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Expression and biochemical characterization of a novel type I pullulanase from Bacillus megaterium

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Abstract

Objectives

To identify novel pullulanases from microorganisms and to investigate their biochemical characterizations.

Results

A novel pullulanase gene (BmPul) from Bacillus megaterium WW1210 was cloned and heterologously expressed in Escherichia coli. The gene has an ORF of 2814 bp encoding 937 amino acids. The recombinant pullulanase (BmPul) was purified to homogeneity and biochemically characterized. BmPul has an MW of approx. 112 kDa as indicated by SDS-PAGE. Optimum conditions were at 55 °C and pH 6.5. The enzyme was stable below 40 °C and from pH 6.5−8.5. The Km values of BmPul towards pullulan and amylopectin were 3.3 and 3.6 mg/ml, respectively. BmPul hydrolyzed pullulan to yield mainly maltotriose, indicating that it should be a type I pullulanase.

Conclusions

A novel type I pullulanase from Bacillus megaterium was identified, heterologously expressed and biochemically characterized. Its properties makes this enzyme as a good candidate for the food industry.

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Acknowledgements

This work was financially supported by the National Science Fund for Distinguished Young Scholars (No. 31325021) and the Program for Changjiang Scholars (No. T2014055).

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Authors and Affiliations

  1. Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing, 100083, China

    Shaoqing Yang, Jingjing Liu & Zhengqiang Jiang

  2. Beijing Advanced Innovation Center for Food Nutrition and Human Health, Bioresource Utilization Laboratory, College of Engineering, China Agricultural University, Beijing, 100083, China

    Qiaojuan Yan & Qingdan Bao

Authors
  1. Shaoqing Yang
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  2. Qiaojuan Yan
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  3. Qingdan Bao
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  4. Jingjing Liu
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  5. Zhengqiang Jiang
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Corresponding author

Correspondence to Zhengqiang Jiang.

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Yang, S., Yan, Q., Bao, Q. et al. Expression and biochemical characterization of a novel type I pullulanase from Bacillus megaterium . Biotechnol Lett 39, 397–405 (2017). https://doi.org/10.1007/s10529-016-2255-4

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  • DOI: https://doi.org/10.1007/s10529-016-2255-4

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