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Covalent immobilization of β-1,4-glucosidase from Agaricus arvensis onto functionalized silicon oxide nanoparticles

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Abstract

An efficient β-1,4-glucosidase (BGL) secreting strain, Agaricus arvensis, was isolated and identified. The relative molecular weight of the purified A. arvensis BGL was 98 kDa, as determined by sodium dodecylsulfate polyacrylamide gel electrophoresis, or 780 kDa by size exclusion chromatography, indicating that the enzyme is an octamer. Using a crude enzyme preparation, A. arvensis BGL was covalently immobilized onto functionalized silicon oxide nanoparticles with an immobilization efficiency of 158%. The apparent V max (k cat) values of free and immobilized BGL under standard assay conditions were 3,028 U mg protein−1 (4,945 s−1) and 3,347 U mg protein−1 (5,466 s−1), respectively. The immobilized BGL showed a higher optimum temperature and improved thermostability as compared to the free enzyme. The half-life at 65 °C showed a 288-fold improvement over the free BGL. After 25 cycles, the immobilized enzyme still retained 95% of the original activity, thus demonstrating its prospects for commercial applications. High specific activity, high immobilization efficiency, improved stability, and reusability of A. arvensis BGL make this enzyme of potential interest in a number of industrial applications.

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Acknowledgment

This study was supported by a grant (code 2008A0080126) from ARPC, Republic of Korea. This work was also supported by a grant (code 20070301034024) from Biogreen 21 Program, Rural Development Administration, Republic of Korea.

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  1. Department of Chemical Engineering, Konkuk University, 1 Hwayang-Dong, Gwangjin-Gu, Seoul, South Korea, 143-701

    Raushan Kumar Singh, Ye-Wang Zhang, Ngoc-Phuong-Thao Nguyen, Marimuthu Jeya & Jung-Kul Lee

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  1. Raushan Kumar Singh
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  2. Ye-Wang Zhang
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  3. Ngoc-Phuong-Thao Nguyen
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  4. Marimuthu Jeya
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  5. Jung-Kul Lee
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Correspondence to Marimuthu Jeya or Jung-Kul Lee.

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Singh, R.K., Zhang, YW., Nguyen, NPT. et al. Covalent immobilization of β-1,4-glucosidase from Agaricus arvensis onto functionalized silicon oxide nanoparticles. Appl Microbiol Biotechnol 89, 337–344 (2011). https://doi.org/10.1007/s00253-010-2768-z

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  • DOI: https://doi.org/10.1007/s00253-010-2768-z

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