Characterization of β-1,2-Mannosyltransferase in Candida guilliermondii and Its Utilization in the Synthesis of Novel Oligosaccharides

Abstract

A particulate insoluble enzyme fraction containing mannosyltransferases from Candida guilliermondiiIFO 10279 strain cells was obtained as the residue after extracting a 105,000 × 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, Manβ1→2Manα1→3(Manα1→6)Manα1→2Manα1→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. A particulate insoluble enzyme fraction containing mannosyltransferases from Candida guilliermondiiIFO 10279 strain cells was obtained as the residue after extracting a 105,000 × 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, Manβ1→2Manα1→3(Manα1→6)Manα1→2Manα1→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. We have studied the structures of the cell wall mannans of medically important Candida species for several years (1Suzuki S. Shibata N. Kobayashi H. Latgé J.P. Boucias D. Fungal Cell Wall and Immune Response. Springer-Verlag, Heidelberg, Germany1991: 111-121Google Scholar, 2Hearn V.M. Cole G.T. Suzuki S. Van Regenmortel M.H.V. Structure of Antigens. CRC Press, Boca Raton, FL1993: 211-260Google Scholar, 3San-Blas G. Suzuki S. Hearn V. Pinel C. Kobayashi H. Mendez C. Nino G. Nishikawa A. San-Blas F. Shibata N. Shinoda T. Sorais F. Yamaguchi H. J. Med. Vet. Mycol. 1994; 32: 321-328Crossref PubMed Scopus (8) Google Scholar) and demonstrated that there are three types of β-1,2 linkage-containing side chains in its cell wall mannans. One is the β-1,2-linked mannooligomer, which is located in a phosphodiesterified oligosaccharide moiety, as the common epitope in the mannans of severalCandida species (4Shibata N. Ichikawa T. Tojo M. Takahashi M. Ito N. Okubo Y. Suzuki S. Arch. Biochem. Biophys. 1985; 243: 338-348Crossref PubMed Scopus (98) Google Scholar, 5Shibata N. Kobayashi H. Tojo M. Suzuki S. Arch. Biochem. Biophys. 1986; 251: 697-708Crossref PubMed Scopus (54) Google Scholar, 6Kobayashi H. Shibata N. Nakada M. Chaki S. Mizugami K. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1990; 278: 195-204Crossref PubMed Scopus (68) Google Scholar, 7Faille C. Michalski J.C. Strecker G. Mackenzie D.W.R. Camus D. Poulain D. Infect. Immun. 1990; 58: 2537-3544Crossref Google Scholar, 8Shibata N. Hisamichi K. Kikuchi T. Kobayashi H. Okawa Y. Suzuki S. Biochemistry. 1992; 31: 5680-5686Crossref PubMed Scopus (44) Google Scholar, 9Shibata N. Hisamichi K. Kobayashi H. Suzuki S. Arch. Biochem. Biophys. 1993; 302: 113-117Crossref PubMed Scopus (24) Google Scholar). The second type is β-1,2-linked mannose units attached to the nonreducing terminal of the α-1,2-linked oligomannosyl side chains in the mannans ofCandida albicans serotype A (10Shibata N. Fukasawa S. Kobayashi H. Tojo M. Yonezu T. Ambo A. Ohkubo Y. Suzuki S. Carbohydr. Res. 1989; 187: 239-253Crossref PubMed Scopus (77) Google Scholar, 11Kobayashi H. Shibata N. Mitobe H. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1989; 272: 364-375Crossref PubMed Scopus (84) Google Scholar), Candida tropicalis (12Kobayashi H. Matsuda K. Ikeda T. Suzuki M. Takahahi S. Suzuki A. Shibata N. Suzuki S. Infect. Immun. 1994; 62: 615-622Crossref PubMed Google Scholar), and Candida glabrata (13Kobayashi H. Mitobe M. Takahahi T. Yamamoto N. Shibata N. Suzuki S. Arch. Biochem. Biophys. 1992; 294: 662-669Crossref PubMed Scopus (39) Google Scholar). These two β-1,2 linkage-containing epitopes have been identified as corresponding to antigenic factors 5 (14Shibata N. Arai M. Haga E. Kikuchi T. Najima M. Satoh T. Kobayashi H. Suzuki S. Infect. Immun. 1992; 60: 4100-4110Crossref PubMed Google Scholar) and 6 (15Kobayashi H. Shibata N. Suzuki S. Infect. Immun. 