Characterisation of extracellular polysaccharides produced by Crypthecodinium cohnii

The valuable polyunsaturated fatty acid, docosahexaenoic acid, can be produced by cultivation of the heterotrophic microalga, Crypthecodinium cohnii. During batch growth of C. cohnii on glucose, sea salt and yeast extract for 5 days, so far unreported extracellular polysaccharides were produced. These caused an increased viscosity and a strong drop in the maximum oxygen transfer. The viscosity increased most markedly as cells entered the stationary phase. The polysaccharides varied in size (from 6 kDa to >1,660 kDa) and monomer distribution. A high molecular mass fraction (from 100 kDa to >1,660 kDa) and a medium molecular mass fraction (6-48 kDa) were prepared. The high molecular mass fraction contained (on a molar basis) 71.7% glucose, 13.1% galactose and 3.8% mannose, whereas the medium molecular mass fraction contained 37.7% glucose, 19.8% galactose and 28.1% mannose. Other monomers present in both fractions were fucose, uronic acid and xylose. Monomers were coupled mainly via alpha-(1-3) links. Increased viscosity due to polysaccharide production complicates the development of commercial, high cell-density processes for the production of docosahexaenoic acid.     

Preliminary studies on the composition and rheological properties of the extracellular polysaccharide synthesized by Pseudomonas PBI (NCIB 11264).

A microorganism isolated from a carbohydrate-rich industrial effluent synthesized an exocellular slime polysaccharide composed of glucose and galactose in a molar ratio of 7.45 +/- 0.68 : 1, and two non-carbohydrate substituents acetate (3--4%) and pyruvate (5--9%). Contamination by rhamnose and mannose was detectable in crude polysaccharide samples. Solutions of the polysaccharide were pseudoplastic, but not thixotropic, and formed gels in the presence of certain trivalent cations.     

Preliminary studies on the composition and rheological properties of the extracellular polysaccharide synthesized by pseudomonas PB1 (NCIB 11264)

A microorganism isolated from a carbohydrate-rich industrial effluent synthesized an exocellular slime polysaccharide composed of glucose and galactose in a molar ratio of 7.45 ± 0.68: 1, and two non-carbohydrate substituents acetate (3–4%) and pyruvate (5–9%). Contamination by rhamnose and mannose was detectable in crude polysaccharide samples. Solutions of the polysaccharide were pseudoplastic, but not thixotropic, and formed gels in the presence of certain trivalent cations.     

灵芝液态发酵高产胞外多糖菌株筛选及多糖特性分析

对10株灵芝菌株发酵菌丝体生物量、胞内多糖含量、胞外粗多糖得率及其多糖含量和单糖组成特征进行了系统分析。结果表明,随着培养时间的延长,10株菌株的生物量均有不同程度增加,但不同菌株胞内多糖含量和胞外粗多糖得率变化趋势有所不同。在培养至7d时,G0023和G0160菌株胞外粗多糖得率均较高,分别为3.02g/L和3.14g/L,其多糖含量分别达到了84.11%和91.03%,可作为发酵高产胞外多糖的优良菌株。分级醇沉胞外多糖分析结果表明,各菌株胞外液20%乙醇沉淀所得20E组分的得率和多糖含量均高于50%乙醇沉淀所得的50E组分,说明胞外液中主要以大分子量多糖为主。20E主要由葡萄糖组成,含有少量木糖和甘露糖;50E主要由葡萄糖和甘露糖组成,含有少量木糖和半乳糖。胞外液表观粘度随剪切速率变化曲线分析结果显示,各菌株胞外液均呈现剪切变稀非牛顿流体特性,其表观粘度大小与菌株对应20E组分的得率及多糖含量呈正相关。     

Polysaccharides of Type 6 Klebsiella

Water-extractable type 6 Klebsiella antigens were separated into a type 6-specific acidic polysaccharide and a neutral polysaccharide. The neutral polymer was devoid of type 6 activity although it was serologically active. The type 6-specific polymer contained fucose, glucose, and mannose, and pyruvic, galacturonic, and possibly glucuronic acids. The neutral polymer contained glucose, galactose, and mannose.     

