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Isolation and characterization of non-labeled and 13C-labeled mannans from Pichia pastoris yeast.
Carbohydr Res. 2000 Apr 20; 325(3):216-21.CR

Abstract

Mannans from genetically modified Pichia pastoris yeast, used for overproduction of neural cell adhesion molecule protein, grown on normal media or on uniformly 13C-labeled glucose and methanol, were isolated and characterized by high-field (750 MHz) NMR spectroscopy. Fully 13C-labeled oligosaccharide fragments were prepared from mannans by acetolysis. According to the data obtained, the mannan is made up of a main chain of alpha-(1-->6)-linked mannopyranosyl residues, substituted at 0-2 with alpha-mannopyranosyl or a alpha-D-Manp-(1-->2)-beta-D-Manp-(1-->2)-beta-D-Manp-(1-->2)-alpha-D-Manp- group, and with much lower content of substitution with beta-D-Manp-(1-->2)-alpha-D-Manp-. A fraction of these oligosaccharide side chains is again substituted with alpha-D-Glcp or alpha-D-GlcpNAc through a phosphodiester linkage to the 6 position of the first mannopyranosyl residue. Improved conditions of acetolysis, cleaving all alpha-(1-->6) linkages, but not beta-mannoside linkages, are proposed.

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Authors+Show Affiliations

Vinogradov E
Department of Chemistry, Carlsberg Laboratory, Copenhagen, Denmark.
Petersen BO
No affiliation info available
Duus JO
No affiliation info available

MeSH

Carbohydrate SequenceCarbon IsotopesChromatography, DEAE-CelluloseCloning, MolecularDNA, ComplementaryMagnetic Resonance SpectroscopyMannansMolecular Sequence DataPichiaPolysaccharides

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10795813

Citation

Vinogradov, E, et al. "Isolation and Characterization of Non-labeled and 13C-labeled Mannans From Pichia Pastoris Yeast." Carbohydrate Research, vol. 325, no. 3, 2000, pp. 216-21.
Vinogradov E, Petersen BO, Duus JO. Isolation and characterization of non-labeled and 13C-labeled mannans from Pichia pastoris yeast. Carbohydr Res. 2000;325(3):216-21.
Vinogradov, E., Petersen, B. O., & Duus, J. O. (2000). Isolation and characterization of non-labeled and 13C-labeled mannans from Pichia pastoris yeast. Carbohydrate Research, 325(3), 216-21.
Vinogradov E, Petersen BO, Duus JO. Isolation and Characterization of Non-labeled and 13C-labeled Mannans From Pichia Pastoris Yeast. Carbohydr Res. 2000 Apr 20;325(3):216-21. PubMed PMID: 10795813.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Isolation and characterization of non-labeled and 13C-labeled mannans from Pichia pastoris yeast. AU - Vinogradov,E, AU - Petersen,B O, AU - Duus,J O, PY - 2000/5/5/pubmed PY - 2000/7/15/medline PY - 2000/5/5/entrez SP - 216 EP - 21 JF - Carbohydrate research JO - Carbohydr Res VL - 325 IS - 3 N2 - Mannans from genetically modified Pichia pastoris yeast, used for overproduction of neural cell adhesion molecule protein, grown on normal media or on uniformly 13C-labeled glucose and methanol, were isolated and characterized by high-field (750 MHz) NMR spectroscopy. Fully 13C-labeled oligosaccharide fragments were prepared from mannans by acetolysis. According to the data obtained, the mannan is made up of a main chain of alpha-(1-->6)-linked mannopyranosyl residues, substituted at 0-2 with alpha-mannopyranosyl or a alpha-D-Manp-(1-->2)-beta-D-Manp-(1-->2)-beta-D-Manp-(1-->2)-alpha-D-Manp- group, and with much lower content of substitution with beta-D-Manp-(1-->2)-alpha-D-Manp-. A fraction of these oligosaccharide side chains is again substituted with alpha-D-Glcp or alpha-D-GlcpNAc through a phosphodiester linkage to the 6 position of the first mannopyranosyl residue. Improved conditions of acetolysis, cleaving all alpha-(1-->6) linkages, but not beta-mannoside linkages, are proposed. SN - 0008-6215 UR - https://www.unboundmedicine.com/medline/citation/10795813/Isolation_and_characterization_of_non_labeled_and_13C_labeled_mannans_from_Pichia_pastoris_yeast_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0008-6215(99)00332-8 DB - PRIME DP - Unbound Medicine ER -