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Covalent Immobilization of Catalase onto Regenerated Silk Fibroins via Tyrosinase-Catalyzed Cross-Linking

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Abstract

Regenerated silk fibroins could be used as medical scaffolds and carrier materials for enzyme immobilization. In the present work, tyrosinase enzyme was used for enzymatic oxidation of silk fibroins, followed by immobilization of catalase onto the fibroin surfaces through physical adsorption and covalent cross-linking as well. Spectrophotometry, SDS-PAGE, and Fourier transform infrared spectroscopy (FTIR) were used to examine the efficiency of enzymatic oxidation and catalase immobilization, respectively. The results indicate that tyrosine residues in silk fibroins could be oxidized and converted to the active o-quinones. Incubating silk fibroins with catalase and tyrosinase led to a noticeable change of molecular weight distribution, indicating the occurrence of the cross-links between silk fibroins and catalase molecules. Two different pathways were proposed for the catalase immobilizations, and the method based on grafting of catalase onto the freeze-dried fibroin membrane is more acceptable. The residual enzyme activity for the immobilized catalase exhibited higher than that of the control after repeated washing cycles. Meanwhile, the thermal stability and alkali resistance were also slightly improved as compared to free catalase. The mechanisms of enzymatic immobilization are also concerned.

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Acknowledgments

This work was financially supported by the National Natural Science Funds (51373071), Program for New Century Excellent Talents in University (NCET-12-0883), and the Fundamental Research Funds for the Central Universities (JUSRP51312B).

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Authors and Affiliations

  1. Key Laboratory of Science and Technology of Eco-Textile, Ministry of Education, Jiangnan University, Wuxi, 214122, People’s Republic of China

    Ping Wang, Chenglong Qi, Yuanyuan Yu, Jiugang Yuan, Li Cui, Gengtie Tang, Qiang Wang & Xuerong Fan

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  1. Ping Wang
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  2. Chenglong Qi
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  3. Yuanyuan Yu
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  4. Jiugang Yuan
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  8. Xuerong Fan
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Correspondence to Ping Wang.

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Wang, P., Qi, C., Yu, Y. et al. Covalent Immobilization of Catalase onto Regenerated Silk Fibroins via Tyrosinase-Catalyzed Cross-Linking. Appl Biochem Biotechnol 177, 472–485 (2015). https://doi.org/10.1007/s12010-015-1756-2

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  • DOI: https://doi.org/10.1007/s12010-015-1756-2

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