TY - JOUR
T1 - Characterization of β-1,2-mannosyltransferase in Candida guilliermondii and its utilization in the synthesis of novel oligosaccharides
AU - Suzuki, Akifumi
AU - Shibata, Nobuyuki
AU - Suzuki, Mikiko
AU - Saitoh, Fumie
AU - Oyamada, Hiroko
AU - Kobayashi, Hidemitsu
AU - Suzuki, Shigeo
AU - Okawa, Yoshio
PY - 1997/7/4
Y1 - 1997/7/4
N2 - 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.
AB - 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.
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U2 - 10.1074/jbc.272.27.16822
DO - 10.1074/jbc.272.27.16822
M3 - Article
C2 - 9201988
AN - SCOPUS:0030811995
VL - 272
SP - 16822
EP - 16828
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 27
ER -