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

Evgeny Vinogradov; Bent O. Petersen; Jens Øllgaard Duus

doi:10.1016/S0008-6215(99)00332-8

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

Evgeny Vinogradov, Bent O. Petersen, Jens Øllgaard Duus

Carlsberg Research Center

Research output: Contribution to journal › Journal article › Research › peer-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 language	English
Journal	Carbohydrate Research
Volume	325
Issue number	3
Pages (from-to)	216-221
ISSN	0008-6215
DOIs	https://doi.org/10.1016/S0008-6215(99)00332-8
Publication status	Published - 2000
Externally published	Yes

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

Access to Document

10.1016/S0008-6215(99)00332-8

OpenUrl availability

Full text

Cite this

@article{a4b386fa5e2f4e0497029838251338e2,

title = "Isolation and characterization of non-labeled and C-13-labeled mannans from Pichia pastoris yeast",

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.",

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",

author = "Evgeny Vinogradov and Petersen, {Bent O.} and Duus, {Jens {\O}llgaard}",

year = "2000",

doi = "10.1016/S0008-6215(99)00332-8",

language = "English",

volume = "325",

pages = "216--221",

journal = "Carbohydrate Research",

issn = "0008-6215",

publisher = "Pergamon Press",

number = "3",

}

TY - JOUR

T1 - Isolation and characterization of non-labeled and C-13-labeled mannans from Pichia pastoris yeast

AU - Vinogradov, Evgeny

AU - Petersen, Bent O.

AU - Duus, Jens Øllgaard

PY - 2000

Y1 - 2000

N2 - 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.

AB - 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.

KW - Carbohydrate Sequence

KW - Carbon Isotopes

KW - Chromatography, DEAE-Cellulose

KW - Cloning, Molecular

KW - DNA, Complementary

KW - Magnetic Resonance Spectroscopy

KW - Mannans

KW - Molecular Sequence Data

KW - Pichia

KW - Polysaccharides

KW - carbon 13

KW - glucose

KW - mannan

KW - methanol

KW - oligosaccharide

KW - carbon nuclear magnetic resonance

KW - chemical structure

KW - controlled study

KW - DNA modification

KW - isolation procedure

KW - Pichia pastoris

KW - priority journal

KW - proton nuclear magnetic resonance

KW - short survey

KW - yeast

KW - 13C labeled

KW - Mannan

KW - Yeast

KW - BIOCHEMISTRY

KW - CHEMISTRY,

KW - SPIN-COUPLING CONSTANTS

KW - NMR-SPECTROSCOPY

KW - OLIGOSACCHARIDES

KW - ACETOLYSIS

KW - MAGNITUDE

KW - SIGN

KW - C-13 labeled

KW - Fungi Plantae (Fungi, Microorganisms, Nonvascular Plants, Plants) - Ascomycetes [15100] Pichia pastoris yeast

KW - mannans 9036-88-8 carbon-13 labeled, characterization, isolation, non-labeled

KW - 06504, Radiation biology - Radiation and isotope techniques

KW - 10058, Biochemistry methods - Carbohydrates

KW - 10068, Biochemistry studies - Carbohydrates

KW - 10504, Biophysics - Methods and techniques

KW - 51522, Plant physiology - Chemical constituents

KW - Bruker DRX 600 spectrometer Bruker, laboratory equipment

KW - gel filtration filtration techniques, isolation method

KW - NMR spectroscopy: analytical method, spectroscopic techniques CB

KW - Varian Unity Inova 500 spectrometer Varian, laboratory equipment

KW - Varian Unity Inova 750 spectrometer Varian, laboratory equipment

KW - Biochemistry and Molecular Biophysics

KW - Methods and Techniques

U2 - 10.1016/S0008-6215(99)00332-8

DO - 10.1016/S0008-6215(99)00332-8

M3 - Journal article

SN - 0008-6215

VL - 325

SP - 216

EP - 221

JO - Carbohydrate Research

JF - Carbohydrate Research

IS - 3

ER -

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

Abstract

Keywords

Access to Document

OpenUrl availability

Fingerprint

Cite this