Extracellular polysaccharides from suspension cultures of Nicotiana plumbaginifolia

Nicotiana Plumbaginifolia ◽

Suspension Cultures ◽

Exchange Chromatography ◽

Arabinogalactan Protein ◽

Selective Precipitation ◽

The Individual

The soluble polymers secreted by cell-suspension cultures of Nicotiana plumbaginifolia contained 78% carbohydrate, 6% protein and 4% inorganic material. The extracellular polysaccharides were separated into three fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7 and the individual polysaccharides in each fraction were then isolated by selective precipitation and enzymic treatment. Monosaccharide and linkage compositions were determined for each polysaccharide after reduction of uronic acid residues and the degree of esterification of the various uronic acid residues in each polysaccharide was determined concurrently with the linkage types. Six components were identified: an arabinoxyloglucan (comprising 34% of the total polysaccharide) and a galactoglucomannan (15%) in the unbound neutral fraction, a type II arabinogalactan (an arabinogalactan-protein, 11%) and an acidic xylan (3%) in the first bound fraction, and an arabinoglucuronomannan (11%) and a galacturonan (26%) in the second bound fraction. © 1995.

Characterisation of extracellular polysaccharides from suspension cultures of members of the Poaceae

10.26686/wgtn.12597854.v1 ◽

2020 ◽

Author(s):

Ian Sims ◽

K Middleton ◽

AG Lane ◽

AJ Cairns ◽

A Bacic

Keyword(s):

Cultured Cells ◽

Oryza Sativa L ◽

Suspension Cultures ◽

Exchange Chromatography ◽

Phleum Pratense ◽

Arabinogalactan Protein ◽

Type Ii ◽

Hordeum Vulgare L ◽

Suspension Cultured Cells

Microscopic examination of suspension-cultured cells of Phleum pratense L., Panicum miliaceum L., Phalaris aquatica L. and Oryza sativa L. showed that they were comprised of numerous root primordia. Polysaccharides secreted by these suspension cultures contained glycosyl linkages consistent with the presence of high proportions of root mucilage-like polysaccharides. In contrast, suspension-cultured cells of Hordeum vulgare L. contained mostly undifferentiated cells more typical of plant cells in suspension culture. The polysaccharides secreted by H. vulgare cultures contained mostly linkages consistent with the presence of glucuronoarabinoxylan. The soluble polymers secreted by cell-suspension cultures of Phleum pratense contained 70% carbohydrate, 14% protein and 6% inorganic material. The extracellular polysaccharides were separated into four fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7.0. From glycosyl-linkage analyses, five polysaccharides were identified: an arabinosylated xyloglucan (comprising 20% of the total polysaccharide), a glucomannan (6%), a type-II arabinogalactan (an arabinogalactan-protein; 7%), an acidic xylan (3%), and a root-slime-like polysaccharide, which contained features of type-II arabinogalactans and glucuronomannans (65%).

Characterisation of extracellular polysaccharides from suspension cultures of members of the Poaceae

10.26686/wgtn.12597854 ◽

2020 ◽

Author(s):

Ian Sims ◽

K Middleton ◽

AG Lane ◽

AJ Cairns ◽

A Bacic

Keyword(s):

Cultured Cells ◽

Oryza Sativa L ◽

Suspension Cultures ◽

Exchange Chromatography ◽

Phleum Pratense ◽

Arabinogalactan Protein ◽

Type Ii ◽

Hordeum Vulgare L ◽

Suspension Cultured Cells

Microscopic examination of suspension-cultured cells of Phleum pratense L., Panicum miliaceum L., Phalaris aquatica L. and Oryza sativa L. showed that they were comprised of numerous root primordia. Polysaccharides secreted by these suspension cultures contained glycosyl linkages consistent with the presence of high proportions of root mucilage-like polysaccharides. In contrast, suspension-cultured cells of Hordeum vulgare L. contained mostly undifferentiated cells more typical of plant cells in suspension culture. The polysaccharides secreted by H. vulgare cultures contained mostly linkages consistent with the presence of glucuronoarabinoxylan. The soluble polymers secreted by cell-suspension cultures of Phleum pratense contained 70% carbohydrate, 14% protein and 6% inorganic material. The extracellular polysaccharides were separated into four fractions by anion-exchange chromatography using a gradient of imidazole-HCl at pH 7.0. From glycosyl-linkage analyses, five polysaccharides were identified: an arabinosylated xyloglucan (comprising 20% of the total polysaccharide), a glucomannan (6%), a type-II arabinogalactan (an arabinogalactan-protein; 7%), an acidic xylan (3%), and a root-slime-like polysaccharide, which contained features of type-II arabinogalactans and glucuronomannans (65%).