1992; 60: 2106-2109Crossref PubMed Google Scholar), respectively. The third type of β-1,2 linkage-containing side chains can be observed in the mannan of Candida guilliermondii. This type of oligosaccharide contains β-1,2-linked mannose units attached to an α-1,3-linked mannose unit, the presence of which has been demonstrated in the cell wall mannans of Saccharomyces kluyveri (16Zhang W. Ballou C.E. J. Biol. Chem. 1981; 256: 10073-10079Abstract Full Text PDF PubMed Google Scholar, 17Shibata N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar), C. guilliermondii (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar), andCandida saitoana (19Shibata N. Onozawa M. Tadano N. Hinosawa Y. Suzuki A. Ikuta K. Kobayashi H. Suzuki S. Okawa Y. Arch. Biochem. Biophys. 1996; 336: 49-58Crossref PubMed Scopus (24) Google Scholar) and have been identified to correspond to antigenic factor 9 (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). The β-1,2 linkage-containing oligomannosyl side chains are the specific epitopes of the mannans of the genus Candida and have not been found in mammalian cells. Therefore, the detection of such an antigen in the sera from patients using immunological procedures is useful for the diagnosis of invasive candidiasis. Furthermore, several workers reported that the β-1,2 linkage-containing side chains participate in the adherence of fungal cells to host cells during the initial step of Candidainfection (20Miyakawa Y. Kuribayashi T. Kagaya K. Suzuki M. Nakase T. Fukasawa Y. Infect. Immum. 1992; 60: 2493-2499Crossref PubMed Google Scholar, 21Li R.-K. Cutler J.E. J. Biol. Chem. 1993; 268: 18293-18299Abstract Full Text PDF PubMed Google Scholar). Consequently, it is meaningful to characterize the β-1,2-mannosyltransferases responsible for the formation of the functional side chain. In a previous study (22Suzuki A. Takata Y. Oshie A. Tezuka A. Shibata N. Kobayashi H. Okawa Y. Suzuki S. FEBS Lett. 1995; 373: 275-279Crossref PubMed Scopus (26) Google Scholar), we detected the β-1,2-mannosyltransferase II, which is responsible for the transfer of an additional β-1,2-linked mannose unit to the nonreducing terminal β-1,2-linked one attached to the α-1,2-linked mannotetraose. Although we tried to detect the β-1,2-mannosyltransferase I responsible for the transfer of the first β-1,2-linked mannose unit to the α-1,2-linked mannotetraose to synthesize antigenic factor 6, we could not detect the activity in the homogenate of C. albicans serotype A cells. Therefore, we tried to detect another β-1,2-mannosyltransferase responsible for the formation of the antigenic factor 9, which have been found in the mannan of C. guilliermondii (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). A postulated biosynthetic pathway of the oligosaccharide corresponding to factor 9 can be depicted as shown in Fig. 1 based on the results obtained by the structural analysis of C. guilliermondii mannan (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar) and the substrate specificity of an α-1,6-mannosyltransferase of C. albicans (23Suzuki A. Shibata N. Suzuki M. Saitoh F. Takata Y. Oshie A. Oyamada H. Kobayashi H. Suzuki S. Okawa Y. Eur. J. Biochem. 1996; 240: 37-44Crossref PubMed Scopus Google Scholar) responsible for the formation of the oligosaccharides corresponding to factor N. Ikuta K. T. Satoh Y. Satoh R. Suzuki A. Kojima C. Kobayashi H. Hisamichi K. Suzuki S. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar, and Y. Eur. J. in Scholar). the of a β-1,2-linked mannose unit from GDP-mannose to the nonreducing terminal α-1,3-linked mannose unit of an side Manα1→3(Manα1→6)Manα1→2Manα1→2Man, synthesized from corresponding to factor and Y. FEBS Lett. Scholar), by of the β-1,2-mannosyltransferase and the of a β-1,2-linked one to a Manβ1→2Manα1→3(Manα1→6)Manα1→2Manα1→2Man, by of the β-1,2-mannosyltransferase in In the we tried to detect and characterize the β-1,2-mannosyltransferase using several oligosaccharide substrates prepared from various yeast mannans. The C. guilliermondii 10279 strain was the in a previous study (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). was obtained from and was from was from GDP-mannose and obtained from The × and obtained from The cell wall mannan of C. albicans was prepared by the by a (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). In this the oligosaccharides prepared from the mannans of C. S. and Candida with and respectively. Furthermore, the oligosaccharides synthesized by using albicans and C. guilliermondii cells by the of a and respectively. and Manα1→ prepared from C. albicans mannan N. Ikuta K. T. Satoh Y. Satoh R. Suzuki A. Kojima C. Kobayashi H. Hisamichi K. Suzuki S. