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Enrico Cabib - One of the best experts on this subject based on the ideXlab platform.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1997Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170–1178), the linkage region between chitin and β(1→3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with β(1→3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, β(1→3)-glucan, β(1→6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to β(1→6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five α-linked mannosyl residues. The β(1→6)-glucan has some β(1→3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a β(1→4) or β(1→2) linkage. Finally, the reducing end of β(1→6)-glucan is connected to the nonreducing terminal glucose of β(1→3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and β(1→6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1995Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170-1178), the linkage region between chitin and beta(1-->3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with beta(1-->3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, beta(1-->3)-glucan, beta(1-->6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to beta(1-->6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five alpha-linked mannosyl residues. The beta(1-->6)-glucan has some beta(1-->3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a beta(1-->4) or beta(1-->2) linkage. Finally, the reducing end of beta(1-->6)-glucan is connected to the nonreducing terminal glucose of beta(1-->3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and beta(1-->6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
Roman Kollar - One of the best experts on this subject based on the ideXlab platform.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1997Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170–1178), the linkage region between chitin and β(1→3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with β(1→3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, β(1→3)-glucan, β(1→6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to β(1→6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five α-linked mannosyl residues. The β(1→6)-glucan has some β(1→3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a β(1→4) or β(1→2) linkage. Finally, the reducing end of β(1→6)-glucan is connected to the nonreducing terminal glucose of β(1→3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and β(1→6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1995Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170-1178), the linkage region between chitin and beta(1-->3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with beta(1-->3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, beta(1-->3)-glucan, beta(1-->6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to beta(1-->6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five alpha-linked mannosyl residues. The beta(1-->6)-glucan has some beta(1-->3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a beta(1-->4) or beta(1-->2) linkage. Finally, the reducing end of beta(1-->6)-glucan is connected to the nonreducing terminal glucose of beta(1-->3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and beta(1-->6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
Akihiko Kondo - One of the best experts on this subject based on the ideXlab platform.
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novel strategy for anchorage position control of gpi attached proteins in the Yeast Cell wall using different gpi anchoring domains
Metabolic Engineering, 2020Co-Authors: Kentaro Inokuma, Tomohisa Hasunuma, Hiroki Kurono, Riaan Den Haan, Willem H Van Zyl, Akihiko KondoAbstract:The Yeast Cell surface provides space to display functional proteins. Heterologous proteins can be covalently anchored to the Yeast Cell wall by fusing them with the anchoring domain of glycosylphosphatidylinositol (GPI)-anchored Cell wall proteins (GPI-CWPs). In the Yeast Cell-surface display system, the anchorage position of the target protein in the Cell wall is an important factor that maximizes the capabilities of engineered Yeast Cells because the Yeast Cell wall consists of a 100- to 200-nm-thick microfibrillar array of glucan chains. However, knowledge is limited regarding the anchorage position of GPI-attached proteins in the Yeast Cell wall. Here, we report a comparative study on the effect of GPI-anchoring domain-heterologous protein fusions on Yeast Cell wall localization. GPI-anchoring domains derived from well-characterized GPI-CWPs, namely Sed1p and Sag1p, were used for the Cell-surface display of heterologous proteins in the Yeast Saccharomyces cerevisiae. Immunoelectron-microscopic analysis of enhanced green fluorescent protein (eGFP)-displaying Cells revealed that the anchorage position of the GPI-attached protein in the Cell wall could be controlled by changing the fused anchoring domain. eGFP fused with the Sed1-anchoring domain predominantly localized to the external surface of the Cell wall, whereas the anchorage position of eGFP fused with the Sag1-anchoring domain was mainly inside the Cell wall. We also demonstrate the application of the anchorage position control technique to improve the Cellulolytic ability of Cellulase-displaying Yeast. The ethanol titer during the simultaneous saccharification and fermentation of hydrothermally-processed rice straw was improved by 30% after repositioning the exo- and endo-Cellulases using Sed1- and Sag1-anchor domains. This novel anchorage position control strategy will enable the efficient utilization of the Cell wall space in various fields of Yeast Cell-surface display technology.
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mapping of endoglucanases displayed on Yeast Cell surface using atomic force microscopy
Colloids and Surfaces B: Biointerfaces, 2017Co-Authors: Musashi Takenaka, Chiaki Ogino, Tomohisa Hasunuma, Takuya Kobayashi, Kentaro Inokuma, Tatsuo Maruyama, Akihiko KondoAbstract:The surface of Yeast Cells has been an attractive interface for the effective use of Cellulose. Surface enzymes, however, are difficult to visualize and evaluate. In this study, two kinds of unique anchoring regions were used to display the Cellulase, endoglucanase (EG), on a Yeast Cell surface. Differences in the display level and the localization of EG were observed by atomic force microscopy. By surveying the Yeast Cell surface with a chemically modified cantilever, the interactive force between the Cellulose and EG was measured. Force curve mapping revealed differences in the display levels and the localization of EG according to anchoring regions. The proposed methodology enables visualization of displayed enzymes such as EG on the Yeast Cell surface.
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mapping of endoglucanases displayed on Yeast Cell surface using atomic force microscopy b biointerfaces
Colloids and Surfaces, 2017Co-Authors: Musashi Takenaka, Chiaki Ogino, Tomohisa Hasunuma, Takuya Kobayashi, Kentaro Inokuma, Tatsuo Maruyama, Akihiko KondoAbstract:The surface of Yeast Cells has been an attractive interface for the effective use of Cellulose. Surface enzymes, however, are difficult to visualize and evaluate. In this study, two kinds of unique anchoring regions were used to display the Cellulase, endoglucanase (EG), on a Yeast Cell surface. Differences in the display level and the localization of EG were observed by atomic force microscopy. By surveying the Yeast Cell surface with a chemically modified cantilever, the interactive force between the Cellulose and EG was measured. Force curve mapping revealed differences in the display levels and the localization of EG according to anchoring regions. The proposed methodology enables visualization of displayed enzymes such as EG on the Yeast Cell surface.
