Xu Han, Jian Gao, and Na Shang contributed equally to this work.
Structural and functional analyses of catalytic domain of GH10 xylanase from Thermoanaerobacterium saccharolyticum JW/SL-YS485
Article first published online: 20 APR 2013
Copyright © 2013 Wiley Periodicals, Inc.
Proteins: Structure, Function, and Bioinformatics
Volume 81, Issue 7, pages 1256–1265, July 2013
How to Cite
Han, X., Gao, J., Shang, N., Huang, C.-H., Ko, T.-P., Chen, C.-C., Chan, H.-C., Cheng, Y.-S., Zhu, Z., Wiegel, J., Luo, W., Guo, R.-T. and Ma, Y. (2013), Structural and functional analyses of catalytic domain of GH10 xylanase from Thermoanaerobacterium saccharolyticum JW/SL-YS485. Proteins, 81: 1256–1265. doi: 10.1002/prot.24286
- Issue published online: 17 JUN 2013
- Article first published online: 20 APR 2013
- Accepted manuscript online: 18 MAR 2013 10:44AM EST
- Manuscript Accepted: 5 MAR 2013
- Manuscript Revised: 1 MAR 2013
- Manuscript Received: 3 JAN 2013
- National Basic Research Program of China . Grant Number: 2011CBA00800
- National High Technology Research and Development Program of China . Grant Number: 2012AA022200
- Chinese Academy of Sciences . Grant Number: 2011B09030042
- Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
- glycoside hydrolase;
- crystal structure;
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.