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Nanofructosome encapsulated CalB enzyme immobilized silica-coated magnetic nanoparticles for rapid enzymatic hydrolysis and acylation

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Abstract

This study reports the preparation of Candida Antarctic lipase B (CalB) enzyme and nanofructosome encapsulated CalB enzyme (CalB@NF) immobilized silica coated magnetic nanoparticles (Si-MNPs) and demonstration of the enzymatic hydrolysis and acylation against p-nitrophenyl butyrate (p-NPB) and benzoic anhydride. Si-MNPs was prepared 60 nm of particle size with spherical shape, and the immobilization of CalB and CalB@NF was coupled by chlorosilane linker on the surface of Si-MNPs. The quantitative determination of CalB in Si-MNPs@CalB and Si-MNPs@CalB@NF was followed by Bradford assay. Various enzymatic kinetic parameters such as Km, Vmax, and Kcat were calculated using the Lineweaver-Burk equation and Michaelis-Menten kinetics. The hydrolysis of p-NPB with Si-MNPs@CalB@NF and Si-MNPS@CalB confirmed their utilization as effective catalysts. Furthermore, all four samples of native CalB, CalB@NF, Si-MNPs@CalB, and Si-MNPs@CalB@NF were successfully acylated to benzyl benzoate after column separation, and showed more than 99% conversion efficiency. The stability for pH and temperature of Si-MNPs@CalB and Si-MNPs@CalB@NF was optimized at 8 and 45 ℃, respectively.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was carried out with the support of “Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ01493802)” Rural Development Administration, Republic of Korea.

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  1. Korea Institute of Ceramic Engineering and Technology, Chungbuk, 28160, Korea

    Yu Jeong Kim, Woo Young Jang & Jeong Ho Chang

  2. Department of Chemical Engineering, Yonsei University, Seoul, 03722, Korea

    Yu Jeong Kim

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Kim, Y., Jang, W. & Chang, J.H. Nanofructosome encapsulated CalB enzyme immobilized silica-coated magnetic nanoparticles for rapid enzymatic hydrolysis and acylation. J. Korean Ceram. Soc. 61, 126–136 (2024). https://doi.org/10.1007/s43207-023-00355-9

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