Practical preparation of UDP-apiose and its applications for studying apiosyltransferase

Carbohydr Res. 2019 May 15:477:20-25. doi: 10.1016/j.carres.2019.03.011. Epub 2019 Mar 23.

Abstract

UDP-apiose, a donor substrate of apiosyltransferases, is labile because of its intramolecular self-cyclization ability, resulting in the formation of apiofuranosyl-1,2-cyclic phosphate. Therefore, stabilization of UDP-apiose is indispensable for its availability and identifying and characterizing the apiosyltransferases involved in the biosynthesis of apiosylated sugar chains and glycosides. Here, we established a method for stabilizing UDP-apiose using bulky cations as counter ions. Bulky cations such as triethylamine effectively suppressed the degradation of UDP-apiose in solution. The half-life of UDP-apiose was increased to 48.1 ± 2.4 h at pH 6.0 and 25 °C using triethylamine as a counter cation. UDP-apiose coordinated with a counter cation enabled long-term storage under freezing conditions. UDP-apiose was utilized as a donor substrate for apigenin 7-O-β-D-glucoside apiosyltransferase to produce the apiosylated glycoside apiin. This apiosyltransferase assay will be useful for identifying genes encoding apiosyltransferases.

Keywords: Apiin; Apiose; Glycoside; Glycosyltransferase; Nucleotide sugar; UDP-apiose.

MeSH terms

  • Carbohydrate Conformation
  • Enzyme Assays / methods*
  • Pentosyltransferases / genetics
  • Pentosyltransferases / metabolism*
  • Uridine Diphosphate Sugars / chemical synthesis*
  • Uridine Diphosphate Sugars / chemistry
  • Uridine Diphosphate Sugars / metabolism*

Substances

  • UDP-apiose
  • Uridine Diphosphate Sugars
  • Pentosyltransferases