9vmk | pdb_00009vmk
Crystal structure of TeGH116 from Thermosynechococcus elongatus with glucose
Structural highlights
FunctionPublication Abstract from PubMedbeta-Glucosidases play essential roles in nature as well as in industrial applications. Glycoside hydrolase family 116 (GH116) is a relatively sparsely characterized family of beta-glucosidases. We describe the functional and structural characterization of Thermosynechococcus elongatus BP-1 TeGH116, the one beta-glucosidase in this model cyanobacterium. TeGH116 was expressed in Escherichia coli and purified by heating, immobilized metal affinity chromatography, and size exclusion chromatography. TeGH116 hydrolyzes p-nitrophenyl beta-d-glucopyranoside most rapidly at pH 5.5 and 55 degrees C, and other p-nitrophenyl glycosides 3% or less of this rate. TeGH116 also hydrolyzes natural phenolic glucosides and beta-1,3- and beta-1,4-linked gluco-oligosaccharides. Kinetic analysis suggests that TeGH116 binds three glucosyl residues in these oligosaccharides. TeGH116 is inhibited by glucose but is only weakly inhibited by delta-gluconolactone. The X-ray crystallographic structures of TeGH116 and its complex with glucose were similar to those of Thermoanaerobacterium xylanolyticum TxGH116. However, a long loop extending from the noncatalytic N-terminal domain helps form the mouth of the TeGH116 active site, making it narrower. TxGH116 was found to hydrolyze barley beta-glucan, laminarin, and lichenan, but not carboxymethylcellulose, while TeGH116 hydrolyzed the same polysaccharides at relatively lower rates. Deletion of the loop in TeGH116 resulted in poor stability and low activity, but higher relative activity on laminarin compared to p-nitrophenyl beta-d-glucoside. The narrowed active site and lower activity on large substrates highlight the contribution of N-terminal domain loops to the substrate specificity of GH116 enzymes. The structure of Thermosynechococcus elongatus glycoside hydrolase family 116 beta-glucosidase shows the role of the noncatalytic N-terminal domain in controlling substrate specificity.,Beagbandee C, Charoenwattanasatien R, Pengthaisong S, Kurisu G, Ketudat Cairns JR Int J Biol Macromol. 2025 Dec;332(Pt 1):148566. doi: , 10.1016/j.ijbiomac.2025.148566. Epub 2025 Oct 27. PMID:41161445[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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