1. Structural analyses and yeast production of the β-1,3-1,4-glucanase catalytic module encoded by the licB gene of Clostridium thermocellum.
- Author
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Chen CC, Huang JW, Zhao P, Ko TP, Huang CH, Chan HC, Huang Z, Liu W, Cheng YS, Liu JR, and Guo RT
- Subjects
- Amino Acid Sequence, Bacterial Proteins genetics, Catalytic Domain, Clostridium thermocellum genetics, Crystallography, X-Ray, Enzyme Stability, Escherichia coli enzymology, Escherichia coli genetics, Genes, Bacterial, Glycoside Hydrolases genetics, Industrial Microbiology, Models, Molecular, Molecular Sequence Data, Pichia enzymology, Pichia genetics, Protein Conformation, Protein Interaction Domains and Motifs, Protein Structure, Secondary, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Recombinant Proteins genetics, Sequence Homology, Amino Acid, Static Electricity, Structural Homology, Protein, Bacterial Proteins biosynthesis, Bacterial Proteins chemistry, Clostridium thermocellum enzymology, Glycoside Hydrolases biosynthesis, Glycoside Hydrolases chemistry
- Abstract
A thermophilic glycoside hydrolase family 16 (GH16) β-1,3-1,4-glucanase from Clostridium thermocellum (CtLic16A) holds great potentials in industrial applications due to its high specific activity and outstanding thermostability. In order to understand its molecular machinery, the crystal structure of CtLic16A was determined to 1.95Å resolution. The enzyme folds into a classic GH16 β-jellyroll architecture which consists of two β-sheets atop each other, with the substrate-binding cleft lying on the concave side of the inner β-sheet. Two Bis-Tris propane molecules were found in the positive and negative substrate binding sites. Structural analysis suggests that the major differences between the CtLic16A and other GH16 β-1,3-1,4-glucanase structures occur at the protein exterior. Furthermore, the high catalytic efficacy and thermal profile of the CtLic16A are preserved in the enzyme produced in Pichia pastoris, encouraging its further commercial applications., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
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