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Immobilization and Characterization of a Thermostable Lipase

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Abstract

Lipases have found a number of commercial applications. However, thermostable lipase immobilized on nanoparticle is not extensively characterized. In this study, a recombinant thermostable lipase (designated as TtL) from Thermus thermophilus WL was expressed in Escherichia coli and immobilized onto 3-APTES-modified Fe3O4@SiO2 supermagnetic nanoparticles. Based on analyses with tricine–sodium dodecyl sulfate–polyacrylamide gel electrophoresis, X-ray diffraction, transmission electron microscopy, and vibrating sample magnetometer observation, the diameter of immobilized lipase nanoparticle was 18.4 (±2.4) nm, and its saturation magnetization value was 52.3 emu/g. The immobilized lipase could be separated from the reaction medium rapidly and easily in a magnetic field. The biochemical characterizations revealed that, comparing with the free one, the immobilized lipase exhibited better resistance to temperature, pH, metal ions, enzyme inhibitors, and detergents. The K m value for the immobilized TtL (2.56 mg/mL) was found to be lower than that of the free one (3.74 mg/mL), showing that the immobilization improved the affinity of lipase for its substrate. In addition, the immobilized TtL exhibited good reusability. It retained more than 79.5 % of its initial activity after reusing for 10 cycles. Therefore, our study presented that the possibility of the efficient reuse of the thermostable lipase immobilized on supermagnetic nanoparticles made it attractive from the viewpoint of practical application.

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Acknowledgments

This work was financially supported by China Ocean Mineral Resources R&D Association (DY125-15-E-01), the Project of State Oceanic Administration, China (201205020–3), Hi-Tech Research and Development Program of China (2012AA092103-5), and the Natural Science Foundation of Anhui Provincial University (KJ2013Z256).

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Authors and Affiliations

  1. Key Laboratory of Conservation Biology for Endangered Wildlife of Ministry of Education and College of Life Sciences, Zhejiang University, Hangzhou, 310058, The People’s Republic of China

    Chongfu Song & Xiaobo Zhang

  2. College of Chemistry and Chemical Engineering, Fuyang Teachers College, Fuyang, 236037, The People’s Republic of China

    Chongfu Song & Liangquan Sheng

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  1. Chongfu Song
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  2. Liangquan Sheng
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  3. Xiaobo Zhang
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Correspondence to Liangquan Sheng or Xiaobo Zhang.

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Song, C., Sheng, L. & Zhang, X. Immobilization and Characterization of a Thermostable Lipase. Mar Biotechnol 15, 659–667 (2013). https://doi.org/10.1007/s10126-013-9515-2

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