In:
ChemBioChem, Wiley, Vol. 24, No. 3 ( 2023-02)
Abstract:
Epoxy‐norbornane (EPO‐NBE) is a crucial building block for the synthesis of various biologically active heterocyclic systems. To develop an efficient protocol for producing EPO‐NBE using norbornene (NBE) as a substrate, cytochrome P450 enzyme from Pseudomonas putida (CYP238A1) was examined and its crystal structure (PDB code: 7X53) was resolved. Molecular mechanism analysis showed a high energy barrier related to iron‐alkoxy radical complex formation. Therefore, a protein engineering strategy was developed and an optimal CYP238A1 NPV variant containing a local hydrophobic “fence” at the active site was obtained, which increased the H 2 O 2 ‐dependent epoxidation activity by 7.5‐fold compared with that of CYP238A1 WT . Among the “fence”, Glu255 participates in an efficient proton transfer system. Whole‐cell transformation using CYP238A1 NPV achieved an EPO‐NBE yield of 77.6 g ⋅ L −1 in a 30‐L reactor with 66.3 % conversion. These results demonstrate the potential of this system for industrial production of EPO‐NBE and provides a new biocatalytic platform for epoxidation chemistry.
Type of Medium:
Online Resource
ISSN:
1439-4227
,
1439-7633
DOI:
10.1002/cbic.202200529
Language:
English
Publisher:
Wiley
Publication Date:
2023
detail.hit.zdb_id:
2020469-3
SSG:
12
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