Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis.

D. Sprogoe, L.A.M. van den Broek, O. Mirza, J.S. Kastrup, A.G.J. Voragen, M. Gajhede, L.K. Skov

Research output: Contribution to journalArticleAcademicpeer-review

67 Citations (Scopus)

Abstract

Around 80 enzymes are implicated in the generic starch and sucrose pathways. One of these enzymes is sucrose phosphorylase, which reversibly catalyzes the conversion of sucrose and orthophosphate to d-Fructose and a-d-glucose 1-phosphate. Here, we present the crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis (BiSP) refined at 1.77 Å resolution. It represents the first 3D structure of a sucrose phosphorylase and is the first structure of a phosphate-dependent enzyme from the glycoside hydrolase family 13. The structure of BiSP is composed of the four domains A, B, B‘, and C. Domain A comprises the (ß/a)8-barrel common to family 13. The catalytic active-site residues (Asp192 and Glu232) are located at the tips of ß-sheets 4 and 5 in the (ß/a)8-barrel, as required for family 13 members. The topology of the B‘ domain disfavors oligosaccharide binding and reduces the size of the substrate access channel compared to other family 13 members, underlining the role of this domain in modulating the function of these enzymes. It is remarkable that the fold of the C domain is not observed in any other known hydrolases of family 13. BiSP was found as a homodimer in the crystal, and a dimer contact surface area of 960 Å2 per monomer was calculated. The majority of the interactions are confined to the two B domains, but interactions between the loop 8 regions of the two barrels are also observed. This results in a large cavity in the dimer, including the entrance to the two active sites.
Original languageEnglish
Pages (from-to)1156-1162
JournalBiochemistry
Volume43
Issue number5
DOIs
Publication statusPublished - 2004

Keywords

  • neisseria-polysaccharea
  • glycosyl hydrolases
  • sequence alignment
  • maltogenic amylase
  • alpha-amylase
  • amylosucrase
  • crystallization
  • crystallography
  • neopullulanase
  • purification

Fingerprint Dive into the research topics of 'Crystal structure of sucrose phosphorylase from Bifidobacterium adolescentis.'. Together they form a unique fingerprint.

Cite this