Structural Insights into Rice BGlu1 β-Glucosidase Oligosaccharide Hydrolysis and Transglycosylation

Watchalee Chuenchor, Salila Pengthaisong, Robert C. Robinson, Jirundon Yuvaniyama, Worrapoj Oonanant, David R. Bevan, Asim Esen, Chun Jung Chen, Rodjana Opassiri, Jisnuson Svasti, James R Ketudat Cairns

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

The structures of rice BGlu1 β-glucosidase, a plant β-glucosidase active in hydrolyzing cell wall-derived oligosaccharides, and its covalent intermediate with 2-deoxy-2-fluoroglucoside have been solved at 2.2 Å and 1.55 Å resolution, respectively. The structures were similar to the known structures of other glycosyl hydrolase family 1 (GH1) β-glucosidases, but showed several differences in the loops around the active site, which lead to an open active site with a narrow slot at the bottom, compatible with the hydrolysis of long β-1,4-linked oligosaccharides. Though this active site structure is somewhat similar to that of the Paenibacillus polymyxa β-glucosidase B, which hydrolyzes similar oligosaccharides, molecular docking studies indicate that the residues interacting with the substrate beyond the conserved -1 site are completely different, reflecting the independent evolution of plant and microbial GH1 exo-β-glucanase/β-glucosidases. The complex with the 2-fluoroglucoside included a glycerol molecule, which appears to be in a position to make a nucleophilic attack on the anomeric carbon in a transglycosylation reaction. The coordination of the hydroxyl groups suggests that sugars are positioned as acceptors for transglycosylation by their interactions with E176, the catalytic acid/base, and Y131, which is conserved in barley BGQ60/β-II β-glucosidase, that has oligosaccharide hydrolysis and transglycosylation activity similar to rice BGlu1. As the rice and barley enzymes have different preferences for cellobiose and cellotriose, residues that appeared to interact with docked oligosaccharides were mutated to those of the barley enzyme to see if the relative activities of rice BGlu1 toward these substrates could be changed to those of BGQ60. Although no single residue appeared to be responsible for these differences, I179, N190 and N245 did appear to interact with the substrates.

Original languageEnglish
Pages (from-to)1200-1215
Number of pages16
JournalJournal of Molecular Biology
Volume377
Issue number4
DOIs
Publication statusPublished - Apr 4 2008
Externally publishedYes

Fingerprint

Glucosidases
Oligosaccharides
Hydrolysis
Hordeum
Catalytic Domain
Hydrolases
Cellobiose
Enzymes
Hydroxyl Radical
Glycerol
Cell Wall
Carbon
Oryza
Acids

Keywords

  • β-glucosidase
  • cellooligosaccharides
  • enzyme-substrate interactions
  • structure function relationship
  • X-ray crystallography

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Chuenchor, W., Pengthaisong, S., Robinson, R. C., Yuvaniyama, J., Oonanant, W., Bevan, D. R., ... Cairns, J. R. K. (2008). Structural Insights into Rice BGlu1 β-Glucosidase Oligosaccharide Hydrolysis and Transglycosylation. Journal of Molecular Biology, 377(4), 1200-1215. https://doi.org/10.1016/j.jmb.2008.01.076

Structural Insights into Rice BGlu1 β-Glucosidase Oligosaccharide Hydrolysis and Transglycosylation. / Chuenchor, Watchalee; Pengthaisong, Salila; Robinson, Robert C.; Yuvaniyama, Jirundon; Oonanant, Worrapoj; Bevan, David R.; Esen, Asim; Chen, Chun Jung; Opassiri, Rodjana; Svasti, Jisnuson; Cairns, James R Ketudat.

In: Journal of Molecular Biology, Vol. 377, No. 4, 04.04.2008, p. 1200-1215.

Research output: Contribution to journalArticle

Chuenchor, W, Pengthaisong, S, Robinson, RC, Yuvaniyama, J, Oonanant, W, Bevan, DR, Esen, A, Chen, CJ, Opassiri, R, Svasti, J & Cairns, JRK 2008, 'Structural Insights into Rice BGlu1 β-Glucosidase Oligosaccharide Hydrolysis and Transglycosylation', Journal of Molecular Biology, vol. 377, no. 4, pp. 1200-1215. https://doi.org/10.1016/j.jmb.2008.01.076
Chuenchor, Watchalee ; Pengthaisong, Salila ; Robinson, Robert C. ; Yuvaniyama, Jirundon ; Oonanant, Worrapoj ; Bevan, David R. ; Esen, Asim ; Chen, Chun Jung ; Opassiri, Rodjana ; Svasti, Jisnuson ; Cairns, James R Ketudat. / Structural Insights into Rice BGlu1 β-Glucosidase Oligosaccharide Hydrolysis and Transglycosylation. In: Journal of Molecular Biology. 2008 ; Vol. 377, No. 4. pp. 1200-1215.
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