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Updated: Mar 25, 2026
09:27
Rapid One-step Enzymatic Synthesis and All-aqueous Purification of Trehalose Analogues
Published on: February 17, 2017
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Bacillus licheniformis trehalose-6-phosphate hydrolase structures suggest keys to substrate specificity
Min Guan Lin1, Meng Chun Chi2, Vankadari Naveen3
1Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu 300, Taiwan.
Acta Crystallographica. Section D, Structural Biology
|February 20, 2016
Summary
Trehalose-6-phosphate hydrolase (TreA) is a glycoside hydrolase. Structural analysis reveals unique active site features and positively charged residues critical for substrate discrimination in Bacillus licheniformis TreA.
Area of Science:
- Biochemistry
- Structural Biology
- Enzymology
Background:
- Trehalose-6-phosphate hydrolase (TreA) is a glycoside hydrolase family 13 (GH13) enzyme.
- TreA catalyzes the hydrolysis of trehalose 6-phosphate (T6P) into glucose and glucose 6-phosphate, feeding into the glycolytic pathway.
Purpose of the Study:
- To elucidate the structural basis of TreA's function and substrate specificity.
- To investigate the role of specific active site residues in the enzymatic activity of Bacillus licheniformis TreA (BlTreA).
Main Methods:
- X-ray crystallography was used to determine the structures of BlTreA and its R201Q mutant complexed with p-nitrophenyl-α-D-glucopyranoside (pPNG) at high resolution.
- Site-directed mutagenesis was employed to investigate the function of key active site residues.
Main Results:
- The overall structure of BlTreA is similar to other GH13 enzymes, but features a unique loop conformation in the catalytic site.
- The active site exhibits a distinct positive surface potential due to basic residues (His281, His282, Lys284, Lys292).
- Mutations in these residues significantly reduced enzymatic activity, highlighting their importance.
Conclusions:
- The unique active site structure and positive charge distribution are crucial for BlTreA's catalytic activity.
- The (281)HHLK(284) motif and Lys292 are critical for substrate discrimination by BlTreA.
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