Isomaltulose synthase (PalI) of Klebsiella sp. LX3: Crystal structure and implication of mechanism
Zhang, D. Li, N. Lok, S.-M. Swaminathan, K. Zhang, L.-H.
Li, N.
Lok, S.-M.
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Abstract
Isomaltulose synthase from Klebsiella sp. LX3 (PalI, EC 5.4.99.11) catalyzes the isomerization of sucrose to produce isomaltulose (α-D-glucosylpyranosyl-1,6-D-fructofuranose) and trehalulose (α-D-glucosylpyranosyl-1,1-D-fructofuranose). The PalI structure, solved at 2.2-Å resolution with an R-factor of 19.4% and Rfree of 24.2%, consists of three domains: an N-terminal catalytic (β/α) 8 domain, a subdomain between Nβ3 and Nα3, and a C-terminal domain having seven β-strands. The active site architecture of PalI is identical to that of other glycoside hydrolase family 13 members, suggesting a similar mechanism in substrate binding and hydrolysis. However, a unique RLDRD motif in the proximity of the active site has been identified and shown biochemically to be responsible for sucrose isomerization. A two-step reaction mechanism for hydrolysis and isomerization, which occurs in the same pocket is proposed based on both the structural and biochemical data. Selected C-terminal truncations have been shown to reduce and even abolish the enzyme activity, consistent with the predicted role of the C-terminal residues in the maintenance of enzyme conformation and active site topology.
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Journal of Biological Chemistry
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Date
2003-09-12
DOI
10.1074/jbc.M302616200
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Article