4UFO

Laboratory evolved variant R-C1B1D33E6 of potato epoxide hydrolase StEH1

Summary for 4UFO

• Entry DOI: 10.2210/pdb4ufo/pdb
• Related: 4UFN 4UFP
• Descriptor: EPOXIDE HYDROLASE (2 entities in total)
• Functional Keywords: hydrolase, epoxide hydrolysis, alpha/beta-hydrolase, directed evolution, asymmetric syntheses
• Biological source: SOLANUM TUBEROSUM (POTATO)
• Total number of polymer chains: 2
• Total formula weight: 74148.90

Authors

Carlsson, A.J.,Bauer, P.,Nilsson, M.,Dobritzsch, D.,Kamerlin, S.C.L.,Widersten, M. (deposition date: 2015-03-17, release date: 2016-04-13, Last modification date: 2023-12-20)

Primary citation

Janfalk Carlsson, A.,Bauer, P.,Dobritzsch, D.,Nilsson, M.,Kamerlin, S.C.,Widersten, M.
Laboratory Evolved Enzymes Provide Snapshots of the Development of Enantioconvergence in Enzyme-Catalyzed Epoxide Hydrolysis.
Chembiochem, 17:1693-, 2016

PubMed Abstract: Engineered enzyme variants of potato epoxide hydrolase (StEH1) display varying degrees of enrichment of (2R)-3-phenylpropane-1,2-diol from racemic benzyloxirane. Curiously, the observed increase in the enantiomeric excess of the (R)-diol is not only a consequence of changes in enantioselectivity for the preferred epoxide enantiomer, but also to changes in the regioselectivity of the epoxide ring opening of (S)-benzyloxirane. In order to probe the structural origin of these differences in substrate selectivity and catalytic regiopreference, we solved the crystal structures for the evolved StEH1 variants. We used these structures as a starting point for molecular docking studies of the epoxide enantiomers into the respective active sites. Interestingly, despite the simplicity of our docking analysis, the apparent preferred binding modes appear to rationalize the experimentally determined regioselectivities. The analysis also identifies an active site residue (F33) as a potentially important interaction partner, a role that could explain the high conservation of this residue during evolution. Overall, our experimental, structural, and computational studies provide snapshots into the evolution of enantioconvergence in StEH1-catalyzed epoxide hydrolysis.

PubMed: 27383542
DOI: 10.1002/CBIC.201600330

Experimental method

X-RAY DIFFRACTION (2.02 Å)