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar), and from S. serotype mannan A. T. Ikeda R. Shinoda T. Y. 1990; PubMed Scopus Google Scholar), was from C. H. M. N. S. and Y. in and enzymatically prepared from corresponding and using the α-1,6-mannosyltransferase of C. albicans cells by the in a previous (23Suzuki A. Shibata N. Suzuki M. Saitoh F. Takata Y. Oshie A. Oyamada H. Kobayashi H. Suzuki S. Okawa Y. Eur. J. Biochem. 1996; 240: 37-44Crossref PubMed Scopus Google Scholar). The of the oligosaccharide was using the of Yamamoto S. S. S. T. J. Biochem. 1989; PubMed Scopus Google Scholar) as to an oligosaccharide of a prepared by 1 g of in of was the was at for The was and of the of a of of and of as the was The was and at for 1 The was with of and to a × of and from by with pH The of the fraction of C. guilliermondii strain cells was by the in a previous (22Suzuki A. Takata Y. Oshie A. Tezuka A. Shibata N. Kobayashi H. Okawa Y. Suzuki S. FEBS Lett. 1995; 373: 275-279Crossref PubMed Scopus (26) Google Scholar) as The cells in the yeast 1% and at the The cells and with 5 pH by The cells g of in of 5 pH containing and 1 and with a with g The homogenate was for at and the for at × the and for 1 at 105,000 × The pellet was in 1 of 5 pH containing 1% Triton and 1 and for at The was for at 105,000 × The pellet and at and for and The containing fraction of 5 GDP-mannose pH and Triton in a of was for 1 at The was by the of GDP-mannose and by the for at of the by was by as be The of was by its using as a was for the The was a of pH and and the was at of the was with and of and respectively. the NMR the enzyme was in a of containing of oligosaccharide mannan, fraction of pH and Triton X-100. incubation for at the was by and the was by The oligosaccharide was by and The mannan was and NMR using a at for in at a of was as the for the The was using the G.T. Biochem. 1985; PubMed Scopus Google Scholar) with as the The of in the and prepared from the C. guilliermondii 10279 strain cells and respectively. of the two by incubation with GDP-mannose and Although the two gave the corresponding to by the activity observed in fraction was three that in fraction Therefore, we fraction as the enzyme for the structure of the enzyme by a with was for The of the indicated that of the was transformed a The of the obtained by the enzyme as was by a sequential NMR using This has been demonstrated to results N. Hisamichi K. Kikuchi T. Kobayashi H. Okawa Y. Suzuki S. Biochemistry. 1992; 31: 5680-5686Crossref PubMed Scopus (44) Google Scholar, N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar, N. Ikuta K. T. Satoh Y. Satoh R. Suzuki A. Kojima C. Kobayashi H. Hisamichi K. Suzuki S. J. Biol. Chem. 1995; Full Text Full Text PDF PubMed Scopus Google Scholar, and Y. Eur. J. in Scholar, Ballou E. Ballou C.E. J. Biol. Chem. 1989; Full Text PDF PubMed Google Scholar). The of the terminal unit was to be Therefore, we the sequential of from the terminal mannose The in Fig. which by transfer and the with two mannose units. The on the the corresponding Fig. B and and of the the of the the of and the of was the the of which was from and the of was is by an the was found the an in to Therefore, we can the and the the of and the of we could the the of and the of of which the to the β-1,2-linked mannose unit (10Shibata N. Fukasawa S. Kobayashi H. Tojo M. Yonezu T. Ambo A. Ohkubo Y. Suzuki S. Carbohydr. Res. 1989; 187: 239-253Crossref PubMed Scopus (77) Google Scholar, 11Kobayashi H. Shibata N. Mitobe H. