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Engineered RebH Halogenase Variants Demonstrating a Specificity Switch from Tryptophan towards Novel Indole Compounds
- Source :
- Chembiochem
- Publication Year :
- 2021
- Publisher :
- Wiley, 2021.
-
Abstract
- Activating industrially important aromatic hydrocarbons by installing halogen atoms is extremely important in organic synthesis and often improves the pharmacological properties of drug molecules. To this end, tryptophan halogenase enzymes are potentially valuable tools for regioselective halogenation of arenes, including various industrially important indole derivatives and similar scaffolds. Although endogenous enzymes show reasonable substrate scope towards indole compounds, their efficacy can often be improved by engineering. Using a structure‐guided semi‐rational mutagenesis approach, we have developed two RebH variants with expanded biocatalytic repertoires that can efficiently halogenate several novel indole substrates and produce important pharmaceutical intermediates. Interestingly, the engineered enzymes are completely inactive towards their natural substrate tryptophan in spite of their high tolerance to various functional groups in the indole ring. Computational modelling and molecular dynamics simulations provide mechanistic insights into the role of gatekeeper residues in the substrate binding site and the dramatic switch in substrate specificity when these are mutated.<br />Two engineered halogenase enzymes were developed by switching substrate specificity of the parental enzyme from tryptophan to indole compounds. They can halogenate several industrially important indole derivatives more efficiently than the parental enzyme; this includes enzymatic chlorination/bromination of novel substrates that were not reported previously. The engineered enzymes could be useful in regioselective synthesis of drug intermediates.
- Subjects :
- Indoles
Halogenation
biocatalysis
Mutagenesis (molecular biology technique)
Molecular Dynamics Simulation
Biochemistry
Substrate Specificity
chemistry.chemical_compound
Bacterial Proteins
directed evolution
Molecular Biology
Indole test
Binding Sites
Full Paper
Chemistry
Organic Chemistry
Tryptophan
Protein engineering
Full Papers
Directed evolution
Combinatorial chemistry
Actinobacteria
halogenase
enzyme engineering
indole
Biocatalysis
Mutagenesis, Site-Directed
Molecular Medicine
Organic synthesis
RebH
Oxidoreductases
Subjects
Details
- ISSN :
- 14397633 and 14394227
- Volume :
- 22
- Database :
- OpenAIRE
- Journal :
- ChemBioChem
- Accession number :
- edsair.doi.dedup.....fda21bf4bf9ba1f3a8bd0a81def06f3f