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The structural basis of Erwinia rhapontici isomaltulose synthase.

<div class='caption-title'>Schematic depictions of the sucrose isomerization process.</div>
<div class='caption-title'>The crystal structure of native NX-5.</div>
<div class='caption-title'>A stereo view of the binding mode between sucrose and NX-5 protein in the E295Q/sucrose complex.</div>
Xu Z, Li S, Li J, Li Y, Feng X, Wang R, Xu H, Zhou J
OpenAccess logo PLoS One 8 e74788 (2013)
Related entries: 4how, 4hph

Cited: 12 times
EuropePMC logo PMID: 24069347

Abstract

Sucrose isomerase NX-5 from Erwiniarhapontici efficiently catalyzes the isomerization of sucrose to isomaltulose (main product) and trehalulose (by-product). To investigate the molecular mechanism controlling sucrose isomer formation, we determined the crystal structures of native NX-5 and its mutant complexes E295Q/sucrose and D241A/glucose at 1.70 Å, 1.70 Å and 2.00 Å, respectively. The overall structure and active site architecture of NX-5 resemble those of other reported sucrose isomerases. Strikingly, the substrate binding mode of NX-5 is also similar to that of trehalulose synthase from Pseudomonasmesoacidophila MX-45 (MutB). Detailed structural analysis revealed the catalytic RXDRX motif and the adjacent 10-residue loop of NX-5 and isomaltulose synthase PalI from Klebsiella sp. LX3 adopt a distinct orientation from those of trehalulose synthases. Mutations of the loop region of NX-5 resulted in significant changes of the product ratio between isomaltulose and trehalulose. The molecular dynamics simulation data supported the product specificity of NX-5 towards isomaltulose and the role of the loop(330-339) in NX-5 catalysis. This work should prove useful for the engineering of sucrose isomerase for industrial carbohydrate biotransformations.

Articles - 4hoz mentioned but not cited (1)

  1. The structural basis of Erwinia rhapontici isomaltulose synthase. Xu Z, Li S, Li J, Li Y, Feng X, Wang R, Xu H, Zhou J. PLoS One 8 e74788 (2013)


Reviews citing this publication (2)

  1. Biotechnical production of trehalose through the trehalose synthase pathway: current status and future prospects. Cai X, Seitl I, Mu W, Zhang T, Stressler T, Fischer L, Jiang B. Appl Microbiol Biotechnol 102 2965-2976 (2018)
  2. Analyzing Current Trends and Possible Strategies to Improve Sucrose Isomerases' Thermostability. Sardiña-Peña AJ, Mesa-Ramos L, Iglesias-Figueroa BF, Ballinas-Casarrubias L, Siqueiros-Cendón TS, Espinoza-Sánchez EA, Flores-Holguín NR, Arévalo-Gallegos S, Rascón-Cruz Q. Int J Mol Sci 24 14513 (2023)

Articles citing this publication (9)

  1. Insight into the substrate specificity change caused by the Y227H mutation of α-glucosidase III from the European honeybee (Apis mellifera) through molecular dynamics simulations. Na Ayutthaya PP, Chanchao C, Chunsrivirot S. PLoS One 13 e0198484 (2018)
  2. Molecular dynamics reveals insight into how N226P and H227Y mutations affect maltose binding in the active site of α-glucosidase II from European honeybee, Apis mellifera. Punnatin P, Chanchao C, Chunsrivirot S. PLoS One 15 e0229734 (2020)
  3. Enhancing the Thermostability of Serratia plymuthica Sucrose Isomerase Using B-Factor-Directed Mutagenesis. Duan X, Cheng S, Ai Y, Wu J. PLoS One 11 e0149208 (2016)
  4. Structure-function analysis of silkworm sucrose hydrolase uncovers the mechanism of substrate specificity in GH13 subfamily 17 exo-α-glucosidases. Miyazaki T, Park EY. J Biol Chem 295 8784-8797 (2020)
  5. Bacillus licheniformis trehalose-6-phosphate hydrolase structures suggest keys to substrate specificity. Lin MG, Chi MC, Naveen V, Li YC, Lin LL, Hsiao CD. Acta Crystallogr D Struct Biol 72 59-70 (2016)
  6. Engineering a Highly Active Sucrose Isomerase for Enhanced Product Specificity by Using a "Battleship" Strategy. Pilak P, Schiefner A, Seiboth J, Oehrlein J, Skerra A. Chembiochem 21 2161-2169 (2020)
  7. Characterization of divergent pseudo-sucrose isomerase from Azotobacter vinelandii: Deciphering the absence of sucrose isomerase activity. Jung JH, Kim MJ, Jeong WS, Seo DH, Ha SJ, Kim YW, Park CS. Biochem Biophys Res Commun 483 115-121 (2017)
  8. Sustainable isomaltulose production in Corynebacterium glutamicum by engineering the thermostability of sucrose isomerase coupled with one-step simplified cell immobilization. Hu M, Liu F, Wang Z, Shao M, Xu M, Yang T, Zhang R, Zhang X, Rao Z. Front Microbiol 13 979079 (2022)
  9. Thermostability improvement of sucrose isomerase PalI NX-5: a comprehensive strategy. Sardiña-Peña AJ, Ballinas-Casarrubias L, Siqueiros-Cendón TS, Espinoza-Sánchez EA, Flores-Holguín NR, Iglesias-Figueroa BF, Rascón-Cruz Q. Biotechnol Lett 45 885-904 (2023)


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