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Novel magnetic microspheres of P (GMA-b-HEMA): preparation, lipase immobilization and enzymatic activity in two phases

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Abstract

Magnetic oleic-acid-coated Fe3O4 nanoparticles were first introduced into 1, 1-diphenylethylene (DPE)-controlled radical polymerization system to prepare superparamagnetic microspheres for enzyme immobilization by two steps of polymerization. In the presence of DPE, glycidyl methacrylate, 2-hydroxyethyl methacrylate and methacryloxyethyl trimethyl ammonium chloride with charge were selected as copolymering monomers based on their reactive functional group and excellent biocompatibility which were suitable for immobilization of Candida rugosa lipase (CRL). The resulting magnetic microspheres were characterized by means of scanning electron microscope, Fourier transform infrared spectrum, thermogravimetric analysis and vibrating sample magnetometry. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis SDS-PAGE analysis was also conducted to demonstrate whether CRL is covalently immobilized or only physically adsorbed. The results indicated that the polymerization was successfully carried out, and lipase was immobilized on the magnetic microspheres through ionic adsorption and covalent binding under mild conditions. The immobilized lipase exhibited high activity recovery (69.7%), better resistance to pH and temperature inactivation in aqueous phase, as well as superior reusability in nonaqueous phase. The data showed that the resulting carrier could hold an amphiphilic property.

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Acknowledgements

The authors would like to thank the National Natural Science Foundation of China (No.21074049) and the Opening Foundation of State Key Laboratory of Applied Organic Chemistry (SKLAOC-2009-35) for the financial supports.

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Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, State Key Laboratory of Applied Organic Chemistry, Institute of Biochemical Engineering & Environmental Technology, Lanzhou University, Lanzhou, 730000, China

    Yanjun Cui, Xia Chen, Yanfeng Li, Xiao Liu, Lin Lei & Sunting Xuan

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  1. Yanjun Cui
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  2. Xia Chen
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  3. Yanfeng Li
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  4. Xiao Liu
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  5. Lin Lei
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  6. Sunting Xuan
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Correspondence to Yanfeng Li.

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Cui, Y., Chen, X., Li, Y. et al. Novel magnetic microspheres of P (GMA-b-HEMA): preparation, lipase immobilization and enzymatic activity in two phases. Appl Microbiol Biotechnol 95, 147–156 (2012). https://doi.org/10.1007/s00253-011-3745-x

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  • DOI: https://doi.org/10.1007/s00253-011-3745-x

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