Sökning: onr:"swepub:oai:DiVA.org:kth-189936" >
Mechanism-Guided Di...
Mechanism-Guided Discovery of an Esterase Scaffold with Promiscuous Amidase Activity
-
- Kürten, Charlotte (författare)
- KTH,Proteomik och nanobioteknologi
-
- Carlberg, Bengt (författare)
- KTH,Proteomik och nanobioteknologi
-
- Syren, Per-Olof (författare)
- KTH,Proteomik och nanobioteknologi
-
(creator_code:org_t)
- 2016-06-18
- 2016
- Engelska.
-
Ingår i: Catalysts. - : MDPI AG. - 2073-4344. ; 6:6
- Relaterad länk:
-
https://www.mdpi.com...
-
visa fler...
-
https://urn.kb.se/re...
-
https://doi.org/10.3...
-
visa färre...
Abstract
Ämnesord
Stäng
- 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 alpha/beta 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.
Nyckelord
- enzyme promiscuity
- enzyme catalysis
- biocatalysis
- reaction mechanisms
- molecular modeling
- amidase
- esterase
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas