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Secretory expression and characterization of a novel thermo-stable, salt-tolerant endo-1,4-β-mannanase of Bacillus subtilis WD23 by Pichia pastoris

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Abstract

Endo-1,4-β-mannanases have a wide range of potential industrial applications. It was mainly used for the production of functional oligosaccharides in the food industry. In order to increase the production of endo-1,4-β-mannanases, inducible and constitutive expression of a novel endo-1,4-β-mannanase from Bacillus subtilis WD23 in Pichia pastoris GS115 was performed and the recombinant enzyme was characterized. The recombinant mannanase gene was successfully expressed in a fully active form in P. pastoris GS115, and the activity of recombinant mannanase reached to 5,156.742 U/mL, which was higher than those from other recombinant system. Optimal purified enzyme activity was detected at pH 6.3 and 70 °C, and the purified enzyme displayed broad temperature stability (40–70 °C). 150 mg/mL NaCl could still improve the activity of the enzyme. Flight mass spectrometry determination confirmed that recombinant proteins are mannanase.

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Acknowledgments

This study was supported by the state forestry administration project 948 (2012-4-03), National Natural Science Foundation (No.31170553, 30671702, 30170775), excellent creative talents supporting program of Heilongjiang University of Chinese Medicine (No. 2012RCQ13), and the Fundamental Research Funds for the Central Universities (No. DL12CA08).

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None.

Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. College of Life Sciences, Northeast Forestry University, Harbin, 150040, China

    Huiling Li, Zuyan Liu, Chunlei Wang & Min Zhao

  2. College of Pharmacy, Heilongjiang University of Chinese Medicine, Harbin, 150040, China

    Huiling Li

  3. Agricultural College, Yanbian University, Yanji, 133002, China

    Shichen Huang

Authors
  1. Huiling Li
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  2. Zuyan Liu
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  3. Chunlei Wang
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  4. Shichen Huang
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  5. Min Zhao
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Corresponding author

Correspondence to Min Zhao.

Additional information

Huiling Li and Zuyan Liu have contributed equally to this article.

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Li, H., Liu, Z., Wang, C. et al. Secretory expression and characterization of a novel thermo-stable, salt-tolerant endo-1,4-β-mannanase of Bacillus subtilis WD23 by Pichia pastoris . Eur Food Res Technol 240, 671–677 (2015). https://doi.org/10.1007/s00217-014-2369-7

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  • DOI: https://doi.org/10.1007/s00217-014-2369-7

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