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Cited by 21Enzyme Catalysis and ImmobilizationAsymmetric Synthesis and CatalysisEnzyme Production and Characterization

The DOI matches its Crossref record; 35 of the 35 references it cites were found in a registry or an index.

G‐type Halohydrin Dehalogenases Catalyze Ring Opening Reactions of Cyclic Epoxides with Diverse Anionic Nucleophiles**

Open access
Solarczek, Jennifer, Kaspar, Felix, Bauer, Pia, Schallmey, Anett·2022·Chemistry – A European Journal
DOI 10.1002/chem.202202343PMC PMC10099379

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Abstract

Halohydrin dehalogenases are promiscuous biocatalysts, which enable asymmetric ring opening reactions of epoxides with various anionic nucleophiles. However, despite the increasing interest in such asymmetric transformations, the substrate scope of G‐type halohydrin dehalogenases toward cyclic epoxides has remained largely unexplored, even though this subfamily is the only one known to display activity with these sterically demanding substrates. Herein, we report on the exploration of the substrate scope of the two G‐type halohydrin dehalogenases HheG and HheG2 and a newly identified, more thermostable member of the family, HheG3, with a variety of sterically demanding cyclic epoxides and anionic nucleophiles. This work shows that, in addition to azide and cyanide, these enzymes facilitate ring‐opening reactions with cyanate, thiocyanate, formate, and nitrite, significantly expanding the known repertoire of accessible transformations.

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Registry

Crossref · 10.1002/chem.202202343

Cites

35 of 35 references verified0 citation errors · 0 not checkable

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Figures

Scheme 1 Halohydrin dehalogenase‐catalyzed epoxide ring opening with a range of C‐, N‐, O‐, S‐ and halide (X=Cl, Br) nucleophiles.
Scheme 1 Halohydrin dehalogenase‐catalyzed epoxide ring opening with a range of C‐, N‐, O‐, S‐ and halide (X=Cl, Br) nucleophiles.CC BY · PubMed Central
Figure 1 Characterization of G‐type HHDHs. A) Superposition of the five best predicted models [29] for HheG, HheG2 and HheG3 and the experimental crystal structure of HheG (PDB ID 5o30 [22] ) indicates high structural similarity, expect for the flexible 15 amino acid loop region unique to the G‐type HHDHs. The catalytic triad is shown as sticks and reflects the residue positions in the experimental structure of HheG. B) G‐type HHDHs largely lack dehalogenation activity, as determined by a halide release assay, but are highly active in the nucleophilic ring opening of cyclohexene oxide, setting them apart from other known HHDHs. C) Arrhenius plots obtained by differential scanning fluorimetry reveal that HheG3 is much more stable and cosolvent‐resistant than its relatives, which translates to a longer apparent half‐life under typical operational conditions. D) The catalytic promiscuity with azide as the nucleophile enables the three G‐type HHDHs to convert a series of other cyclic epoxides. Please see the Supporting Information for experimental details.
Figure 1 Characterization of G‐type HHDHs. A) Superposition of the five best predicted models [29] for HheG, HheG2 and HheG3 and the experimental crystal structure of HheG (PDB ID 5o30 [22] ) indicates high structural similarity, expect for the flexible 15 amino acid loop region unique to the G‐type HHDHs. The catalytic triad is shown as sticks and reflects the residue positions in the experimental structure of HheG. B) G‐type HHDHs largely lack dehalogenation activity, as determined by a halide release assay, but are highly active in the nucleophilic ring opening of cyclohexene oxide, setting them apart from other known HHDHs. C) Arrhenius plots obtained by differential scanning fluorimetry reveal that HheG3 is much more stable and cosolvent‐resistant than its relatives, which translates to a longer apparent half‐life under typical operational conditions. D) The catalytic promiscuity with azide as the nucleophile enables the three G‐type HHDHs to convert a series of other cyclic epoxides. Please see the Supporting Information for experimental details.CC BY · PubMed Central
Figure 2 Substrate scope of the G‐type HHDHs. Bold bonds indicate relative configurations and wedges represent absolute configurations. Typical reaction conditions were 20 mM epoxide, 40 mM nucleophile, and 200 μg HHDH in 1 mL of 50 mM Tris⋅SO4 buffer with 2 % iPrOH at pH 7 and 22 °C. TTNs were determined by analysis of crude reaction mixtures by achiral GC and refer to the sum of all formed product isomers. The ratios refer to the ratio of formed regioisomers (e. g. 4 a vs. 4 a′), except for reactions with thiocyanate, where given ratios refer to the ratio of isomers arising from S : N‐nucleophilic attack. These values are normalized to the isomer shown. Enantiomeric excesses were determined by analysis of the crude reaction mixtures by chiral GC and also refer to the isomer shown. The ee values in parentheses refer to the corresponding other (regio)isomer. Please see the Supporting Information for the full experimental details and analytical procedures.
Figure 2 Substrate scope of the G‐type HHDHs. Bold bonds indicate relative configurations and wedges represent absolute configurations. Typical reaction conditions were 20 mM epoxide, 40 mM nucleophile, and 200 μg HHDH in 1 mL of 50 mM Tris⋅SO4 buffer with 2 % iPrOH at pH 7 and 22 °C. TTNs were determined by analysis of crude reaction mixtures by achiral GC and refer to the sum of all formed product isomers. The ratios refer to the ratio of formed regioisomers (e. g. 4 a vs. 4 a′), except for reactions with thiocyanate, where given ratios refer to the ratio of isomers arising from S : N‐nucleophilic attack. These values are normalized to the isomer shown. Enantiomeric excesses were determined by analysis of the crude reaction mixtures by chiral GC and also refer to the isomer shown. The ee values in parentheses refer to the corresponding other (regio)isomer. Please see the Supporting Information for the full experimental details and analytical procedures.CC BY · PubMed Central
Figure 3 Illustrative analytical data for selected HHDH‐catalyzed ring‐opening reactions. *internal standard.
Figure 3 Illustrative analytical data for selected HHDH‐catalyzed ring‐opening reactions. *internal standard.CC BY · PubMed Central

