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Engineering Orthogonal Methyltransferases to Create Alternative Bioalkylation Pathways
Author(s) -
Herbert Abigail J.,
Shepherd Sarah A.,
Cronin Victoria A.,
Bennett Matthew R.,
Sung Rehana,
Micklefield Jason
Publication year - 2020
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.202004963
Subject(s) - methyltransferase , tetrahydroisoquinoline , chemistry , enzyme , methylation , stereochemistry , catechol , mutagenesis , cofactor , stereoselectivity , biocatalysis , ethionine , combinatorial chemistry , biochemistry , methionine , catalysis , amino acid , gene , mutation , reaction mechanism
S ‐adenosyl‐ l ‐methionine (SAM)‐dependent methyltransferases (MTs) catalyse the methylation of a vast array of small metabolites and biomacromolecules. Recently, rare carboxymethylation pathways have been discovered, including carboxymethyltransferase enzymes that utilise a carboxy‐SAM (cxSAM) cofactor generated from SAM by a cxSAM synthase (CmoA). We show how MT enzymes can utilise cxSAM to catalyse carboxymethylation of tetrahydroisoquinoline (THIQ) and catechol substrates. Site‐directed mutagenesis was used to create orthogonal MTs possessing improved catalytic activity and selectivity for cxSAM, with subsequent coupling to CmoA resulting in more efficient and selective carboxymethylation. An enzymatic approach was also developed to generate a previously undescribed co‐factor, carboxy‐ S ‐adenosyl‐ l ‐ethionine (cxSAE), thereby enabling the stereoselective transfer of a chiral 1‐carboxyethyl group to the substrate.