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1989; 272: 364-375Crossref PubMed Scopus (84) Google Scholar, 17Shibata N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar). this we could the from to as Therefore, we that of is by a β-1,2-linked mannose unit, has been shown that the of an mannose unit by a β-1,2-linked unit a (10Shibata N. Fukasawa S. Kobayashi H. Tojo M. Yonezu T. Ambo A. Ohkubo Y. Suzuki S. Carbohydr. Res. 1989; 187: 239-253Crossref PubMed Scopus (77) Google Scholar, 11Kobayashi H. Shibata N. Mitobe H. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1989; 272: 364-375Crossref PubMed Scopus (84) Google Scholar, 17Shibata N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar) and that the β-1,2-linked mannose units gave in a (14Shibata N. Arai M. Haga E. Kikuchi T. Najima M. Satoh T. Kobayashi H. Suzuki S. Infect. Immun. 1992; 60: 4100-4110Crossref PubMed Google Scholar, 17Shibata N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar, N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). In the of the of at at that of at and there is a specific with a These the that of is by the β-1,2-linked mannose Therefore, we the structure of to be the has been found in oligosaccharides (16Zhang W. Ballou C.E. J. Biol. Chem. 1981; 256: 10073-10079Abstract Full Text PDF PubMed Google Scholar) and N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar) of the mannan of S. that the is to detect the β-1,2-mannosyltransferase we of this The of pH on the activity was studied the pH of The β-1,2-mannosyltransferase activity at pH 6.0 in The of several and on the enzyme activity was studied using shown in the enzyme activity was by the of The enzyme activity was not by the of Furthermore, the enzyme activity was by the of and the activity could not be by the of of and on the β-1,2-mannosyltransferase activity in fraction incubation at with the of was the as that the additional ion to the was as incubation at with the of was in a The the as that the additional ion to the was as The enzyme activity is for at at the The of the β-1,2-mannosyltransferase for is shown in Fig. and the for from this and respectively. the substrate specificity of β-1,2-mannosyltransferase we the of the oligosaccharides prepared from the mannans of C. C. and S. and of enzymatically synthesized oligosaccharides as that the oligosaccharides containing an α-1,3-linked mannose unit at the nonreducing terminal can as Although and can as the of the β-1,2-mannosyltransferase and an from the structure of C. guilliermondii mannan, enzyme as and a at in the NMR not Therefore, it is that the mannose unit is attached to substrate with a β-1,2 (10Shibata N. Fukasawa S. Kobayashi H. Tojo M. Yonezu T. Ambo A. Ohkubo Y. Suzuki S. Carbohydr. Res. 1989; 187: 239-253Crossref PubMed Scopus (77) Google H. Shibata N. Mitobe H. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1989; 272: 364-375Crossref PubMed Scopus (84) Google Scholar, 17Shibata N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar). β-1,2 linkage-containing oligosaccharides have not been found in the of yeast it is to that we could novel oligosaccharides using the β-1,2-mannosyltransferase of C. guilliermondii. These results that the β-1,2-mannosyltransferase requires the nonreducing terminal α-1,3-linked mannose unit as the specificity of the β-1,2-mannosyltransferase and α-1,6-mannosyltransferase from the of the and enzyme 6 6 6 The the as that the substrate was as The β-1,2-mannosyltransferase and α-1,6-mannosyltransferase from the of the and enzyme respectively. in a The the as that the substrate was as Furthermore, to an by the of the β-1,2-mannosyltransferase we the enzyme using the β-1,2 linkage-containing as the and the structure of enzyme Fig. and the of The and of from to has been shown that the of an mannose unit by two β-1,2-linked units an (10Shibata N. Fukasawa S. Kobayashi H. Tojo M. Yonezu T. Ambo A. Ohkubo Y. Suzuki S. Carbohydr. Res. 1989; 187: 239-253Crossref PubMed Scopus (77) Google Scholar, 11Kobayashi H. Shibata N. Mitobe H. