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Evolved Aliphatic Halogenases Enable Regiocomplementary C−H Functionalization of a Pharmaceutically Relevant Compound
Author(s) -
Hayashi Takahiro,
Ligibel Mathieu,
Sager Emine,
Voss Moritz,
Hunziker Jürg,
Schroer Kirsten,
Snajdrova Radka,
Buller Rebecca
Publication year - 2019
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201907245
Subject(s) - halogenation , regioselectivity , chemistry , directed evolution , substrate (aquarium) , stereochemistry , enzyme , protein engineering , active site , combinatorial chemistry , heme , halogen , biochemistry , organic chemistry , biology , catalysis , ecology , alkyl , mutant , gene
Non‐heme iron halogenases are synthetically valuable biocatalysts that are capable of halogenating unactivated sp 3 ‐hybridized carbon centers with high stereo‐ and regioselectivity. The reported substrate scope of these enzymes, however, is limited primarily to the natural substrates and their analogues. We engineered the halogenase WelO5* for chlorination of a martinelline‐derived fragment. Using structure‐guided evolution, a halogenase variant with a more than 290‐fold higher total turnover number and a 400‐fold higher apparent k cat compared to the wildtype enzyme was generated. Moreover, we identified key positions in the active site that allow direction of the halogen to different positions in the target substrate. This is the first example of enzyme engineering to expand the substrate scope of a non‐heme iron halogenase beyond the native indole‐alkaloid‐type substrates. The highly evolvable nature of WelO5* underscores the usefulness of this enzyme family for late‐stage halogenation.