3W25

The high-resolution crystal structure of TsXylA, intracellular xylanase from /Thermoanaerobacterium saccharolyticum JW/SL-YS485/: the complex of the E146A mutant with xylobiose

Summary for 3W25

• Entry DOI: 10.2210/pdb3w25/pdb
• Related: 3W24 3W26 3W27 3W28 3W29 3W2B
• Related PRD ID: PRD_900116
• Descriptor: Glycoside hydrolase family 10, beta-D-xylopyranose-(1-4)-beta-D-xylopyranose (3 entities in total)
• Functional Keywords: glycoside hydrolase, xylanase, thermophilic, xylobiose, hydrolase
• Biological source: Thermoanaerobacterium saccharolyticum
• Total number of polymer chains: 1
• Total formula weight: 38287.26

Authors

Han, X.,Gao, J.,Shang, N.,Huang, C.-H.,Ko, T.-P.,Zhu, Z.,Wiegel, J.,Shao, W.,Guo, R.-T. (deposition date: 2012-11-27, release date: 2013-04-03, Last modification date: 2023-11-08)

Primary citation

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.,Ma, Y.
Structural and functional analyses of catalytic domain of GH10 xylanase from Thermoanaerobacterium saccharolyticum JW/SL-YS485
Proteins, 81:1256-1265, 2013

PubMed 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.

PubMed: 23508990
DOI: 10.1002/prot.24286

Experimental method

X-RAY DIFFRACTION (1.32 Å)