Structure-Based Redesign of a Self-Sufficient Flavin-Containing Monooxygenase towards Indigo Production

Nikola Lončar*, Hugo L. van Beek, Marco W. Fraaije

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

4 Citations (Scopus)
75 Downloads (Pure)

Abstract

Indigo is currently produced by a century-old petrochemical-based process, therefore it is highly attractive to develop a more environmentally benign and efficient biotechnological process to produce this timeless dye. Flavin-containing monooxygenases (FMOs) are able to oxidize a wide variety of substrates. In this paper we show that the bacterial mFMO can be adapted to improve its ability to convert indole into indigo. The improvement was achieved by a combination of computational and structure-inspired enzyme redesign. We showed that the thermostability and the kcat for indole could be improved 1.5-fold by screening a relatively small number of enzyme mutants. This project not only resulted in an improved biocatalyst but also provided an improved understanding of the structural elements that determine the activity of mFMO and provides hints for further improvement of the monooxygenase as biocatalyst.

Original languageEnglish
Article number6148
Number of pages9
JournalInternational Journal of Molecular Sciences
Volume20
Issue number24
DOIs
Publication statusPublished - 5-Dec-2019

Keywords

  • indigo
  • MISO library
  • flavin
  • monooxygenase
  • FMO
  • EXPRESSION
  • OXIDATION
  • MECHANISM

Cite this