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1989; 272: 364-375Crossref PubMed Scopus (84) Google Scholar, 17Shibata N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar, N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). The of the of at at from that of at a at which to two β-1,2-linked mannose units (10Shibata N. Fukasawa S. Kobayashi H. Tojo M. Yonezu T. Ambo A. Ohkubo Y. Suzuki S. Carbohydr. Res. 1989; 187: 239-253Crossref PubMed Scopus (77) Google Scholar, 11Kobayashi H. Shibata N. Mitobe H. Ohkubo Y. Suzuki S. Arch. Biochem. Biophys. 1989; 272: 364-375Crossref PubMed Scopus (84) Google N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar, N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). The of the unit was by Therefore, we the structure of to be the results of and are shown in and of in this study residue 6 in a the of we a enzyme using the mannan of C. albicans serotype B which the nonreducing terminal α-1,3-linked mannose unit, not the β-1,2-linked mannose unit in the side the of the β-1,2-linked mannose unit to the mannan, we the structures of the and enzyme mannans using shown in Fig. 5 the gave and which correspond to the α-1,3-linked mannose units by one and two β-1,2-linked mannose N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar, N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar), with the of 9, and corresponding to the β-1,2-linked mannose units. the not 6 and corresponding to the α-1,2-linked mannose units by one and two β-1,2-linked mannose N. Kojima C. Satoh Y. Satoh R. Kobayashi H. Suzuki S. Eur. J. Biochem. 1993; 217: 1-12Crossref PubMed Scopus (30) Google Scholar, N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar). These results that the mannan was to structure by the of the the linkage-containing side chains in the mannan by the β-1,2-linked mannose units. of the on mannosyltransferases the of the cell wall of yeast have been W. Biophys. PubMed Scopus Google Scholar, T. Ballou C.E. S. A. PubMed Scopus Google Scholar, Ballou C.E. J. Biol. Chem. Full Text PDF PubMed Google Scholar, A. J. Biol. Chem. 1989; Full Text PDF PubMed Google Scholar, C.E. 1990; Scholar, Ballou C.E. J. Biol. Chem. Full Text PDF PubMed Google Scholar, K. T. Y. J. Y. J. 1992; PubMed Scopus Google Scholar, A. 1992; PubMed Scopus Google Scholar, A. Ballou Ballou C.E. S. A. 1992; PubMed Scopus Google Scholar, K. C. M. R. H. 1992; PubMed Google Scholar, Y. K. A. Y. Y. J. Biol. Chem. 1993; 268: Full Text PDF PubMed Google Scholar, F. T. S. A. 1994; PubMed Scopus Google Scholar, A. 1994; PubMed Scopus Google Scholar, M. A. H. J. Biol. Chem. 1996; 271: Full Text Full Text PDF PubMed Scopus Google Scholar, F. M. 1994; PubMed Scopus Google Scholar, 1995; PubMed Scopus Google Scholar), and of the structural for yeast mannosyltransferases have been the that the first mannose unit to a oligosaccharide K. T. Y. J. Y. J. 1992; PubMed Scopus Google Scholar, Y. K. A. Y. Y. J. Biol. Chem. 1993; 268: Full Text PDF PubMed Google is an A. 1992; PubMed Scopus Google Scholar), and is responsible for the of mannose units to and chains A. Ballou Ballou C.E. S. A. 1992; PubMed Scopus Google Scholar, K. C. M. R. H. 1992; PubMed Google Scholar). an F. T. S. A. 1994; PubMed Scopus Google Scholar), and is responsible for the of C.E. 1990; Scholar, F. T. S. A. 1994; PubMed Scopus Google Scholar). and found to corresponding to mannosyltransferases M. A. H. J. Biol. Chem. 1996; 271: Full Text Full Text PDF PubMed Scopus Google Scholar, F. M. 1994; PubMed Scopus Google Scholar). of for In a (19Shibata N. Onozawa M. Tadano N. Hinosawa Y. Suzuki A. Ikuta K. Kobayashi H. Suzuki S. Okawa Y. Arch. Biochem. Biophys. 1996; 336: 49-58Crossref PubMed Scopus (24) Google Scholar), we have reported the β-1,2-mannosyltransferase of C. the enzyme which one β-1,2-linked mannose unit to a β-1,2 linkage-containing side chain. there is study on the this is the first of the β-1,2-mannosyltransferase responsible for the of β-1,2-linked mannose unit to an The substrate specificity study that the β-1,2-mannosyltransferase requires the nonreducing terminal α-1,3-linked mannose the of the mannose unit not the substrate activity of the A substrate specificity has been found on the α-1,6-mannosyltransferase responsible for the of side chains (23Suzuki A. Shibata N. Suzuki M. Saitoh F. Takata Y. Oshie A. Oyamada H. Kobayashi H. Suzuki S. Okawa Y. Eur. J. Biochem. 