A rapid and sensitive method for the analysis of carbohydrate components in glycoproteins using gas-liquid chromatography

doi:10.1016/0003-2697(82)90455-9

Analytical Biochemistry

Volume 123, Issue 2, 1 July 1982, Pages 336-341

https://doi.org/10.1016/0003-2697(82)90455-9 Get rights and content

Abstract

A rapid and simple gas-liquid chromatographic method for the determination of subnanomolar amounts of carbohydrates derived from glycoproteins is described. The procedure involves methanolysis in the presence of methyl acetate followed by removal of hydrogen chloride by coevaporation with t-butyl alcohol and trimethylsilylation. The method is also applicable to samples containing uronic acids and lipids.

References (25)

A.K. Allen et al.
FEBS Lett
(1975)
T. Kato et al.
Anal. Biochem
(1980)
W.F. Alpenfels
Anal. Biochem
(1981)
M. D'Amboise et al.
Carbohydr. Res
(1980)
J. Lehrfeld
Anal. Biochem
(1981)
T.P. Mawhinney et al.
Anal. Biochem
(1980)
R.R. Selvendran et al.
Anal. Biochem
(1979)
M. Yokota et al.
Carbohydr. Res
(1977)
J. Conchie
T. Bhatti et al.
Biochim. Biophys. Acta
(1970)

S.J. Rickert et al.

J. Chromatogr

(1978)

G.A. Levvy et al.

Biochim. Biophys. Acta

(1970)

Cited by (292)