Characterization of extracellular polysaccharides from suspension cultures of apple (Malus domestica)

10.26686/wgtn.12597845.v1 ◽

2020 ◽

Author(s):

S Reid ◽

Ian Sims ◽

LD Melton ◽

AM Gane

Keyword(s):

Monosaccharide Composition ◽

Exchange Chromatography ◽

Neutral Fraction ◽

Neutral Sugar ◽

Diffusion Test ◽

Selective Precipitation ◽

Acidic Fraction

The polymers secreted by suspension-cultured apple cells were composed of 85% carbohydrate (76% neutral sugar and 9% uronic acid) and 15% w/w protein. The extracellular polysaccharides (ECPs) contain 23% XG and 59% AGPs. The monosaccharide composition of the ECPs consisted of Gal, Ara, Glc and Xyl, with smaller amounts of Rha, Fuc and Man. Fractionation of the ECPs by anion-exchange chromatography yielded an unbound neutral fraction and a bound acidic fraction. Monosaccharide and linkage compositions of each fraction were determined. The neutral fraction (48% recovered carbohydrate) was composed of xyloglucan (XG;>90 mol%) which was purified by selective precipitation with Fehling's solution to yield pure XG. The purified XG had a Glc:Xyl:Gal:Fuc ratio of 4.0:2.5:0.8:0.5; the XG was not O-acetylated. The structure of the secreted XG was similar to that extracted from apple-pomace. The acidic fraction (52% recovered carbohydrate) was composed primarily of arabinogalactan-proteins (AGPs) as detected by the β-glucosyl Yariv diffusion test. The AGP had a Gal:Ara ratio of 1.3: 1.0. Minor amounts of arabinan, xylan and mannan were also detected in the ECPs. This study is the first examination of the polysaccharides secreted by apple cells grown in suspension culture.

Characterization of extracellular polysaccharides from suspension cultures of apple (Malus domestica)

10.26686/wgtn.12597845 ◽

2020 ◽

Author(s):

S Reid ◽

Ian Sims ◽

LD Melton ◽

AM Gane

Keyword(s):

Monosaccharide Composition ◽

Exchange Chromatography ◽

Neutral Fraction ◽

Neutral Sugar ◽

Diffusion Test ◽

Selective Precipitation ◽

Acidic Fraction

The polymers secreted by suspension-cultured apple cells were composed of 85% carbohydrate (76% neutral sugar and 9% uronic acid) and 15% w/w protein. The extracellular polysaccharides (ECPs) contain 23% XG and 59% AGPs. The monosaccharide composition of the ECPs consisted of Gal, Ara, Glc and Xyl, with smaller amounts of Rha, Fuc and Man. Fractionation of the ECPs by anion-exchange chromatography yielded an unbound neutral fraction and a bound acidic fraction. Monosaccharide and linkage compositions of each fraction were determined. The neutral fraction (48% recovered carbohydrate) was composed of xyloglucan (XG;>90 mol%) which was purified by selective precipitation with Fehling's solution to yield pure XG. The purified XG had a Glc:Xyl:Gal:Fuc ratio of 4.0:2.5:0.8:0.5; the XG was not O-acetylated. The structure of the secreted XG was similar to that extracted from apple-pomace. The acidic fraction (52% recovered carbohydrate) was composed primarily of arabinogalactan-proteins (AGPs) as detected by the β-glucosyl Yariv diffusion test. The AGP had a Gal:Ara ratio of 1.3: 1.0. Minor amounts of arabinan, xylan and mannan were also detected in the ECPs. This study is the first examination of the polysaccharides secreted by apple cells grown in suspension culture.

