Asymmetric sulfoxidation by engineering the heme pocket of a dye-decolorizing peroxidase

Cita com:
hdl:2117/87761
Document typeArticle
Defense date2016-05-23
PublisherRoyal Society of Chemistry
Rights accessOpen Access
Except where otherwise noted, content on this work
is licensed under a Creative Commons license
:
Attribution-NonCommercial-NoDerivs 4.0 Spain
ProjectINDOX - Optimized oxidoreductases for medium and large scale industrial biotransformations (EC-FP7-613549)
NUEVAS ENZIMAS OXIDATIVAS PARA UNA INDUSTRIA SOSTENIBLE (MINECO-BIO2014-56388-R)
DESENMASCARANDO PATOGENICIDAD Y VIRULENCIA EN ACINETOBACTER BAUMANNII: UNA APROXIMACION ESTRUCTURAL (MINECO-BFU2014-55448-P)
DISENYO COMPUTACIONAL RACIONAL DE OXIDOREDUCTASAS PARA APLICACIONES INDUSTRIALES Y TECNOLOGICAS (MINECO-CTQ2013-48287-R)
NUEVAS ENZIMAS OXIDATIVAS PARA UNA INDUSTRIA SOSTENIBLE (MINECO-BIO2014-56388-R)
DESENMASCARANDO PATOGENICIDAD Y VIRULENCIA EN ACINETOBACTER BAUMANNII: UNA APROXIMACION ESTRUCTURAL (MINECO-BFU2014-55448-P)
DISENYO COMPUTACIONAL RACIONAL DE OXIDOREDUCTASAS PARA APLICACIONES INDUSTRIALES Y TECNOLOGICAS (MINECO-CTQ2013-48287-R)
Abstract
The so-called dye-decolorizing peroxidases (DyPs) constitute a new family of proteins exhibiting remarkable stability. With the aim of providing them new catalytic activities of biotechnological interest, the heme pocket of one of the few DyPs fully characterized to date (from the fungus Auricularia auricula-judae) was redesigned based on the crystal structure available, and its potential for asymmetric sulfoxidation was evaluated. Chiral sulfoxides are important targets in organic synthesis and enzyme catalysis, due to a variety of applications. Interestingly, one of the DyP variants, F359G, is highly stereoselective in sulfoxidizing methylphenyl sulfide and methyl-p-tolyl sulfide (95–99% conversion, with up to 99% excess of the S enantiomer in short reaction times), while the parent DyP has no sulfoxidation activity, and the L357G variant produces both R and S enantiomers. The two variants were crystallized, and their crystal structures were used in molecular simulations to provide a rational explanation for the new catalytic activities. Protein energy landscape exploration (PELE) showed more favorable protein–substrate catalytic complexes for the above variants, with a considerable number of structures near the oxygen atom of the activated heme, which is incorporated into the substrates as shown in 18O-labeling experiments, and improved affinity with respect to the parent enzyme, explaining their sulfoxidation activity. Additional quantum mechanics/molecular mechanics (QM/MM) calculations were performed to elucidate the high stereoselectivity observed for the F359G variant, which correlated with higher reactivity on the substrate molecules adopting pro-S poses at the active site. Similar computational analyses can help introduce/improve (stereoselective) sulfoxidation activity in related hemeproteins.
CitationLinde, Dolores [et al.]. Asymmetric sulfoxidation by engineering the heme pocket of a dye-decolorizing peroxidase. "Catalysis Science & Technology", 23 Maig 2016.
ISSN2044-4753
Collections
Files | Description | Size | Format | View |
---|---|---|---|---|
Asymmetric sulfoxidation by engineering the.pdf | 3,439Mb | View/Open |