Chemical characterization of the hyphal walls of the basidiomyceteArmillaria mellea

Purified hyphal walls ofArmillaria mellea were analyzed and shown to contain glucose (59.0%), mannose (6.9%), galactose (4.2%), xylose (0.2%), ribose (traces), glucosamine (6.7%), protein (10.5%), and some lipid material (8.0%). The mucilaginous surface polysaccharide (fraction I) consisted mainly of (1–3), (1–4), and (1–6)β-glucan chains with mannose, galactose, xylose, and ribose. Fraction II was made mostly of (1–3)-linkedα-glucan. Fraction III contained mainly (1–3)-linkedβ-glucan with small amounts of mannose and galactose, and fraction IV was an alkali-insolubleβ(1–3)-glucan in close association with chitin.     

Nuclear columns: labeling and release of virus specific RNA by nuclei of DNA-virus (SV 40) infected cells

Lipopolysaccharide and an acidic polysaccharide were extracted with phenol-water from a rough strain of $\text{Escherichia coli}$ (LP1092). The polysaccharide portion of lipopolysaccharide contained galactose, glucose, L-glycero-D-mannoheptose, small amounts of mannose and an unusually high proportion of 3-deoxy-D-manno-octulosonic acid; this polysaccharide was shown to represent the complete $\text{coli}$ R2 core. The acidic polysaccharide, which functioned as a K antigen, contained large amounts of a 2-keto-3-deoxy sugar acid. On colorimetric and chromatographic evidence this acid appeared to be 3-deoxy-D-manno-octulosonic acid.     

Chemistry of the polysaccharides of the diatom Coscinodiscus nobilis

The extracellular polysaccharide of Coscinodiscus nobilis, a member of the Coscinodiscaceae, contains a highly branched heteropolysaccharide(s) containing fucose, rhamnose, mannose, d-glucose, xylose, d-glucuronic acid, galactose (trace) and half ester sulphate. The positions of linkages between the monosaccharides have been established and evidence for the linkages between d-glucuronic acid and monosaccharides was obtained. The extracellular polysaccharide contained also a chrysolaminaran, but this may have been derived from dead cells. Fucose and mannose occur also in a separate polymer. The diatom contained polysaccharide material consisting of glucose, mannose, fucose and uronic acid residues.     

Sugar compositions of extra- and intracellular polysaccharides produced by candida lipolytica (4124) grown on n-hexadecane

The free monosaccharide content of C. lipolytica (strain 4 124) cells grown on n-hexadecane was identified and found to be only glucose. The chromatographic analysis of the hydrolysate of intracellular cell wall polysaccharides indicated the presence of glucose: mannose: galactose: xylose in a ratio of 1 : 1.32 : 1.07 : 0.35. Paper and dise electrophoresis of extracellular polysaccharid from the culture broth was found to be heterogeneous. Ethanol fractionation separated it to a major component F (I) 81.99% and a minor one F (II) 13.04%. Analysis of the major fraction showed that it consisted of galactose and mannose only while the minor polysaccharide consisted of galactose, glucose and mannose. Thus it was concluded that the predominant sugar in both extracellular and intracellular polysaccharides was mannose. Xylose was detected in the intracellular polysaccharide only.     

Extracellular Polysaccharide Formed by Pseudomonas hydrogenovora in Autotrophic Culture and Its Physiological Activities

Gram negative hydrogen bacterium Pseudomonas hydrogenovora was found to excrete an anthrone-H₂SO₄ positive viscous polysaccharide. About 12 g/liter of the polysaccharide was produced autotrophically on gaseous hydrogen at the stationary phase of growth. Biosynthesis of the polysaccharide occurred under nitrogen-deficient condition. Its elementary composition was C: 39.29%, H: 6.23%, O: 49.67%, N: 0.21% and ash 4.6%. The polysaccharide contained galactose, glucose, mannose and rhamnose as its main components. The polysaccharide had anti-tobacco mosaic virus and anti-tumor activities.     

Plant suspension culture media macromolecules-pectic substances, protein, and peroxidase

Macromolecules secreted into the media by a nondifferentiating suspension culture of tobacco cells were found to be composed of protein and polysaccharide, and to account for the viscosity of the media. The concentration, composition, and viscosity of these macromolecules changed significantly with the age of the culture and growth temperature. The concentration changed from 0.02 mg/ml in newly inoculated cultures to over 1 mg/ml in older cultures. The macromolecules contained from 6 to 18% protein and 3 to 4 μmoles hydroxy-proline/mg nitrogen, more than 20 times the level found in whole cells. The macromolecules contained 5 to 25% pectic substances whose carboxyl groups were either methyl esterified or combined with calcium. Arabinose, xylose, glucose, galactose, and mannose were identified in acid hydrolysates of the macromolecules. Peroxidase activity of the macromolecules increased as cultures became older. Peroxidase isoenzyme patterns changed with culture age and growth temperature. The relation of the macromolecules to cell walls and intercellular substances is discussed.     

