Acetolysis of the cell-wall mannan of Saccharomyces kluyveri under mild conditions, gave fragments with 1-6 mannose residues. The structures of mannopentaose and mannohexaose were determined to be Manal+3Manal-+2Manal+2Man 1 6
Manal and
Manfil+2Manal+3Mannl+2Manal+2Man, FManal respectively, by two-dimensional homonuclear Hartmann-Hahn spectroscopy and a sequential NMR assignment method that combines 'H-I3C correlated spectroscopy, relayed coherence transfer spectroscopy, 'H-detected heteronuclear multiple-bond connectivity and methylation analysis. The HI proton chemical shift of a neighboring a-1,2-linked mannose unit of the 3-0-substituted structure was shifted upfield by the addition of a mannose unit to the adjacent 3-0-substituted unit by an a-1,6 linkage. The characteristic H1 -H2-correlated cross-peak of the a-1,3-linked mannose unit substituted by a B1,2 linkage, pl+2Mana1+3, in the mannan of S. kluyveri, as also found by twodimensional homonuclear Hartmann-Hahn spectroscopy in the rnannan of Cundidu guilliermondii, a pathogenic yeast in man.In a previous study, it was found that mannans of the pathogenic yeast, Candida albicans, strains NIH A-207 (serotype A), NIH B-792 (serotype B) and J-1012 (serotype A) all contained /3-1,2-linked mannose units [l -81. Furthermore, we defined that a P-1,2-linked mannose unit is linked to an a-1,2-linked unit, Manfil+2Manal+2Manal+ andManpl-+2Manpl+2Manal-+2Manal-+, and that these specific structures are present in serotype-A epitopes corresponding to antigenic factor 6 in C. albicans species [9]. The phosphodiesterified p-1,2-linked oligomannosyl residues, Man,!ll+(2Manpl+),2Man (n = 0-5), are common feaCorrespondence to S. Suzuh, The Second Department of Hygienic Chemistry, Tohoku College of Pharmacy, 4-4-1 Komatsushima, Sendai Aoba-ku, Miyagi, Japan 981 Abbreviations. 2D, two-dimensional; HOHAHA, homonuclear Hartmann-Hahn spectroscopy ; COSY, correlated spectroscopy ; relayed COSY, relayed coherence transfer spectroscopy; HMBC, 'Hdetected heteronuclear multiple-bond connectivity; Fr. S, alkalimodified mannan: SM,, mannooligosaccharide obtained after mild acetolysis, where x indicates number of mannose residues; SM,-e, a-rnannosidase-resistant hexaose; SM,-eH, NaBH,-reduction product of SM,-e. tures present in the mannans of strains of both serotypes A and €3, and correspond to antigenic factor 5 [lo].However, Zhang and Ballou [111 reported the presence of a fi-1,2-linked mannose unit, in the oligomannosyl moiety which can be /? eliminated, for the 0-glycosylated form. This unit is linked to serine and/or threonine in the peptide moiety of a mannoprotein of Saccharomyces kluyveri. In this study [l I], the structure of the oligomannosyl residue containing the p-1,2 linkage was determined to be Manpl+2Mana 1 +3Manal+2Manal+2Manal +Ser/Thr
I6Manal .