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Covalent Immobilization and Characterization of a Novel Pullulanase from Fontibacillus sp. Strain DSHK 107 onto Florisil® and Nano-silica for Pullulan Hydrolysis

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Abstract

A novel pullulanase partially purified from Fontibacillus sp. was covalently immobilized on Florisil® and nano-silica through both glutaraldehyde and (3-glycidyloxypropyl)trimethoxysilane spacer arms. The pullulanase immobilized on Florisil® and nano-silica through glutaraldehyde spacer arm showed 85 and 190 % activity of its free form, respectively, whereas no activity was observed when it was immobilized on the same supports through (3-glycidyloxypropyl)trimethoxysilane spacer arm. The maximum working pHs of both the immobilized pullulanases on Florisil® and nano-silica through glutaraldehyde spacer arm were determined as 5.0; however, the maximum working pH of the free pullulanase was pH 6.0. The maximum temperatures of all the pullulanase preparations were determined as 35 °C. The apparent K m values were 1.49, 1.54, and 0.59 mg/mL pullunan, respectively, for the free and immobilized pullulanases on Florisil® and nano-silica. The corresponding apparent V max values were 0.59, 1.53, and 1.57 U mg prot.−1 min.−1. Thermal stability of pullulanases immobilized on Florisil® and nano-silica was enhanced 6.5- and 15.6-folds, respectively at 35 °C and 6.6- and 16.0-folds, respectively, at 50 °C. The pullulanases immobilized on Florisil® and nano-silica protected 71 and 90 % of their initial activities after 10 reuses.

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Acknowledgments

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Vocational School of Imamoglu, University of Cukurova, Adana, Turkey

    Dilek Alagöz

  2. Vocational School of Ceyhan, University of Cukurova, Adana, Turkey

    Deniz Yildirim

  3. Faculty of Sciences and Letters, Department of Biology, University of Cukurova, Adana, Turkey

    Hatice Korkmaz Güvenmez & Damla Sihay

  4. Faculty of Sciences and Letters, Department of Chemistry, University of Cukurova, Adana, Turkey

    S. Seyhan Tükel

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  1. Dilek Alagöz
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Correspondence to Dilek Alagöz.

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Alagöz, D., Yildirim, D., Güvenmez, H.K. et al. Covalent Immobilization and Characterization of a Novel Pullulanase from Fontibacillus sp. Strain DSHK 107 onto Florisil® and Nano-silica for Pullulan Hydrolysis. Appl Biochem Biotechnol 179, 1262–1274 (2016). https://doi.org/10.1007/s12010-016-2063-2

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  • DOI: https://doi.org/10.1007/s12010-016-2063-2

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