Isolation and characterization of a new extracellular polysaccharide from an Acetobacter species

An expolysaccharide (EPS)-producing strain of the genus Acetobacter (named IFR 101) was isolated from a commercial vinegar acidifier. IFR 101 can be grown over the pH range 3.0–8.0 and the temperature range 12–35°C, and is ethanol-tolerant to 10% v/v. Neutral sugar analysis of the purified EPS indicates the presence of the sugars mannose, galactose and glucose. Methylation studies and NMR data suggest that the EPS may either be a complex branched polysaccharide or a mixture of simpler linear and branched structures. Aqueous preparations of IFR 101 exhibit reversible shear-thinning behaviour.     

Production of a New Acidic Polysaccharide,Succinoglucan by Alcaligenes faecalis var. myxogenes

A slimy non-spore-forming bacterium strain 10C3 isolated from soil was motile with peritrichous flagella and named Alcaligenes faecalis var. myxogenes. Studies were made on the conditions necessary for maximal production of a new acidic succinoglucan polysaccharide by this strain in shaken cultures. Much production was observed with sucrose, glucose, xylose, galactose, cellobiose, maltose, fructose, mannose and rhamnose. The yield was greatest with sucrose and decreased in order with the above sugars from about 36 to 23 per cent. The most suitable medium contained 4 per cent sugar, 0.5 per cent yeast extract and one per cent calcium carbonate in tap water. The optimum temperature was 28°C.     

The extracellular polysaccharides of Rhizobium japonicum: Compositional studies

The extracellular polysaccharides of seven strains of Rhizobium japonicum were investigated by using a gas-chromatographic scheme developed for determination of the various sugars present. These polysaccharides were more heterogeneous in their composition than those of any other species of Rhizobium yet examined. Five strains (1809, 110, 123, 127, and 709) produced polysaccharides containing the same constituents, although in varying relative amounts: glucose (36–44%), galactose (7–25%), mannose (18–20%), 4-O-methylgalactose (5–13%), galacturonic acid (12–16%), and acetyl groups (4–8%). The sugars of the polysaccharide of strain 1809 were all of the d series. These are the first bacterial polysaccharides reported to contain 4-O-methylgalactose and the first Rhizobium polysaccharides in which galacturonic acid has been found. In contrast to this, the polysaccharide of strain 129 consisted of glucose (7%), galactose (51%), mannose (5%), xylose (5%), glucuronic acid (5%), and pyruvic acid (2%). The polysaccharide of strain 711 contained glucose (34%), galactose (13%), mannose (27%), and pyruvic acid (6%).     

Extracellular polysaccharide production by aRhzobium sp. from root nodules ofDerris scandens

TheRhizobium sp. isolated from the root nodules of the leguminous climbing shrubDerris scandens produced a large amount of extracellular polysaccharides in a yeast extract—mannitol medium in the stationary phase of growth. The production was maximum when the medium was supplemented with mannitol (3%), (+)-biotin (3 mg/L) and KNO₃ (0.3%). The extracellular polysaccharides contained glucose, galactose and mannose. The possible role of the rhizobial extracellular polysaccharide is discussed.     

Polysaccharide and glycolipid composition in Tritrichomonas foetus

1. The polysaccharide and glycolipid composition in Tritrichomonas foetus was studied by paper, thin-layer and gas-liquid chromatographic analysis. 2. The carbohydrate components of the polysaccharide were glucose (47%), galactose (34%) and mannose (19%). N-acetylneuraminic acid was the sialic acid derivative characterized in the flagellate whole cells. 3. The sialic acid density was estimated as 2.7 x 10(7) residues/cell. 4. The long-chain base dihydrosphingosine, the carbohydrates galactose (67%), glucose (21%) and mannose (12%) as well as the fatty acids myristic (48%) and palmitic (52%) acids were characterized as components of the total glycolipids of T. foetus. 5. Total glycolipids were fractionated: a galactocerebroside and a ganglioside were identified.     

Extracellular polysaccharides of some basidiomycetes.

Culture filtrates of four basidiomycete fungi, Stereum strigoso-zonatum, Fomes australis, Trametes lilacinogilva and Polyporus tumulosus were fractionated and examined for polysaccharide content. Acid hydrolysis showed the presence of galactose, mannose, xylose, fucose and glucose. Their relative amounts were estimated by gas chromatography of the corresponding alditol acetates. Galactose and mannose were the major constituent sugars, amounting to more than 50% of the total. One of the polysaccharides, a fucogalactomannan elaborated by P. tumulosus, was isolated in a purified form. It was shown to have [alpha]D +42 degrees and contained galactose, mannose, fucose and xylose in the relative proportions 2 : 1 : 1 : 0-2.     