Characterization of an exopolysaccharide (EPS-3A) produced by Streptococcus thermophilus ZJUIDS-2-01 isolated from traditional yak yogurt
2021, International Journal of Biological Macromolecules
Citation Excerpt :
The spectrum was recorded in 4000–400 cm−1 with 16 scans and analyzed by the FT-IR 650 software (Gangdong, Tianjin, China). The methylation of EPS-3A was performed as described [35,36]. Briefly, 3 mg of EPS-3A were dissolved in DMSO (5 mL) by sonication, and 40 mg NaOH (fine powder) were added.
Yak yogurt, one of the naturally fermented dairy products prepared by local herdsmen in the Qinghai-Tibet Plateau, contains a diverse array of microorganisms. We isolated and identified a novel Streptococcus thermophilus strain, ZJUIDS-2-01, from the traditional yak yogurt. We further purified and carried out detailed structural, physiochemical, and bioactivity studies of an exopolysaccharide (EPS-3A) produced by S. thermophilus ZJUIDS-2-01. The weight-average molecular weight (Mw) of EPS-3A was estimated to be 1.38 × 10⁶ Da by High-Performance Gel Permeation Chromatography (HPGPC). The monosaccharide analysis established its composition to be glucose, galactose, N-acetyl-D-galactosamine, and rhamnose in a ratio of 5.2:2.5:6.4:1.0. The molecular structure of EPS-3A was determined by the combination of permethylation analysis, FT-IR, and NMR spectroscopic techniques. The ζ-potential measurements indicated that EPS-3A had a pKa value of ~4.40. The DSC yielded a melting point (T_m) of 80.4 °C and enthalpy change (ΔH) of 578 J/g for EPS-3A, comparable to those of the xanthan gum (XG), a commercial EPS. EPS-3A exhibited better O/W emulsion stability and flocculating capacity than XG. Furthermore, it also demonstrated similar antioxidant activity to XG and promising in vitro antibacterial properties. This work evidenced that EPS-3A derived from S. thermophilus ZJUIDS-2-01 holds the potential for food and industrial applications.
Isolation and characterization of halophilic bacteria producing exopolymers with emulsifying and antioxidant activities
2018, Biocatalysis and Agricultural Biotechnology
Halophilic bacteria are considered a great source of new strains producing novel exopolymers with functional properties. In this work we isolated ten halophilic strains producing exopolymers from different hypersaline environments in Morocco. Phenotypic characterization showed that the strains were moderately halophilic, mesophilic and neutrophilic with the ability to produce some hydrolytic enzymes. Strains identification based on 16S rRNA gene sequences comparison showed that nine strains, designed as N1, N2, N5, N7, N8, N9, N10, N11 and N12 belong to Halomonas genus and one strain, designed as N4, to Marinobacter genus. The majority of the strains showed high levels of exopolymer production. The study of emulsifying and antioxidant activities revealed that all the polymers have an interesting emulsifying and antioxidant activities with the polymer from Marinobacter sp. N4 forming the highest and most stable emulsions and exhibiting the best antioxidant activity in comparison with other exopolymers produced by Halomonas strains. The obtained results demonstrate the great potential of exopolymers from halophilic bacteria to be applied as emulsifying and antioxidant agents in food, cosmetics and oil industries.
Structure of surface polysaccharides from Aeromonas sp. AMG272, a plant-growth promoting rhizobacterium isolated from rice rhizosphere
2018, Carbohydrate Research
Aeromonas sp. AMG272 is a Gram-negative bacterium that has been isolated from agricultural soil and studied for its plant growth-promoting activities. Structures of the O-specific polysaccharide chain of the AMG272 lipopolysaccharide and its capsular polysaccharide were elucidated using GLC-MS and NMR spectroscopy. The structure of the O-specific polysaccharide, →4)-α-l-Rhap-(1 → 3)-β-d-GlcpNAc-(1→, has been found in other Aeromonas strains and related bacteria, whereas the structure of the capsular polysaccharide has not been reported before:
→6)[β-d-Fucp3NAc4Ac-(1 → 3)]-α-d-GlcpNAc-(1 → 4)-α-d-Galp-(1 → 3)-α-d-GalpNAc-(1 → 4)-α-d-Galp-(1 → .
Conformational studies and molecular characterization of the polysaccharide kefiran
2018, Food Hydrocolloids
Kefiran was produced from kefir grains, on a semi-industrial scale, using UHT skimmed milk as a raw material. The conformational characteristics of the polysaccharide were assessed by means of a series of techniques such as capillary viscometry, size-exclusion chromatography coupled to multi-angle laser light scattering, static light scattering, dynamic light scattering and zeta-potential measurements. The results indicated that kefiran behaved as a weakly charged polysaccharide with a having, possibly, a random coil conformation in aqueous solution. The weight average molecular weight was 6.71 × 10⁵ Da, whereas the mean square radius of gyration was 43.2 nm. The effect of a series of solutes in various concentrations such as salts, organic acids, KOH, sugars, urea, ethanol, Na-caseinate and whey protein concentrates on the flow behavior of kefiran in aqueous dilute solutions was assessed by hydrodynamic, zeta potential and dynamic light scattering measurements. The results are discussed in the light of conformational changes observed.
Characterization of haloglycan, an exopolysaccharide produced by Halomonas stenophila HK30
2015, International Journal of Biological Macromolecules
We have conducted a thorough study of the exopolysaccharide (EPS) produced by strain HK30 of Halomonas stenophila, which we have named haloglycan. This strain was chosen during an ongoing research programme aimed at finding novel exopolysaccharide-producing halophilic bacteria in unexplored hypersaline habitats. Strain HK30 was isolated from a saline-wetland in Brikcha (Morocco) and identified as belonging to the species H. stenophila. It produced EPS mainly during the exponential growth phase and to a lesser extent during the stationary phase. Culture parameters influenced both bacterial growth and EPS production, EPS yield always being directly related to the quantity of biomass. Under optimum culture conditions, strain HK30 produced 3.89 g of EPS per litre of medium. The polymer was a sulphated heteropolysaccharide composed of two fractions, with molecular masses of 8.2 × 10⁴ and 1.4 × 10⁶. The crude EPS contained 44 ± 0.1% w/w carbohydrates and the following monosaccharide composition: glucose (24 ± 1.73), glucuronic acid (7.5 ± 0.37), mannose (5.5 ± 0.17), fucose (4.5 ± 0.36), galactose (1.2 ± 0.17) and rhamnose (1 ± 0.05) (%, w/w). It produced solutions of high viscosity and pseudoplastic behaviour that showed interesting flocculating and emulsifying activities and was also involved in forming biofilm.
The Yariv reagent: Behaviour in different solvents and interaction with a gum arabic arabinogalactanprotein
2014, Carbohydrate Polymers
The β-d-Glc Yariv reagent is frequently used to isolate and to study the structure of arabinogalactan-proteins with the arabinogalactan type II structure. The present paper describes the aggregation features of the Yariv reagent in water, salt solutions and in organic solvents as determined by NMR, absorption spectroscopy and light scattering experiments. The results indicate that in water the Yariv reagent forms aggregates of up to 300 units and in 1% aqueous NaCl the degree of aggregation is approx. 150. The aggregates are formed both by H-bonds and hydrophobic interactions, the former appearing to be of most importance in water.
The interaction between the Yariv reagent and an AGP fraction from gum arabic, showed a degree of aggregation of the Yariv reagent when using 1% NaCl to be of approx. 150 units, whereas disruption of the aggregate took place in 10% NaCl with an aggregation number of approx. 100.
Partial acid hydrolysis of an AGP from gum Arabic (Acacia Senegal) and analyses of the linkage types remaining indicated that a certain length of (1 → 3)-β-linked galactose units was necessary for binding between the Yariv reagent and the AGP. This is in accordance to what also was recently observed by Kitazawa et al. (2013).

View all citing articles on Scopus

^☆: This work was supported by Medical Research Council Grant G80/0633/7CA.

View full text

A rapid and sensitive method for the analysis of carbohydrate components in glycoproteins using gas-liquid chromatography☆

Abstract

FEBS Lett

Anal. Biochem

Anal. Biochem

Carbohydr. Res

Anal. Biochem

Anal. Biochem

Anal. Biochem

Carbohydr. Res

Biochim. Biophys. Acta

J. Chromatogr

Biochim. Biophys. Acta