Isolation and characterization of non-labeled and C-13-labeled mannans from Pichia pastoris yeast

Evgeny Vinogradov, Bent O. Petersen, Jens Øllgaard Duus

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Mannans from genetically modified Pichia pastoris yeast, used for overproduction of neural cell adhesion molecule protein, grown on normal media or on uniformly C-13-labeled glucose and methanol, were isolated and characterized by high-field (750 MHz) NMR spectroscopy. Fully C-13-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-(l-->6)-linked mannopyranosyl residues, substituted at O-2 with alpha-mannopyranosyl or 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. (C) 2000 Elsevier Science Ltd. All rights reserved.
Original languageEnglish
JournalCARBOHYDRATE RESEARCH
Volume325
Issue number3
Pages (from-to)216-221
ISSN0008-6215
DOIs
Publication statusPublished - 2000
Externally publishedYes

Keywords

  • Carbohydrate Sequence
  • Carbon Isotopes
  • Chromatography, DEAE-Cellulose
  • Cloning, Molecular
  • DNA, Complementary
  • Magnetic Resonance Spectroscopy
  • Mannans
  • Molecular Sequence Data
  • Pichia
  • Polysaccharides
  • carbon 13
  • glucose
  • mannan
  • methanol
  • oligosaccharide
  • carbon nuclear magnetic resonance
  • chemical structure
  • controlled study
  • DNA modification
  • isolation procedure
  • Pichia pastoris
  • priority journal
  • proton nuclear magnetic resonance
  • short survey
  • yeast
  • 13C labeled
  • Mannan
  • Yeast
  • BIOCHEMISTRY
  • CHEMISTRY,
  • SPIN-COUPLING CONSTANTS
  • NMR-SPECTROSCOPY
  • OLIGOSACCHARIDES
  • ACETOLYSIS
  • MAGNITUDE
  • SIGN
  • C-13 labeled
  • Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Ascomycetes [15100] Pichia pastoris yeast
  • mannans 9036-88-8 carbon-13 labeled, characterization, isolation, non-labeled
  • 06504, Radiation biology - Radiation and isotope techniques
  • 10058, Biochemistry methods - Carbohydrates
  • 10068, Biochemistry studies - Carbohydrates
  • 10504, Biophysics - Methods and techniques
  • 51522, Plant physiology - Chemical constituents
  • Bruker DRX 600 spectrometer Bruker, laboratory equipment
  • gel filtration filtration techniques, isolation method
  • NMR spectroscopy: analytical method, spectroscopic techniques CB
  • Varian Unity Inova 500 spectrometer Varian, laboratory equipment
  • Varian Unity Inova 750 spectrometer Varian, laboratory equipment
  • Biochemistry and Molecular Biophysics
  • Methods and Techniques

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