1y4w | pdb_00001y4w
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Crystal structure of exo-inulinase from Aspergillus awamori in spacegroup P21
Overview
Exo-inulinases hydrolyze terminal, non-reducing 2,1-linked and 2,6-linked beta-d-fructofuranose residues in inulin, levan and sucrose releasing beta-d-fructose. We present the X-ray structure at 1.55A resolution of exo-inulinase from Aspergillus awamori, a member of glycoside hydrolase family 32, solved by single isomorphous replacement with the anomalous scattering method using the heavy-atom sites derived from a quick cryo-soaking technique. The tertiary structure of this enzyme folds into two domains: the N-terminal catalytic domain of an unusual five-bladed beta-propeller fold and the C-terminal domain folded into a beta-sandwich-like structure. Its structural architecture is very similar to that of another member of glycoside hydrolase family 32, invertase (beta-fructosidase) from Thermotoga maritima, determined recently by X-ray crystallography The exo-inulinase is a glycoprotein containing five N-linked oligosaccharides. Two crystal forms obtained under similar crystallization conditions differ by the degree of protein glycosylation. The X-ray structure of the enzyme:fructose complex, at a resolution of 1.87A, reveals two catalytically important residues: Asp41 and Glu241, a nucleophile and a catalytic acid/base, respectively. The distance between the side-chains of these residues is consistent with a double displacement mechanism of reaction. Asp189, which is part of the Arg-Asp-Pro motif, provides hydrogen bonds important for substrate recognition.
About this Structure
1Y4W is a Single protein structure of sequence from Aspergillus awamori with NAG and GOL as ligands. Active as Fructan beta-fructosidase, with EC number 3.2.1.80 Full crystallographic information is available from OCA.
Reference
Crystal structure of exo-inulinase from Aspergillus awamori: the enzyme fold and structural determinants of substrate recognition., Nagem RA, Rojas AL, Golubev AM, Korneeva OS, Eneyskaya EV, Kulminskaya AA, Neustroev KN, Polikarpov I, J Mol Biol. 2004 Nov 19;344(2):471-80. PMID:15522299
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- Aspergillus awamori
- Fructan beta-fructosidase
- Single protein
- Eneyskaya, E V.
- Golubev, A M.
- Korneeva, O S.
- Kulminskaya, A A.
- Nagem, R A.P.
- Neustroev, K N.
- Polikarpov, I.
- Rojas, A L.
- GOL
- NAG
- Crystallographic structure
- Exo-inulinase
- Glycoside hydrolase family 32
- Native structure
- X-ray structure
