted as a rapid and convenient method to assess the activity of methanol dehydrogenases (MDH). Lanthanide(Ln)-dependent XoxF-MDHs are able to incorporate different lanthanides (Lns) in their active site. Dye-coupled assays showed that the earlier Lns exhibit a higher enzyme activity than the late Lns. Despite widespread use, there are limitations: oftentimes a pH of 9 and activators are required for the assay. Moreover, Ln-MDH variants are not obtained by isolation from the cells grown with the respective Ln, but by incubation of an apo-MDH with the Ln. Herein, we report the cultivation of Ln-dependent methanotroph Methylacidiphilum fumariolicum SolV with nine different Lns, the isolation of the respective MDHs and the assessment of the enzyme activity using the dye-coupled assay. We compare these results with a protein-coupled assay using its physiological electron acceptor cytochrome cGJ (cyt cGJ). Depending on the assay, two distinct trends are observed among the Ln series. The specific enzyme activity of La-, Ce- and Pr-MDH, as measured by the protein-coupled assay, exceeds that measured by the dye-coupled assay. This suggests that early Lns also have a positive effect on the interaction between XoxF-MDH and its cyt cGJ thereby increasing functional efficiency.

Original languageEnglish
Article numbere202300811
Issue number5
StatePublished - 1 Mar 2024


  • lanthanide-dependent bacteria
  • lanthanides
  • metalloenzymes
  • methanol dehydrogenase
  • methylotrophy


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