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Purification, Identification, and Characterization of a Glycoside Hydrolase Family 11-Xylanase with High Activity from Aspergillus niger VTCC 017

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Abstract

Xylanases (EC 3.2.1.8) have been considered as a potential green solution for the sustainable development of a wide range of industries including pulp and paper, food and beverages, animal feed, pharmaceuticals, and biofuels because they are the key enzymes that degrade the xylosidic linkages of xylan, the major component of the second most abundant raw material worldwide. Therefore, there is a critical need for the industrialized xylanases which must have high specific activity, be tolerant to organic solvent or detergent and be active during a wide range of conditions, such as high temperature and pH. In this study, an extracellular xylanase was purified from the culture broth of Aspergillus niger VTCC 017 for primary structure determination and properties characterization. The successive steps of purification comprised centrifugation, Sephadex G-100 filtration, and DEAE-Sephadex chromatography. The purified xylanase (specific activity reached 6596.79 UI/mg protein) was a monomer with a molecular weight of 37 kDa estimating from SDS electrophoresis. The results of LC/MS suggested that the purified protein is indeed an endo-1,4-β-d-xylanase. The purified xylanase showed the optimal temperature of 55 °C, and pH 6.5 with a stable xylanolytic activity within the temperature range of 45–50 °C, and within the pH range of 5.0–8.0. Most divalent metal cations including Zn2+, Fe2+, Mg2+, Cu2+, Mn2+ showed some inhibition of xylanase activity while the monovalent metal cations such as K+ and Ag+ exhibited slight stimulating effects on the enzyme activity. The introduction of 10–30% different organic solvents (n-butanol, acetone, isopropanol) and several detergents (Triton X-100, Tween 20, and SDS) slightly reduced the enzyme activity. Moreover, the purified xylanase seemed to be tolerant to methanol and ethanol and was even stimulated by Tween 80. Overall, with these distinctive properties, the putative xylanase could be a successful candidate for numerous industrial uses.

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Acknowledgements

This study was supported by the National Foundation for Science and Technology Development Vietnam (Nafosted), project 106.02−2018.347” Engineering of recombinant Aspergillus niger to produce highly active xylanase for functional foods industry” 2019−2021.

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Authors and Affiliations

  1. Department of Biochemistry, Hanoi University of Pharmacy, Hanoi, Vietnam

    Dao Thi Mai Anh

  2. Institute of Biotechnology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet Road, Caugiay District, 10600, Hanoi, Vietnam

    Nguyen Tien Cuong & Do Thi Tuyen

  3. Graduate University of Science and Technology, Vietnam Academy of Science and Technology, Hanoi, Vietnam

    Nguyen Thi Trung & Do Thi Tuyen

  4. Tay Nguyen University, Buon Ma Thuot, Dak Lak, Vietnam

    Nguyen Phuong Dai Nguyen

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  1. Dao Thi Mai Anh
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  2. Nguyen Tien Cuong
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  4. Nguyen Phuong Dai Nguyen
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  5. Do Thi Tuyen
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Contributions

DTT and DTMA designed the experimental setup and manuscript preparation. NTC and DTMA performed experiments of purification of xylanase. NTT and NPDN performed and characterized xylanase. NTT and DTT performed and evaluated data analysis of LC/MS analysis of purified xylanase. DTT initiated the project, read and approved the final manuscript. All authors read and approved the final manuscript.

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Correspondence to Do Thi Tuyen.

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Dao, T.M.A., Cuong, N.T., Nguyen, T.T. et al. Purification, Identification, and Characterization of a Glycoside Hydrolase Family 11-Xylanase with High Activity from Aspergillus niger VTCC 017. Mol Biotechnol 64, 187–198 (2022). https://doi.org/10.1007/s12033-021-00395-8

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  • DOI: https://doi.org/10.1007/s12033-021-00395-8

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