Mutagenesis-Independent Stabilization of Class B Flavin Monooxygenases in Operation

Leticia C.P. Goncalves, Daniel Kracher, Sofia Milker, Michael J. Fink*, Florian Rudroff, Roland Ludwig, Andreas S. Bommarius, Marko D. Mihovilovic

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

This paper describes the stabilization of flavin-dependent monooxygenases under reaction conditions, using an engineered formulation of additives (the natural cofactors NADPH and FAD, and superoxide dismutase and catalase as catalytic antioxidants). This way, a 103- to 104-fold increase of the half-life was reached without resource-intensive directed evolution or structure-dependent protein engineering methods. The stabilized enzymes are highly valued for their synthetic potential in biotechnology and medicinal chemistry (enantioselective sulfur, nitrogen and Baeyer–Villiger oxidations; oxidative human metabolism), but widespread application was so far hindered by their notorious fragility. Our technology immediately enables their use, does not require structural knowledge of the biocatalyst, and creates a strong basis for the targeted development of improved variants by mutagenesis. (Figure presented.).

Original languageEnglish
Pages (from-to)2121-2131
Number of pages11
JournalAdvanced Synthesis and Catalysis
Volume359
Issue number12
DOIs
Publication statusPublished - 19 Jun 2017
Externally publishedYes

Keywords

  • biocatalysis
  • cofactors
  • enzyme stabilization
  • oxygenation
  • reactive oxygen species

ASJC Scopus subject areas

  • Catalysis
  • Organic Chemistry

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