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synergetic effect of Yeast Cell surface expression of Cellulase and expansin like protein on direct ethanol production from Cellulose
Microbial Cell Factories, 2013Co-Authors: Yuki Nakatani, Ryosuke Yamada, Chiaki Ogino, Akihiko KondoAbstract:Numerous studies have examined the direct fermentation of Cellulosic materials by Cellulase-expressing Yeast; however, ethanol productivity in these systems has not yet reached an industrial level. Certain microorganisms, such as the Cellulolytic fungus Trichoderma reesei, produce expansin-like proteins, which have a Cellulose-loosening effect that may increase the breakdown of Cellulose. Here, to improve the direct conversion of Cellulose to ethanol, Yeast Saccharomyces cerevisiae co-displaying Cellulase and expansin-like protein on the Cell surface were constructed and examined for direct ethanol fermentation performance. The Cellulase and expansin-like protein co-expressing strain showed 246 mU/g-wet Cell of phosphoric acid swollen Cellulose (PASC) degradation activity, which corresponded to 2.9-fold higher activity than that of a Cellulase-expressing strain. This result clearly demonstrated that Yeast Cell-surface expressed Cellulase and expansin-like protein act synergistically to breakdown Cellulose. In fermentation experiments examining direct ethanol production from PASC, the Cellulase and expansin-like protein co-expressing strain produced 3.4 g/L ethanol after 96 h of fermentation, a concentration that was 1.4-fold higher than that achieved by the Cellulase-expressing strain (2.5 g/L). The PASC degradation and fermentation ability of an engineered Yeast strain was markedly improved by co-expressing Cellulase and expansin-like protein on the Cell surface. To our knowledge, this is the first report to demonstrate the synergetic effect of co-expressing Cellulase and expansin-like protein on a Yeast Cell surface, which may be a promising strategy for constructing direct ethanol fermenting Yeast from Cellulose.
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display of a functional hetero oligomeric catalytic antibody on the Yeast Cell surface
Applied Microbiology and Biotechnology, 2003Co-Authors: Ying Lin, Seizaburo Shiraga, Takeshi Tsumuraya, Akihiko Kondo, T Wakabayashi, I Fujii, Mitsuyoshi UedaAbstract:A functional hetero-oligomeric protein was, for the first time, displayed on the Yeast Cell surface. A hetero-oligomeric Fab fragment of the catalytic antibody 6D9 can hydrolyze a non-bioactive chloramphenicol monoester derivative to produce chloramphenicol. The gene encoding the light chain of the Fab fragment of 6D9 was expressed with the tandemly-linked C-terminal half of α-agglutinin. At the same time, the gene encoding the Fd fragment of the heavy chain of the Fab fragment was expressed as a secretion protein. The combined Fab fragment displayed and associated on the Yeast Cell surface had an intermolecular disulfide linkage between the light and heavy chains. This protein fragment catalyzed the hydrolysis of a chloramphenicol monoester derivative and exhibited high stability in binding with a transition-state analog (TSA). The catalytic reaction was also inhibited by the TSA. The successful display of a functional hetero-oligomeric catalytic antibody provides a useful model for the display of hetero-oligomeric proteins and enzymes.
J C Kapteyn - One of the best experts on this subject based on the ideXlab platform.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1997Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170–1178), the linkage region between chitin and β(1→3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with β(1→3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, β(1→3)-glucan, β(1→6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to β(1→6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five α-linked mannosyl residues. The β(1→6)-glucan has some β(1→3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a β(1→4) or β(1→2) linkage. Finally, the reducing end of β(1→6)-glucan is connected to the nonreducing terminal glucose of β(1→3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and β(1→6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1995Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170-1178), the linkage region between chitin and beta(1-->3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with beta(1-->3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, beta(1-->3)-glucan, beta(1-->6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to beta(1-->6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five alpha-linked mannosyl residues. The beta(1-->6)-glucan has some beta(1-->3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a beta(1-->4) or beta(1-->2) linkage. Finally, the reducing end of beta(1-->6)-glucan is connected to the nonreducing terminal glucose of beta(1-->3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and beta(1-->6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
Jana Drgonova - One of the best experts on this subject based on the ideXlab platform.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1997Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170–1178), the linkage region between chitin and β(1→3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with β(1→3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, β(1→3)-glucan, β(1→6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to β(1→6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five α-linked mannosyl residues. The β(1→6)-glucan has some β(1→3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a β(1→4) or β(1→2) linkage. Finally, the reducing end of β(1→6)-glucan is connected to the nonreducing terminal glucose of β(1→3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and β(1→6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
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architecture of the Yeast Cell wall beta 1 6 glucan interconnects mannoprotein beta 1 3 glucan and chitin
Journal of Biological Chemistry, 1995Co-Authors: Roman Kollar, Frans M Klis, Bruce B Reinhold, Eva Petrakova, Herman J C Yeh, Gilbert Ashwell, Jana Drgonova, J C Kapteyn, Enrico CabibAbstract:In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170-1178), the linkage region between chitin and beta(1-->3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of Yeast Cell walls with beta(1-->3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the Cell wall, beta(1-->3)-glucan, beta(1-->6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to beta(1-->6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five alpha-linked mannosyl residues. The beta(1-->6)-glucan has some beta(1-->3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a beta(1-->4) or beta(1-->2) linkage. Finally, the reducing end of beta(1-->6)-glucan is connected to the nonreducing terminal glucose of beta(1-->3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and beta(1-->6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the Yeast Cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed.
