Abstract
Objective
To improve the operational stability and reusability of an epoxide hydrolase (EH) for the biosynthesis of optically active epoxides.
Results
A covalently immobilization strategy was employed to improve the stability of Agrobacterium radiobacter EH by using ethylenediamine (EDA)-functionalised epoxy resin LX-1000EP as carrier. Under the optimal conditions, the activity recovery of immobilized enzyme was 72 % and the specific activity was 634 U/g. Immobilized EH exhibited significantly enhanced thermal stability with a half-life of more than 6.8-fold at 50 °C than that of the free enzyme. A gram-scale kinetic resolution of (R,S)-epichlorohydrin using immobilized preparation as biocatalyst was performed and (R)-epichlorohydrin was obtained with 35 % yield and 99 % enantiomeric excess. The immobilized EH showed good operational stability and even after six reactions, it retained >85 % of the initial activity.
Conclusion
The operational stability and recyclability of immobilized EH on an EDA-functionalized epoxy supports demonstrated its potential for producing (R)-epichlorohydrin.
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Acknowledgments
This work was financial supported by National Natural Science Foundation of China (No. 21406205).
Supporting information
Supplementary Fig. 1—Effect of immobilization temperature (a), pH (b), enzyme concentration (c), and immobilization time (d) on the activity of the immobilized enzyme.
Supplementary Fig. 2—1H-NMR spectra of the product [(R)-ECH].
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Zou, SP., Wang, ZC., Qin, C. et al. Covalent immobilization of Agrobacterium radiobacter epoxide hydrolase on ethylenediamine functionalised epoxy supports for biocatalytical synthesis of (R)-epichlorohydrin. Biotechnol Lett 38, 1579–1585 (2016). https://doi.org/10.1007/s10529-016-2135-y
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DOI: https://doi.org/10.1007/s10529-016-2135-y