caa a22 4500 445325879 CHVBK 20180317142720.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s10858-011-9525-1 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9525-1 Overexpression of a homogeneous oligosaccharide with 13C labeling by genetically engineered yeast strain [Elektronische Daten] [Yukiko Kamiya, Sayoko Yamamoto, Yasunori Chiba, Yoshifumi Jigami, Koichi Kato] This report describes a novel method for overexpression of 13C-labeled oligosaccharides using genetically engineered Saccharomyces cerevisiae cells, in which a homogeneous high-mannose-type oligosaccharide accumulates because of deletions of genes encoding three enzymes involved in the processing pathway of asparagine-linked oligosaccharides in the Golgi complex. Using uniformly 13C-labeled glucose as the sole carbon source in the culture medium of these engineered yeast cells, high yields of the isotopically labeled Man8GlcNAc2 oligosaccharide could be successfully harvested from glycoprotein extracts of the cells. Furthermore, 13C labeling at selected positions of the sugar residues in the oligosaccharide could be achieved using a site-specific 13C-enriched glucose as the metabolic precursor, facilitating NMR spectral assignments. The 13C-labeling method presented provides the technical basis for NMR analyses of structures, dynamics, and interactions of larger, branched oligosaccharides. Springer Science+Business Media B.V., 2011 Oligosaccharide nationallicence Stable isotope labeling nationallicence Engineered yeast nationallicence Saccharomyces cerevisiae nationallicence NMR structural glycobiology nationallicence Kamiya Yukiko Okazaki Institute for Integrative Bioscience and Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, 444-8787, Okazaki, Aichi, Japan aut Yamamoto Sayoko Okazaki Institute for Integrative Bioscience and Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, 444-8787, Okazaki, Aichi, Japan aut Chiba Yasunori Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Tukuba Central 6, 1-1-1 Higashi, 305-8566, Tukuba, Ibaraki, Japan aut Jigami Yoshifumi Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Tukuba Central 6, 1-1-1 Higashi, 305-8566, Tukuba, Ibaraki, Japan aut Kato Koichi Okazaki Institute for Integrative Bioscience and Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, 444-8787, Okazaki, Aichi, Japan aut Journal of Biomolecular NMR Springer Netherlands 50/4(2011-08-01), 397-401 0925-2738 50:4<397 2011 50 10858 https://doi.org/10.1007/s10858-011-9525-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9525-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kamiya Yukiko Okazaki Institute for Integrative Bioscience and Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, 444-8787, Okazaki, Aichi, Japan aut NATIONALLICENCE 700 1- Yamamoto Sayoko Okazaki Institute for Integrative Bioscience and Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, 444-8787, Okazaki, Aichi, Japan aut NATIONALLICENCE 700 1- Chiba Yasunori Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Tukuba Central 6, 1-1-1 Higashi, 305-8566, Tukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Jigami Yoshifumi Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology, Tukuba Central 6, 1-1-1 Higashi, 305-8566, Tukuba, Ibaraki, Japan aut NATIONALLICENCE 700 1- Kato Koichi Okazaki Institute for Integrative Bioscience and Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, 444-8787, Okazaki, Aichi, Japan aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 50/4(2011-08-01), 397-401 0925-2738 50:4<397 2011 50 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer