New to the CAZy classification? Read this first.
Want to learn more about CAZypedia? Read the CAZypedia 10th anniversary article in Glycobiology.
Glycoside Hydrolase Family 17
Responsible Curator as essentially complete. CAZypedia is a living document, so further improvement of this page is still possible. If you would like to suggest an addition or correction, please contact the page's Responsible Curator directly by e-mail.
|Glycoside Hydrolase Family GH17|
|Active site residues||known|
|CAZy DB link|
The family GH17 glycoside hydrolases are clan GH-A enzymes from bacteria, fungi and plants, and include two major groups of enzymes with related but distinct substrate specificities, namely 1,3-β-D-glucan endohydrolases (EC 184.108.40.206) and 1,3;1,4-β-D-glucan endohydrolases (EC 220.127.116.11). A 1,3-β-D-glucan exohydrolase (EC 18.104.22.168) is also classified in this family. The family GH17 enzymes have quite distinct amino acid sequences and 3D structures compared with the 1,3-β-D-glucan endohydrolases and 1,3;1,4-β-D-glucan endohydrolases that have similar substrate specificities but are classified in families GH16, GH55, GH64 and GH81.
The family GH17 1,3-β-D-glucan endohydrolases hydrolyse internal 1,3-β-D-glucosidic linkages in polysaccharides, but usually require a region of contiguous unbranched, un-substituted 1,3-β-D-glucosyl residues for activity. The enzymes release 1,3-β-D-oligoglucosides of DP 2-5 as their major products. Because the 1,3-β-D-glucan endohydrolases require a region of contiguous unbranched, unsubstituted 1,3-β-D-glucosyl residues for activity, they are unable to hydrolyse the single 1,3-β-D-glucosidic linkages in 1,3;1,4-β-D-glucans from the Poaceae, but they will hydrolyse 1,3-β-D-glucosidic linkages in fungal 1,3;1,6-β-D-glucans, provided an appropriate region of contiguous un-substituted 1,3-β-D-glucosyl residues is available. The family GH17 1,3;1,4-β-D-glucan endohydrolases (EC 22.214.171.124) hydrolyse 1,4-β-D-glucosidic linkages, but only 1,3;1,4-β-D-glucans in which the glucosyl residue involved in the glycosidic linkage cleaved is substituted at the C(0)3 position, that is, where the 1,4-β-D-glucosidic linkages are located on the reducing end side of 1,3-β-D-glucosyl residues.
Reaction products released are mainly 1,3;1,4-β-D-tri- and tetrasaccharides (G4G3Gred and G4G4G3Gred), but they also release higher oligosaccharides of up to 10 or more contiguous 1,4-β-D-glucosyl residues with a single reducing terminal 1,3-β-D-glucosyl residue (e.g. G4G4G4G4G4G4G3Gred). These longer oligosaccharides originate from the longer regions of adjacent 1,4-linkages that account for approximately 10% by weight of 1,3;1,4-β-D-glucans in cell walls of the Poaceae .
Kinetics and Mechanism
The stereochemistry of the reaction has been determined experimentally and catalysis by GH17 enzymes occurs via a classical Koshland retaining mechanism with the β-anomeric configuration of the released oligosaccharide being retained . Detailed kinetic analyses are available for three purified barley 1,3-β-D-glucan endohydrolases and two barley 1,3;1,4-β-D-glucan endohydrolases .
Active site labelling with epoxyalkyl β-D-oligoglucoside inhibitors identified Glu231 and Glu232 as the catalytic nucleophiles of the barley 1,3- and 1,3;1,4-β-D-glucan endohydrolases, respectively , located at the bottom of, and about two-thirds of the way along the substrate binding cleft. The general acid/base residue of 1,3;1,4-β-D-glucan endohydrolase was initially identified as Glu288 by chemical labelling procedures [3, 4], but this assignment was subsequently revised and Glu93 was proposed based on primary and tertiary structure similarity of GH17 enzymes with clan GH-A β-glycosidases [5, 6]. The 5-6 Å distance between Glu232 and Glu93 is typical of retaining enzymes.
The crystal structures of the barley 1,3-β-D-glucan endohydrolase isoenzyme GII and 1,3;1,4-β-D-glucan endohydrolase isoenzyme EII have been solved to 2.2-2.3 Å resolution and shown to adopt essentially identical (β/α)8 TIM barrel structures . The rms deviation in Cα positions between the two barley enzymes is 0.65 Å for 278 residues . This indicates that only minor differences in structure and amino acid dispositions at the substrate-binding and catalytic sites are sufficient to change a pre-existing 1,3-β-D-glucan endohydrolase into a highly specific 1,3;1,4-β-D-glucan endohydrolase .
A deep substrate-binding cleft extends across the surface of the enzyme and can accommodate 6-8 glucosyl-binding subsites . The open cleft enables the enzyme to bind at essentially any position along the 1,3;1,4-β-D-glucan substrate and hence to hydrolyse internal glycoside linkages. Like in other clan GH-A structures, the general acid/base and catalytic nucleophile glutamates are positioned on strands β-4 and β-7 [5, 6, 7].
The X-ray crystallographic data provide compelling evidence that the 1,3;1,4-β-D-glucan endohydrolases of barley evolved via the recruitment of pre-existing and widely distributed family GH17 1,3-β-D-glucan endohydrolases . The similarities in substrate specificity between family GH17 and GH16 enzymes has arisen through convergent evolution; the family GH16 enzymes are members of clan-B and have a β-jelly roll structure.
- First sterochemistry determination
- The stereochemical course of hydrolysis of Laminaria digitata laminarin and barley 1,3;1,4-β-glucan by barley 1,3-β-glucanase isoenzyme GII and 1,3;1,4-β-glucanase isoenzyme EII, respectively, was determined by 1H-NMR. Both enzymes catalyze hydrolysis with retention of anomeric configuration (e-->e) and may therefore operate via a double displacement mechanism .
- First catalytic nucleophile identification
- Active site labelling with epoxyalkyl-β-D-oligoglucoside inhibitors identified Glu231 and Glu232 as the catalytic nucleophiles of the barley 1,3- and 1,3;1,4-β-D-glucan endohydrolases, respectively .
- First general acid/base residue identification
- This residue was identified as the conserved Glu residue at the C-terminal end of strand β-4 based on primary and tertiary structure similarity of GH17 enzymes with clan GH-A β-glycosidases [5, 6].
- First 3-D structure
- Barley 1,3-β-D-glucan endohydrolase isoenzyme GII and 1,3;1,4-β-D-glucan endohydrolase isoenzyme EII, solved to 2.2-2.3 Å resolution .
Error fetching PMID 8514770:
Error fetching PMID 7713912:
Error fetching PMID 9649746:
Error fetching PMID 7624375:
Error fetching PMID 8146192:
Error fetching PMID 11554481:
- Woodward, J.R. and Fincher, G.B. (1982) Substrate specificities and kinetic properties of two (1-3),(1-4)-β-D-glucan endo-hydrolases from germinating barley (Hordeum vulgare). Carbohydr. Res. 106, 111-122. DOI:10.1016/S0008-6215(00)80737-5
- Error fetching PMID 7492591:
- Error fetching PMID 8514770:
- Error fetching PMID 7713912:
- Error fetching PMID 9649746:
- Error fetching PMID 7624375:
- Error fetching PMID 8146192:
- Error fetching PMID 11554481: