| Content Provider |
World Health Organization (WHO)-Global Index Medicus |
| Author |
Yang, Barbara Liang, Po-huang Ho, Meng-chiao Yuan, Shuo-fu Gao, Jian Li, Qian Huang, Chun-hsiang Guo, Rey-ting Hsieh, Han-yu Lee, Hsiao-lin Lin, Wen-ling Chang, Chih-kang Wu, Tzu-hui |
| Description |
Author Affiliation: Yuan SF ( the Institute of Biochemical Sciences, and.); Wu TH ( the Institute of Biotechnology, National Taiwan University, Taipei 10617, Taiwan and.); Lee HL ( From the Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.); Hsieh HY ( the Institute of Biochemical Sciences, and.); Lin WL ( From the Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.); Yang B ( the Institute of Biochemical Sciences, and.); Chang CK ( From the Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.); Li Q ( the Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.); Gao J ( the Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.); Huang CH ( the Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.); Ho MC ( the Institute of Biochemical Sciences, and From the Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan, joeho@gate.sinica.edu.tw.); Guo RT ( the Industrial Enzymes National Engineering Laboratory, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China guo_rt@tib.cas.cn.); Liang PH ( the Institute of Biochemical Sciences, and From the Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan, phliang@gate.sinica.edu.tw.) |
| Abstract |
We expressed an active form of CtCel5E (a bifunctional cellulase/xylanase from Clostridium thermocellum), performed biochemical characterization, and determined its apo- and ligand-bound crystal structures. From the structures, Asn-93, His-168, His-169, Asn-208, Trp-347, and Asn-349 were shown to provide hydrogen-bonding/hydrophobic interactions with both ligands. Compared with the structures of TmCel5A, a bifunctional cellulase/mannanase homolog from Thermotoga maritima, a flexible loop region in CtCel5E is the key for discriminating substrates. Moreover, site-directed mutagenesis data confirmed that His-168 is essential for xylanase activity, and His-169 is more important for xylanase activity, whereas Asn-93, Asn-208, Tyr-270, Trp-347, and Asn-349 are critical for both activities. In contrast, F267A improves enzyme activities. |
| ISSN |
00219258 |
| e-ISSN |
1083351X |
| Journal |
Journal of Biological Chemistry |
| Issue Number |
9 |
| Volume Number |
290 |
| Language |
English |
| Publisher |
American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date |
2015-02-27 |
| Publisher Place |
United States |
| Access Restriction |
Open |
| Subject Keyword |
Bacterial Proteins Chemistry Cellulase Clostridium Thermocellum Enzymology Endo-1,4-beta Xylanases Protein Structure, Tertiary Amino Acid Sequence Amino Acids Genetics Metabolism Binding Sites Catalytic Domain Cellobiose Crystallography, X-Ray Disaccharides Enzyme Assays Kinetics Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Binding Recombinant Proteins Sequence Homology, Amino Acid Substrate Specificity Thermotoga Maritima Research Support, Non-U.S. Gov't Biochemistry Molecular Biology |
| Content Type |
Text |
| Resource Type |
Article |
| Subject |
Cell Biology Biochemistry Molecular Biology |
