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Aromatic Halogenation by Using Bifunctional Flavin Reductase–Halogenase Fusion Enzymes
Author(s) -
Andorfer Mary C.,
Belsare Ketaki D.,
Girlich Anna M.,
Lewis Jared C.
Publication year - 2017
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201700391
Subject(s) - flavin group , cofactor , substrate (aquarium) , chemistry , bifunctional , active site , biochemistry , reductase , halogenation , combinatorial chemistry , stereochemistry , directed evolution , biocatalysis , protein engineering , enzyme , escherichia coli , heterologous expression , biology , organic chemistry , gene , catalysis , recombinant dna , reaction mechanism , ecology , mutant
The remarkable site selectivity and broad substrate scope of flavin‐dependent halogenases (FDHs) has led to much interest in their potential as biocatalysts. Multiple engineering efforts have demonstrated that FDHs can be tuned for non‐native substrate scope and site selectivity. FDHs have also proven useful as in vivo biocatalysts and have been successfully incorporated into biosynthetic pathways to build new chlorinated aromatic compounds in several heterologous organisms. In both cases, reduced flavin cofactor, usually supplied by a separate flavin reductase (FR), is required. Herein, we report functional synthetic, fused FDH‐FR proteins containing various FDHs and FRs joined by different linkers. We show that FDH‐FR fusion proteins can increase product titers compared to the individual components for in vivo biocatalysis in Escherichia coli .