1996; 240: 37-44Crossref PubMed Scopus Google Scholar). is of to the substrate of the two with Biophys. 1995; PubMed Scopus Google Scholar) with the α-1,3-linked K. A. M. Van A. Eur. J. Biochem. 1993; 217: PubMed Scopus (26) Google Scholar). C. guilliermondii cells the and from the structure of its mannan (18Shibata N. Akagi R. Hosoya T. Kawahara K. Suzuki A. Ikuta K. Kobayashi H. Hisamichi K. Okawa Y. Suzuki S. J. Biol. Chem. 1996; 271: 9259-9266Abstract Full Text Full Text PDF PubMed Scopus (45) Google Scholar), it is that we an linkage-containing oligosaccharide as the substrate of the enzyme we at two that the mannose Therefore, we the substrate specificity using of oligosaccharides with the mannose The substrate of oligosaccharides and for β-1,2-mannosyltransferase is in to that of the corresponding oligosaccharides and These results that the α-1,6-mannosyltransferase activity in fraction from C. guilliermondii is significantly the β-1,2-mannosyltransferase activity The for the of a β-1,2 linkage-containing side chains in the mannan of C. guilliermondii to be in this of the two the β-1,2 linkage-containing and 2Manα1→2Man, have been from the mannan of Candida saitoana (19Shibata N. Onozawa M. Tadano N. Hinosawa Y. Suzuki A. Ikuta K. Kobayashi H. Suzuki S. Okawa Y. Arch. Biochem. Biophys. 1996; 336: 49-58Crossref PubMed Scopus (24) Google Scholar), which has antigenic factor 9, it is that we an linkage-containing oligosaccharide with the enzyme prepared from this we β-1,2 linkage-containing In this we could synthesize a β-1,2 linkage-containing Manβ1→2Manα1→3(Manα1→6)Manα1→2Manα1→2Man, using fraction prepared from C. guilliermondii cells as the enzyme and as the substrate in the presence of and the of the oligosaccharides using specific not a and not it is an for the of We be to synthesize novel oligosaccharides by of the substrate specificity of enzyme obtained from several Candida The novel oligosaccharides to be useful for not the substrate specificity of mannosyltransferases the specificity of and Furthermore, this important for the of the chains of medically important The for by the enzyme was to reported for of S. W. Biophys. PubMed Scopus Google Scholar, T. Ballou C.E. S. A. PubMed Scopus Google Scholar, A. J. Biol. Chem. 1989; Full Text PDF PubMed Google Scholar). this of the enzyme is from that reported β-1,2-mannosyltransferase of C. which has for (22Suzuki A. Takata Y. Oshie A. Tezuka A. Shibata N. Kobayashi H. Okawa Y. Suzuki S. FEBS Lett. 1995; 373: 275-279Crossref PubMed Scopus (26) Google Scholar). In the study (22Suzuki A. Takata Y. Oshie A. Tezuka A. Shibata N. Kobayashi H. Okawa Y. Suzuki S. FEBS Lett. 1995; 373: 275-279Crossref PubMed Scopus (26) Google Scholar), we could not detect the β-1,2-mannosyltransferase I that is responsible for the of the first β-1,2-linked mannose unit to an α-1,2-linked mannotetraose side of C. Although the β-1,2-mannosyltransferase the nonreducing terminal α-1,3-linked mannose unit as identified in this the β-1,2-mannosyltransferase I not to a nonreducing terminal α-1,2-linked mannose This is the α-1,2-linked oligomannosyl of the β-1,2 linkage-containing side chains of C. albicans mannan is and that of the β-1,2 linkage-containing side chains of the mannans glabrata (13Kobayashi H. Mitobe M. Takahahi T. Yamamoto N. Shibata N. Suzuki S. Arch. Biochem. Biophys. 1992; 294: 662-669Crossref PubMed Scopus (39) Google Scholar) and Candida N. H. and Y. in are Therefore, the β-1,2-mannosyltransferases of species to the of the α-1,2-linked oligomannosyl side chains from the mannose to be for the of the of C. albicans and C. guilliermondii S. We K. the of of the of for the NMR