Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity
The discovery and generation of biocatalysts with extended catalytic versatilities are of immense relevance in both chemistry and biotechnology. An enhanced atomistic understanding of enzyme promiscuity, a mechanism through which living systems acquire novel catalytic functions and specificities by...
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doaj-09bf380e3f374d6bb36bc7e6c1f0d2952020-11-24T22:34:40ZengMDPI AGCatalysts2073-43442016-06-01669010.3390/catal6060090catal6060090Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase ActivityCharlotte Kürten0Bengt Carlberg1Per-Olof Syrén2Science for Life Laboratory, School of Biotechnology, Division of Proteomics and Nanobiotechnology, KTH Royal Institute of Technology, 171 21 Stockholm, SwedenScience for Life Laboratory, School of Biotechnology, Division of Proteomics and Nanobiotechnology, KTH Royal Institute of Technology, 171 21 Stockholm, SwedenScience for Life Laboratory, School of Biotechnology, Division of Proteomics and Nanobiotechnology, KTH Royal Institute of Technology, 171 21 Stockholm, SwedenThe discovery and generation of biocatalysts with extended catalytic versatilities are of immense relevance in both chemistry and biotechnology. An enhanced atomistic understanding of enzyme promiscuity, a mechanism through which living systems acquire novel catalytic functions and specificities by evolution, would thus be of central interest. Using esterase-catalyzed amide bond hydrolysis as a model system, we pursued a simplistic in silico discovery program aiming for the identification of enzymes with an internal backbone hydrogen bond acceptor that could act as a reaction specificity shifter in hydrolytic enzymes. Focusing on stabilization of the rate limiting transition state of nitrogen inversion, our mechanism-guided approach predicted that the acyl hydrolase patatin of the α/β phospholipase fold would display reaction promiscuity. Experimental analysis confirmed previously unknown high amidase over esterase activity displayed by the first described esterase machinery with a protein backbone hydrogen bond acceptor to the reacting NH-group of amides. The present work highlights the importance of a fundamental understanding of enzymatic reactions and its potential for predicting enzyme scaffolds displaying alternative chemistries amenable to further evolution by enzyme engineering.http://www.mdpi.com/2073-4344/6/6/90enzyme promiscuityenzyme catalysisbiocatalysisreaction mechanismsmolecular modelingamidaseesterase |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Charlotte Kürten Bengt Carlberg Per-Olof Syrén |
spellingShingle |
Charlotte Kürten Bengt Carlberg Per-Olof Syrén Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity Catalysts enzyme promiscuity enzyme catalysis biocatalysis reaction mechanisms molecular modeling amidase esterase |
author_facet |
Charlotte Kürten Bengt Carlberg Per-Olof Syrén |
author_sort |
Charlotte Kürten |
title |
Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity |
title_short |
Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity |
title_full |
Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity |
title_fullStr |
Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity |
title_full_unstemmed |
Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity |
title_sort |
mechanism-guided discovery of an esterase scaffold with promiscuous amidase activity |
publisher |
MDPI AG |
series |
Catalysts |
issn |
2073-4344 |
publishDate |
2016-06-01 |
description |
The discovery and generation of biocatalysts with extended catalytic versatilities are of immense relevance in both chemistry and biotechnology. An enhanced atomistic understanding of enzyme promiscuity, a mechanism through which living systems acquire novel catalytic functions and specificities by evolution, would thus be of central interest. Using esterase-catalyzed amide bond hydrolysis as a model system, we pursued a simplistic in silico discovery program aiming for the identification of enzymes with an internal backbone hydrogen bond acceptor that could act as a reaction specificity shifter in hydrolytic enzymes. Focusing on stabilization of the rate limiting transition state of nitrogen inversion, our mechanism-guided approach predicted that the acyl hydrolase patatin of the α/β phospholipase fold would display reaction promiscuity. Experimental analysis confirmed previously unknown high amidase over esterase activity displayed by the first described esterase machinery with a protein backbone hydrogen bond acceptor to the reacting NH-group of amides. The present work highlights the importance of a fundamental understanding of enzymatic reactions and its potential for predicting enzyme scaffolds displaying alternative chemistries amenable to further evolution by enzyme engineering. |
topic |
enzyme promiscuity enzyme catalysis biocatalysis reaction mechanisms molecular modeling amidase esterase |
url |
http://www.mdpi.com/2073-4344/6/6/90 |
work_keys_str_mv |
AT charlottekurten mechanismguideddiscoveryofanesterasescaffoldwithpromiscuousamidaseactivity AT bengtcarlberg mechanismguideddiscoveryofanesterasescaffoldwithpromiscuousamidaseactivity AT perolofsyren mechanismguideddiscoveryofanesterasescaffoldwithpromiscuousamidaseactivity |
_version_ |
1725726150238404608 |