Glycoside Hydrolase Family 49

This page has been approved by the 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.

• Author: ^^^Takashi Tonozuka^^^
• Responsible Curator: ^^^Takashi Tonozuka^^^

---

Glycoside Hydrolase Family GHnn
Clan	GH-N
Mechanism	inverting
Active site residues	known
CAZy DB link
http://www.cazy.org/GH49.html

Substrate specificities

Glycoside hydrolases of family 49 cleave α-1,6-glucosidic linkages or α-1,4-glucosidic linkages of polysaccharides containing α-1,6-glucosidic linkages, dextran and pullulan. The major activities reported for this family of glycoside hydrolases are dextranase (EC 3.2.1.11), and a dextranase from Penicillium minioluteum, Dex49A, is currently the most characterised enzyme. Dextran 1,6-α-isomaltotriosidase (EC 3.2.1.95) [1] and isopullulanase (EC 3.2.1.57) activities have also been described.

Kinetics and Mechanism

Family GH49 α-glycosidases are inverting enzymes, as first shown by NMR on a dextranase Dex49A from Penicillium minioluteum [2] .

Catalytic Residues

Three Asp residues (Asp376, Asp395, and Asp396 in Dex49A) are conserved in the catalytic centre of members of Clan GH-N, GH49 and GH28 enzymes [2, 3], and all three of the Asp mutants of a GH49 enzyme, isopullulanase, lost their activities [4]. The general acid was first identified in Dex49A from Penicillium minioluteum as Asp395 following the three-dimensional structure determination. To date, it is unclear whether either (or both) of the Asp residues (Asp376 and Asp396 in Dex49A) acts as a general base in the reaction of GH49 and GH28 enzymes [2, 5, 6].

Three-dimensional structures

Two structures of GH49 enzymes are available so far [2, 3], and they display a two domain structure. The N-terminal domain is a β-sandwich and the C-terminal domain adopts a right-handed parallel β-helix. The similarity of the β-helix fold between GH49 and GH28 enzymes has been described, although almost none of the amino acid residues other than the three catalytic Asp residues is conserved between the two families [2, 3]. Each coil forming the cylindrical β-helix fold is composed of three β-sheets, which are named PB1, PB2, and PB3, following the original definition for a GH28 enzyme, pectate lyase C [7].

Family Firsts

First gene cloning

Dextranase from Arthrobacter sp. CB-8 [8].

First sterochemistry determination

Dextranase (Dex49A) from Penicillium minioluteum [2].

First general acid residue identification

Dextranase (Dex49A) from Penicillium minioluteum [2].

First 3-D structure

Dextranase (Dex49A) from Penicillium minioluteum by X-ray crystallography (PDB ID 1ogm) [2].

References

Error fetching PMID 10540747:
Error fetching PMID 12962629:
Error fetching PMID 18155243:
Error fetching PMID 15560783:
Error fetching PMID 8502994:
Error fetching PMID 1859672:

5. van Santen Y, Benen JA, Schröter KH, Kalk KH, Armand S, Visser J, and Dijkstra BW. (1999). 1.68-A crystal structure of endopolygalacturonase II from Aspergillus niger and identification of active site residues by site-directed mutagenesis. J Biol Chem. 1999;274(43):30474-80. DOI:10.1074/jbc.274.43.30474 |
6. Shimizu T, Nakatsu T, Miyairi K, Okuno T, and Kato H. (2002). Active-site architecture of endopolygalacturonase I from Stereum purpureum revealed by crystal structures in native and ligand-bound forms at atomic resolution. Biochemistry. 2002;41(21):6651-9. DOI:10.1021/bi025541a |