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Sequence‐Based Prediction of Promiscuous Acyltransferase Activity in Hydrolases
Author(s) -
Müller Henrik,
Becker AnnKristin,
Palm Gottfried J.,
Berndt Leona,
Badenhorst Christoffel P. S.,
Godehard Simon P.,
Reisky Lukas,
Lammers Michael,
Bornscheuer Uwe T.
Publication year - 2020
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.202003635
Subject(s) - acyltransferases , acyltransferase , substrate specificity , chemistry , biochemistry , sequence (biology) , peptide sequence , computational biology , active site , enzyme , substrate (aquarium) , biology , biosynthesis , ecology , gene
Certain hydrolases preferentially catalyze acyl transfer over hydrolysis in an aqueous environment. However, the molecular and structural reasons for this phenomenon are still unclear. Herein, we provide evidence that acyltransferase activity in esterases highly correlates with the hydrophobicity of the substrate‐binding pocket. A hydrophobicity scoring system developed in this work allows accurate prediction of promiscuous acyltransferase activity solely from the amino acid sequence of the cap domain. This concept was experimentally verified by systematic investigation of several homologous esterases, leading to the discovery of five novel promiscuous acyltransferases. We also developed a simple yet versatile colorimetric assay for rapid characterization of novel acyltransferases. This study demonstrates that promiscuous acyltransferase activity is not as rare as previously thought and provides access to a vast number of novel acyltransferases with diverse substrate specificity and potential applications.