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Expression and Characterization of Recombinant Sucrose Phosphorylase

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Abstract

SPase is widely used in the food, cosmetics, and pharmaceutical industries. Previously, a SPase gene was cloned from Bifidobacterium longum JCM1217 and constructed into Escherichia coli BL21. In this paper, its expression conditions were optimized. The results showed that several induction factors determined the expression efficiency of SPase. The initial cell density, IPTG concentration, and induction time and temperature significantly (p < 0.01) affected the total protein content and activity of expressed SPase. The highest expression efficiency was obtained at an initial cell density of OD600 = 0.5, with 0.05 mM IPTG, followed by shaking at 180 rpm and incubation at 30 °C for 15 h. The purified SPase had a specific activity of 122.1 U/mg, which was raised by 1.85 -fold more than that before optimization, and its recovery yield was 86%. Furthermore, SPase also showed higher thermostability. The results of this study provide essential information for the industrial production of SPase.

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Abbreviations

SPase:

Sucrose phosphorylase

IPTG:

Isopropyl-β-d-thiogalactoside

Glc 1-P:

α-d-Glucose 1-phosphate

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Acknowledgements

This work was supported by the program for International S & T Cooperation Projects of Shenyang (F16-219-6-00), the research Grants of Shenyang Kay Laboratory (F17-158-1-00), and the research foundation of Shenyang Agricultural University.

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Authors and Affiliations

  1. College of Food Science, Shenyang Agricultural University, Shenyang, 110866, China

    Hui Zhang, Tuoping Li & Suhong Li

  2. Shenyang Women’s and Children’s Hospital, Shenyang, 110011, China

    Xiao Sun & Wenjie Li

  3. National Food Research Institute, Tsukuba, 305-8642, Japan

    Motomitsu Kitaoka

Authors
  1. Hui Zhang
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  2. Xiao Sun
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  3. Wenjie Li
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  4. Tuoping Li
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  5. Suhong Li
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  6. Motomitsu Kitaoka
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Correspondence to Tuoping Li.

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Zhang, H., Sun, X., Li, W. et al. Expression and Characterization of Recombinant Sucrose Phosphorylase. Protein J 37, 93–100 (2018). https://doi.org/10.1007/s10930-017-9754-6

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  • DOI: https://doi.org/10.1007/s10930-017-9754-6

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