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Structure-Based Redesign of a Self-Sufficient Flavin-Containing Monooxygenase towards Indigo Production
- Source :
- International Journal of Molecular Sciences, Volume 20, Issue 24, International Journal of Molecular Sciences, 20(24):6148. MDPI AG
- Publication Year :
- 2019
- Publisher :
- Multidisciplinary Digital Publishing Institute, 2019.
-
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.
- Subjects :
- EXPRESSION
MECHANISM
0301 basic medicine
Indoles
Flavin-containing monooxygenase
Flavin group
Indigo Carmine
OXIDATION
01 natural sciences
Article
Catalysis
Indigo
Mixed Function Oxygenases
Inorganic Chemistry
03 medical and health sciences
Escherichia coli
Enzyme kinetics
indigo
Physical and Theoretical Chemistry
monooxygenase
FMO
Molecular Biology
Spectroscopy
Thermostability
Indole test
010405 organic chemistry
Chemistry
Organic Chemistry
General Medicine
Monooxygenase
flavin
Combinatorial chemistry
0104 chemical sciences
Computer Science Applications
MISO library
030104 developmental biology
Structure based
Oxidation-Reduction
Subjects
Details
- Language :
- English
- ISSN :
- 14220067
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....5da5fde5624f9fa23a36d937967343aa
- Full Text :
- https://doi.org/10.3390/ijms20246148