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Novel immobilization process of a thermophilic catalase: efficient purification by heat treatment and subsequent immobilization at high temperature

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Abstract

The main goal of the present work is to investigate a novel process of purification and immobilization of a thermophilic catalase at high temperatures. The catalase, originated from Bacillus sp., was overexpressed in a recombinant Escherichia coli BL21(DE3)/pET28-CATHis and efficiently purified by heat treatment, achieving a threefold purification. The purified catalase was then immobilized onto an epoxy support at different temperatures (25, 40, and 55 °C). The immobilizate obtained at higher temperatures reached its maximum activity in a shorter time than that obtained at lower temperatures. Furthermore, immobilization at higher temperatures required a lower ionic strength than immobilization at lower temperatures. The characteristics of immobilized enzymes prepared at different temperatures were investigated. The high-temperature immobilizate (55 °C) showed the highest thermal stability, followed by the 40 °C immobilizate. And the high-temperature immobilizate (55 °C) had slightly higher operational stability than the 25 °C immobilizate. All of the immobilized catalase preparations showed higher stability than the free enzyme at alkaline pH 10.0, while the alkali resistance of the 25 °C immobilizate was slightly better than that of the 40 and 55 °C immobilizates.

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Acknowledgments

This work was supported by the National Science Foundation of China (21276023; 21476025), Beijing Natural Science Foundation (2143041), and the Fundamental Research Funds for the Central Universities of China (FRF-BR-11-002A; FRF-SD-12-007A). The authors thank Sunresin New Materials Co., Ltd. for the kind gift of LX-1000EPC4 support. The authors have no conflict of interest to declare.

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Authors and Affiliations

  1. School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, 100083, China

    Juan Xu & Hui Luo

  2. School of Civil and Environmental Engineering, University of Science and Technology Beijing, Beijing, 100083, China

    Juan Xu, Claudia López, Jing Xiao & Yanhong Chang

  3. Key Laboratory of Educational Ministry for High Efficient Mining and Safety in Metal Mine, University of Science and Technology Beijing, Beijing, 100083, China

    Juan Xu, Claudia López, Jing Xiao & Yanhong Chang

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  1. Juan Xu
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  2. Hui Luo
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  5. Yanhong Chang
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Correspondence to Hui Luo or Yanhong Chang.

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Xu, J., Luo, H., López, C. et al. Novel immobilization process of a thermophilic catalase: efficient purification by heat treatment and subsequent immobilization at high temperature. Bioprocess Biosyst Eng 38, 1983–1991 (2015). https://doi.org/10.1007/s00449-015-1439-y

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  • DOI: https://doi.org/10.1007/s00449-015-1439-y

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