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Directed Evolution of RebH for Site‐Selective Halogenation of Large Biologically Active Molecules
Author(s) -
Payne James T.,
Poor Catherine B.,
Lewis Jared C.
Publication year - 2015
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201411901
Subject(s) - halogenation , substrate (aquarium) , directed evolution , biocatalysis , chemistry , combinatorial chemistry , active site , stereochemistry , enzyme , catalysis , organic chemistry , biology , reaction mechanism , biochemistry , mutant , ecology , gene
We recently characterized the substrate scope of wild‐type RebH and proceeded to evolve variants of this enzyme with improved stability for biocatalysis. The substrate scopes of both RebH and the stabilized variants, however, are limited primarily to compounds similar in size to tryptophan. A substrate walking approach was used to further evolve RebH variants with expanded substrate scope. Two particularly notable variants were identified: 3‐SS, which provides high conversion of tricyclic tryptoline derivatives; and 4‐V, which accepts a broad range of large indoles and carbazoles. This constitutes the first reported use of directed evolution to enable the functionalization of substrates not accepted by wild‐type RebH and demonstrates the utility of RebH variants for the site‐selective halogenation of biologically active compounds.