Characterization of β-1,2-mannosyltransferase in Candida guilliermondii and its utilization in the synthesis of novel oligosaccharides

Akifumi Suzuki, Nobuyuki Shibata, Mikiko Suzuki, Fumie Saitoh, Hiroko Oyamada, Hidemitsu Kobayashi, Shigeo Suzuki, Yoshio Okawa

Research output: Contribution to journalArticlepeer-review

16 Citations (Scopus)

Abstract

A particulate insoluble enzyme fraction containing mannosyltransferases from Candida guilliermondii IFO 10279 strain cells was obtained as the residue after extracting a 105,000 x g pellet of cell homogenate with 1% Triton X-100. Incubation of this fraction with a mannopentaose, Manα1→3(Manα1→6)Manα1→2Manα1→2Man, in the presence of GDP-mannose and Mn2+ ion at pH 6.0 gave a third type of β-1,2 linkage-containing mannohexaose, Man β1→2Manα1→3(Manα1→6)Manα1→2Man-α1→2Man, the structure of which was identified by means of a sequential NMR assignment. The results of a substrate specificity study indicated that the β-1,2- mannosyltransferase requires a mannobiosyl unit, Manα1→3Manα1→, at the nonreducing terminal site. We synthesized novel oligosaccharides using substrates possessing a nonreducing terminal α-1,3-linked mannose unit prepared from various yeast mannans. Further incubation of the enzymatically synthesized oligosaccharide with the enzyme fraction gave the following structure, Manβ1→2Manβ1→2Manα1→3(Manα1→6)Manα1→2Manα1→2Man, which has been found to correspond to antigenic factor 9. Incubation of Candida albicans serotype B mannan with the enzyme fraction gave significantly transformed mannan, which contains the third type of β-1,2-linked mannose units.

Original languageEnglish
Pages (from-to)16822-16828
Number of pages7
JournalJournal of Biological Chemistry
Volume272
Issue number27
DOIs
Publication statusPublished - 1997 Jul 4
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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