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Discovery and Design of Family VIII Carboxylesterases as Highly Efficient Acyltransferases
Author(s) -
Müller Henrik,
Godehard Simon P.,
Palm Gottfried J.,
Berndt Leona,
Badenhorst Christoffel P. S.,
Becker AnnKristin,
Lammers Michael,
Bornscheuer Uwe T.
Publication year - 2021
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.202014169
Subject(s) - acyltransferases , chemistry , carboxylesterase , acyltransferase , hydrolase , esterase , hydrolysis , biochemistry , stereochemistry , active site , enzyme , biosynthesis
Promiscuous acyltransferase activity is the ability of certain hydrolases to preferentially catalyze acyl transfer over hydrolysis, even in bulk water. However, poor enantioselectivity, low transfer efficiency, significant product hydrolysis, and limited substrate scope represent considerable drawbacks for their application. By activity‐based screening of several hydrolases, we identified the family VIII carboxylesterase, EstCE1, as an unprecedentedly efficient acyltransferase. EstCE1 catalyzes the irreversible amidation and carbamoylation of amines in water, which enabled the synthesis of the drug moclobemide from methyl 4‐chlorobenzoate and 4‐(2‐aminoethyl)morpholine (ca. 20 % conversion). We solved the crystal structure of EstCE1 and detailed structure–function analysis revealed a three‐amino acid motif important for promiscuous acyltransferase activity. Introducing this motif into an esterase without acetyltransferase activity transformed a “hydrolase” into an “acyltransferase”.