Self-assembly of metal-cholesterol oxidase hybrid nanostructures and application in bioconversion of steroids derivatives

Abstract

A cholesterol oxidase (COD) was hybridized with Ca²⁺, Zn²⁺, Al³⁺, Fe²⁺ and Mn²⁺. After precipitation with PO₄^3– at 4 °C for 72 h, the resulting pellets were freeze-dried. In scanning electron microscopy assays, the metal-COD complexes revealed flower-like or granular structures after hybridization. Fourier transform infrared spectroscopy assay revealed the characteristic peaks of both the enzyme and metal materials. X-ray diffraction analysis indicated that COD was encapsulated in CaHPO₄·2H₂O-, Zn₃(PO₄)₂·4H₂O-, AlPO₄-, FeP₄- and Mn₃(PO₄)₂·3H₂O-based nanostructures, respectively. Differential scanning calorimetry assay indicated significant increases in thermo-denaturation temperatures from 60.5 °C to 167.02 °C, 167.02 °C, 137.70 °C, 172.85 °C and 160.99 °C, respectively. Using steroid derivatives as substrates, this enzyme could convert cholesterol, pregnenolone, dehydroepiandrosterone, ergosterol, b-sitosterol and stigmasterol to related single products. Hybridization in metal-based nanostructures could significantly enhance the initial conversion ratio and reaction stability of the enzyme. In addition, substrate selectivity could be affected by various metal materials. Briefly, using Ca²⁺, Zn²⁺, Al³⁺, Fe²⁺ and Mn²⁺ as hybrid raw materials could help to encapsulate COD in related metal-enzyme nanostructures, and could help to promote the stability and tolerant properties of the enzyme, while also enhancing its catalytic characteristics.

Key words： cholesterol oxidase; metal-enzyme hybridization; nanostructures; sterol derivatives; bioconversion

Acknowledgement

This work was supported by the National Key Research and Development Program of China (Grant No. 2018YFA0900304-300); the Natural Science Foundation of Jiangsu Province (Grant Nos. BK20160053 and BE2018055); the Priority Academic Program Development of Jiangsu Higher Education Institutions, 111 Project (Grant No. 111-2-06).

Electronic Supplementary Material

Supplementary material is available in the online version of this article at https://doi.org/10.1007/s11705-020-1989-7 and is accessible for authorized users.

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