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Characterization of a neutral recombinant xylanase from Thermoactinospora rubra YIM 77501T

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Abstract

A xylanase gene (TrXyn10) from Thermoactinospora rubra YIM 77501T was cloned and expressed in Escherichia coli. The amino acid sequence displayed 78% homology with Microbispora mesophila xylanase (WP_062413927.1). The recombinant xylanase (TrXyn10), with MW 46.1 kDa, could hydrolyse beechwood, birchwood and oatspelt xylan. Based on the sequence, enzymatic properties and tertiary structure of the protein, TrXyn10 belongs to glycoside hydrolase family 10 (GH10). The optimal pH and temperature for the recombinant enzyme were determined to be 7.0 and 55 °C, respectively. TrXyn10 was stable over a wide pH range, and it retained more than 45% of the total activity at pH 6.0–12.0 for 12 h. In addition, the activity was greatly promoted, by approximately 200% of the initial activity, after incubation at pH 6.0 and 7.0 for 12 h. Based on enzymatic properties and product analysis, we showed that TrXyn10 is a neutral endoxylanase.

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Acknowledgements

This work was supported by the Key Project of International Cooperation of Ministry of Science & Technology (MOST) (No. 2013DFA31980), Science and technology infrastructure work project (No. 2015FY110100), Natural Science Foundation of China (No. 3150004), Yunnan Provincial Natural Science Foundation (2013FA004) and Scientific Research Fund of Xinxiang Medical University (2013QN126). W-J Li was also supported by Guangdong Province Higher Vocational Colleges & Schools Pearl River Scholar Funded Scheme (2014).

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    Authors and Affiliations

    1. Yunnan Institute of Microbiology, Yunnan University, 650091, Kunming, People’s Republic of China

      Yi-Rui Yin, Qing-Wen Hu, Hong Ming, Xiao-Yang Zhi & Wen-Jun Li

    2. State Key Laboratory of Biocontrol and Guangdong Provincial Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-Sen University, 510275, Guangzhou, People’s Republic of China

      Wen-Dong Xian, En-Min Zhou, Min Xiao & Wen-Jun Li

    3. Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, and School of Life Sciences, Fujian Agriculture and Forestry University, 350002, Fuzhou, People’s Republic of China

      Feng Zhang

    4. College of Life Sciences and Technology, Xinxiang Medical University, 453003, Xinxiang, People’s Republic of China

      Hong Ming

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    1. Yi-Rui Yin
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    Correspondence to Xiao-Yang Zhi or Wen-Jun Li.

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    Yi-Rui Yin and Qing-Wen Hu have contributed equally to this work.

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    Yin, YR., Hu, QW., Xian, WD. et al. Characterization of a neutral recombinant xylanase from Thermoactinospora rubra YIM 77501T . Antonie van Leeuwenhoek 110, 429–436 (2017). https://doi.org/10.1007/s10482-016-0798-y

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