Heterogeneity in the collagens extracted from human embryonic calvaria

Canadian Journal of Biochemistry and Cell Biology ◽

10.1139/o83-129 ◽

1983 ◽

Vol 61 (9) ◽

pp. 1012-1017 ◽

Cited By ~ 2

Author(s):

James R. A. Leushner

Keyword(s):

Type I Collagen ◽

Exchange Chromatography ◽

Exact Nature ◽

Type I ◽

Selective Precipitation ◽

Nacl Concentration ◽

Chain Composition ◽

The Individual ◽

Type V

Collagens were obtained from decalcified human embryonic calvaria by pepsin digestion. After removal of the type I collagen, the more soluble collagens were precipitated at 1.2 M NaCl (acid pH), followed by a selective precipitation step at neutral pH, using a NaCl concentration of 4.5 M. Analysis of this latter precipitate by polyacrylamide slab gel electrophoresis and ion-exchange chromatography revealed the presence of a heterogeneous group of proteins ranging in size from approximately 10 000 daltons to over 120 000 daltons. Proteolysis, as a source for these diverse components, was ruled out both by studies employing protease inhibitors and experiments employing thrombin which indicated that no helical denaturation had occurred during extraction. A comparison with standard collagen preparations suggested that the major bands present in these samples corresponded to the α1-, α2-, and α3- chains of type V collagen. The data also showed that the major helical organization of these chains was [α1(V)]2 α2(V). Data suggesting that proteolysis can occur during the chromatographic separation of the individual α-chains are presented. This proteolysis was sensitive to inhibitors and its possible role in modifying the chain composition of type from calvaria and other tissues is discussed. Unique cyanogen bromide peptides distinguishable from those of type I and type V were present in these precipitates and suggests the presence of novel collagen types. The small amounts of these collagens precluded a determination of the exact nature of these components.

The compositional analysis of bacterial extracellular polysaccharides by high-performance anion-exchange chromatography

Analytical Biochemistry ◽

10.1016/0003-2697(91)90270-4 ◽

1991 ◽

Vol 199 (1) ◽

pp. 68-74 ◽

Cited By ~ 51

Author(s):

Anthony J. Clarke ◽

Vivian Sarabia ◽

Wendy Keenleyside ◽

P. Ronald MacLachlan ◽

Chris Whitfield

Keyword(s):

Anion Exchange ◽

High Performance ◽

Compositional Analysis ◽

Exchange Chromatography ◽

Extracellular Polysaccharides

Enzymatic Studies on the Reversible Synthesis of Nicotinic Acid-N-glucoside in Heterotrophic Parsley Cell Suspension Cultures

Zeitschrift für Naturforschung C ◽

10.1515/znc-1988-11-1208 ◽

1988 ◽

Vol 43 (11-12) ◽

pp. 835-842 ◽

Cited By ~ 13

Author(s):

Barbara Upmeier ◽

Jürgen E. Thomzik ◽

Wolfgang Barz

Keyword(s):

Nicotinic Acid ◽

Cell Suspension ◽

Gel Filtration ◽

Cell Suspension Cultures ◽

Suspension Cultures ◽

Exchange Chromatography ◽

High Group ◽

Ph Optimum ◽

Glucose Molecule ◽

Transfer Potential

Abstract A soluble enzyme catalyzing the transfer of the glucose moiety from UDP-glucose to the nitrogen atom of nicotinic acid was detected in protein preparations from heterotrophic cell suspension cultures of parsley (Petroselinum hortense Hoffm.). Enzyme activity was enriched 22-fold by ammonium sulfate precipitation, gel filtration and ion exchange chromatography. The UDP-glucose : nicotinic acid-N-glucosyltransferase showed a pH-optimum at pH 7.8-8.2 and a temperature optimum at 30 °C. The apparent KM values were determined to be 170 μM for nicotinic acid and 1.2 mM for the cosubstrate. The native enzyme had a molecular mass of about 46 kDa. The glucosyltransferase reaction was shown to be reversible. The transfer of the glucose molecule from nicotinic acid-N-glucoside to uridinediphosphate yielding uridinediphosphoglucose and nicotinic acid could be demonstrated indicating that nicotinic acid-N-glucoside has a high group-transfer potential.