Get access

Structural and functional analyses of catalytic domain of GH10 xylanase from Thermoanaerobacterium saccharolyticum JW/SL-YS485

Authors

  • Xu Han,

    1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
    • Xu Han, Jian Gao, and Na Shang contributed equally to this work.

  • Jian Gao,

    1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
    • Xu Han, Jian Gao, and Na Shang contributed equally to this work.

  • Na Shang,

    1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
    • Xu Han, Jian Gao, and Na Shang contributed equally to this work.

  • Chun-Hsiang Huang,

    1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
  • Tzu-Ping Ko,

    1. Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan
    Search for more papers by this author
  • Chun-Chi Chen,

    1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
    Search for more papers by this author
  • Hsiu-Chien Chan,

    1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
  • Ya-Shan Cheng,

    1. AsiaPac Biotechnology Co., Ltd., Dongguan, China
    Search for more papers by this author
  • Zhen Zhu,

    1. Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
  • Juergen Wiegel,

    1. Department of Microbiology, University of Georgia, Athens, Georgia
    Search for more papers by this author
  • Wenhua Luo,

    Corresponding author
    • College of Food Science, South China Agricultural University, Guangzhou, China
    Search for more papers by this author
  • Rey-Ting Guo,

    Corresponding author
    • Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author
  • Yanhe Ma

    Corresponding author
    • Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, China
    Search for more papers by this author

Yanhe Ma, 32 XiQiDao, Tianjin Airport Economic Park, Tianjin 300308, China. E-mail: ma_yh@tib.cas.cn or Rey-Ting Guo, 32 XiQiDao, Tianjin Airport Economic Park, Tianjin 300308, China. E-mail: guo_rt@tib.cas.cn or Wenhua Luo, No. 3 Gongye North Road, Songshan Lake Scientific Industrial Park, Dongguan, Guangdong Province, China. E-mail: wenhualuo@126.com

ABSTRACT

Xylanases are capable of decomposing xylans, the major components in plant cell wall, and releasing the constituent sugars for further applications. Because xylanase is widely used in various manufacturing processes, high specific activity, and thermostability are desirable. Here, the wild-type and mutant (E146A and E251A) catalytic domain of xylanase from Thermoanaerobacterium saccharolyticum JW/SL-YS485 (TsXylA) were expressed in Escherichia coli and purified subsequently. The recombinant protein showed optimal temperature and pH of 75°C and 6.5, respectively, and it remained fully active even after heat treatment at 75°C for 1 h. Furthermore, the crystal structures of apo-form wild-type TsXylA and the xylobiose-, xylotriose-, and xylotetraose-bound E146A and E251A mutants were solved by X-ray diffraction to high resolution (1.32–1.66 Å). The protein forms a classic (β/α)8 folding of typical GH10 xylanases. The ligands in substrate-binding groove as well as the interactions between sugars and active-site residues were clearly elucidated by analyzing the complex structures. According to the structural analyses, TsXylA utilizes a double displacement catalytic machinery to carry out the enzymatic reactions. In conclusion, TsXylA is effective under industrially favored conditions, and our findings provide fundamental knowledge which may contribute to further enhancement of the enzyme performance through molecular engineering. Proteins 2013; 81:1256–1265. © 2013 Wiley Periodicals, Inc.

Get access to the full text of this article

Ancillary