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Citations

Cites

Research works printed but not linked: 3

  1. 10.1038/s41592-022-01488-1 · DOI · Paper page
  2. Arabnejad H., 2017, Protein Eng. Des. Sel., 30, 175
  3. Wang H., 2022, Angew. Chem., 202205790

Metadata

Authors
Solarczek, J., Kaspar, F., Bauer, P., & Schallmey, A.
Affiliations
  • Technische Universität Braunschweig (DE)
  • Technische Universität Berlin (DE)
  • Amedes Genetics (Germany) (DE)
Corresponding author
Schallmey, Anett
ORCID
Venue
Chemistry – A European Journal
Publisher
Wiley
Published
2022-11-14
Volume
28
Issue
72
Pages
e202202343-e202202343
ISSN
  • 0947-6539 (ISSN-L)
  • 1521-3765
DOI
10.1002/chem.202202343
PMC
PMC10099379
OpenAlex
W4303986034
Cited by
21
References
35
Citations by year
2026: 5 · 2025: 5 · 2024: 7 · 2023: 2 · 2022: 2
FWCI
1.31
Type
journal article
Language
en
Open access
Licence
Locations
  • Chemistry - A European Journal (publishedVersion, cc-by)
  • ChemRxiv (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DepositOnce (submittedVersion, cc-by)
  • Technische Universität Berlin – Universitätsbibliothek (cc-by)
Funders
Deutsche Forschungsgemeinschaft (SCHA 1745/2-2, 492196858)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
  • Physical Sciences › Chemistry › Organic Chemistry › Asymmetric Synthesis and Catalysis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
Keywords
epoxide ring opening, anionic nucleophiles, asymmetric biocatalysis, cyanate, thiocyanate, azide, cyanide, nitrite, formate, thermostable enzymes, substrate scope
MeSH terms
Catalysis, Epoxy Compounds, Hydrolases, Nitrites

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