Production of exopolysaccharides from a thermophilic microorganism isolated from a marine hot spring in flegrean areas

A thermophilic strain isolated from sea sand at Maronti, near Sant' Angelo (Ischia), is described. The organism grows well at an optimal temperature of 60 °C at pH 7.0. The thermophilic bacterium, named strain 4004, produces an exocellular polysaccharide (EPS) in yields of 90 mg/l. The EPS fraction was produced with all substrates tested, although a higher yield was obtained with sucrose or trehalose as sole carbon source. During growth, the EPS content was proportional to the biomass. Three fractions (EPS1, EPS2, EPS3) were obtained after purification. Quantitative monosaccharide analysis of the EPSs revealed the presence of mannose:glucose:galactose in a relative ratio of 0.5:1.0:0.3 in EPS1, mannose:glucose:galactose in a relative ratio of 1.0:0.3:trace in EPS2, and galactose:mannose:glucosamine:arabinose in a relative ratio of 1.0:0.8:0.4:0.2 in EPS3. The average molecular mass of EPS3 was determined to be 1×10⁶ Da. From comparison of the chemical shift values in ¹H and ¹³C spectra, we conclude that EPS3 presents a pentasaccharide repeating unit. Electronic Publication     

Investigation of lipopolysaccharides from Sinorhizobium meliloti SKHM1-188 and two of its mutants with decreased nodulation competitiveness

A comparative study of the lipopolysaccharides (LPS) isolated from Sinorhizobium meliloti SKHM 1-188 and two its LPS-mutants (Th29 and Ts22) with sharply decreased nodulation competitiveness was conducted. Polyacrylamide gel electrophoresis with sodium dodecyl sulfate revealed two forms of LPS in all the three strains: a higher molecular-weight LPS1, containing O-polysaccharide (O-PS), and a and lower molecular-weight LPS2 without O-PS. However, the LPS1 content in mutants was significantly smaller than in the parent strain. The LPS of the strains studied contained glucose, galactose, mannose, xylose, three nonidentified sugars--X1 (TGlc 0.53), X2 (TGlc 0.47), and X3 (TGlc 0.43), glucosamine, and ethanolamine, while the LPS of S. meliloti SKHM1-188 additionally contained galactosamine, glucuronic and galacturonic acids, and 2-keto-3-deoxyoctulosonic acid (KDO), as well as fatty acids, such as 3-OH C14:0, 3-OH C15:0, 3-OH C16:0, 3-OH C18:0, nonidentified hydroxy X (T3-OH C14:0 1.33), C18:0, and unsaturated C18:1 fatty acids. The LPS of both mutants were similar in the component composition but differed from the LPS of the parent strain by a lower X2, X3, and 3-OH C 14:0 content and a higher KDO, C18:0, and hydroxy X content. The LPS of all the strains were subjected to mild hydrolysis with 1% acetic acid and fractionated on a column with Sephadex G-25. The higher molecular weight fractions (2500-4000 Da) contained a set of sugars typical of intact LPS and, supposedly, corresponded to the LPS polysaccharide portion (PS1). In the lower molecular weight fractions (600-770 Da, PS2), glucose and uronic acids were the major components; galactose, mannose, and X1 were present in smaller amounts. The PS1/PS2 ratio for the two mutants was significantly lower than for strain SKHM1-188. The data obtained show that the amount of O-PS-containing molecules (LPS1) in the heterogeneous lipopolysaccharide complex of the mutants was smaller than in the SKHM1-188 LPS; this increases the hydrophobicity of the cell surface of the mutant bacteria. This supposedly contributes to their nonspecific adhesion on the roots of the host plant, thus decreasing their nodulation competitiveness.     

Immunochemical studies of Rhizobium mutants

Summary It was shown in gel diffusion and in immunoelectrophoresis tests that somatic antigens of smooth cultures were more complex than those of rough mutants. Somatic antigens ofR. trifolii 24 contained glucuronic acid, glucosamine, galactose, glucose, mannose, xylose or fucose, ribose, and 2-keto-3-deoxyoctonate. Arabinose was identified in some preparations. The antigens of the smooth strain contained a large amount of rhamnose whereas those of the rough mutants were deficient in that sugar.