Lipopolysaccharides of the acidophilic heterotrophic bacteria Acidiphilium cryptum and Acidiphilium symbioticum

Abstract Lipopolysaccharides of Acidiphilium cryptum and Acidiphilium symbioticum , the acidophilic heterotrophs of sulfidic mine environments, contain rhamnose, glucose, galactose and mannose. Molar ratios of the neutral sugars differ in these two lipopolysaccharide preparations. 3-Deoxy-d-manno-2-octulosonic acid is present in both the lipopolysaccharides, and galactosamine was the only amino sugar detected. 3-Hydroxytetradecanoic acid was found to be the major fatty acid component along with minute amounts of decanoic and dodecanoic acids. Deoxycholate-PAGE electrophoresis revealed the presence of short sugar chains and unsubstituted cores. The implications of these findings are discussed in relation to acidiphilicity and systematics of Acidiphilium species.     

Biochemical studies on the cell wall lipopolysaccharides (O-antigens) of Vibrio cholerae 569 B (Inaba) and El-tor (Inaba).

Lipopolysaccharides were isolated from the cell walls of Vibrio cholerae 569 B (Inaba) and El-tor (Inaba). Chemical analysis revealed the presence of glucose, fructose, mannose, heptose, rhamnose, ethanolamine, fatty acids and glucosamine. The lipopolysaccharides do not contain 2-keto-3-deoxyoctonate, the typical linking sugar of polysaccharide and lipid moieties of enterobacterial lipopolysaccharides. Galactose, a typical core polysaccharide component of many gram-negative bacteria was also absent from lipopolysaccharides of these organisms. By hydrolysis in 1% acetic acid, the lipopolysaccharides have been separated into a polysaccharide part (degraded polysaccharide) and a lipid part (lipid A). Components of degraded polysaccharide and lipid A moiety were identified and determined. The lipid A fractions contained fatty acids, phosphorus and glucosamine. All the neutral sugars detected in lipopolysaccharides were shown to be the constituents of its polysaccharide moiety. The fatty acid analysis of lipopolysaccharide and lipid A showed the presence of both hydroxy and non hydroxy acids. They were different from those of lipids extracted from cell walls before the extraction of lipopolysaccharides. 3-Hydroxylauric and 3-hydroxymyristic acids predominated in lipopolysaccharide and lipid A of Vibrio cholerae and El-tor (Inaba).     

Chemical composition of the lipopolysaccharides of Rhodobacter sulfidophilus,Rhodopseudomonas acidophila,and Rhodopseudomonas blastica

The lipopolysaccharides of Rhodobacter sulfidophilus and the two budding species Rhodopseudomonas acidophila and Rhodopseudomonas blastica were isolated and chemically analyzed. The all have a lipid A backbone structure with glucosamine as the only amino sugar. The lipid A's of Rb. sulfidophilus and Rps. blastica contain phosphate, their fatty acids are characterized by ester-linked, unsubstituted 3-OH-10:0 and amide-linked 3-OH-14:0 (Rb. sulfidophilus) or 3-oxo-14:0 (Rps. blastica). Lipid A of Rps. acidophila is free of phosphate and contains the rare 3-OH-16:0 fatty acid in amide linkage.The lipopolysaccharides of all three species contain 2-keto-3-deoxy-octonate (KDO) but are devoid of heptoses. Neutral sugars with the exception of glucose are lacking in the lipopolysaccharide of Rb. sulfidophilus. This shows a high galacturonic acid content. The lipopolysaccharides of Rps. acidophila and Rps. blastica have neutral sugar spectra indicative for typical O-chains (rhamnose, mannose, galactose, glucose in both species, and in Rps. blastica additionally 2-O-methyl-6-deoxy-hexose). The taxonomic value of the data is discussed.This paper is dedicated to Prof. Dr. Norbert Pfennig on the occasion of his 60th birthday     

Detection of lipoglycans in ureaplasmas.

Serotypes 3, 4, and 8 of Ureaplasma urealyticum were found to contain lipoglycans. Although the ratios of their components differed, all contained neutral sugars, fatty acids, glycerol, and phosphorus. All three became labeled when the organisms were cultivated in the presence of [14C]glucose, [14C]palmitic or [14C]oleic acids, and inorganic 32P. Only neutral sugars were found, and these consisted of mannose, glucose, and galactose. Hot phenol extracts of uninoculated and supernatant culture media contained polymeric carbohydrate, but this differed in composition from ureaplasmal lipoglycans and did not become radiolabeled. Since lipoglycans contained phosphorus but no amino sugars, they could be separated from contaminating polysaccharides by anion exchange chromatography.     

Extraction and biochemical analyses ofHelicobacter pylori lipopolysaccharides

Lipopolysaccharides were isolated from dehydratedHelicobacter pylori cells by the phenol-chloroform-petroleum ether and hot phenol/water extraction techniques. Biochemical characterization of the crude extracts indicated the following: The primary fatty acids and approximate molar ratios were 3-hydroxyoctadecanoic (2), 3-hydroxyhexadecanoic (1), and octadecanoic (1) acids. Lesser amounts of tetradecanoic, hexadecanoic, and octadecenoic acids were noted. 3-Hydroxytetradecanoic acid was not deteted in either extract. Neutral sugar analyses detected glucose, galactose, two heptose isomers, and an unidentified deoxy-hexose. Glucosamine and glucosamine phosphate were the only amino sugars found in significant quantities. Analyses of other components included ethanolamine, phosphate, and protein. 3-Deoxy-d-manno-octulosonic acid (KDO) was detected, but in lower concentrations than would be expected in comparable enterobacterial lipopolysaccharides.     

Lipopolysaccharides of Group F,a new group of vibrios

The sugar composition of the O-antigenic lipopolysaccharides isolated from Group F vibrios was analysed. 2-Keto-3-deoxy-octonate was totally absent from the lipopolysaccharides. As common component sugars, glucose, galactose, L-glycero-D-mannoheptose, and glucosamine were present. The Group F vibrios examined were found to be divided into two groups, designated tentatively as groups I and II, on the basis of the pattern of the sugar composition of their lipopolysaccharides. As additional sugar components, mannosamine, quinovosamine and two unidentified amino sugars, F1 and F2, were present in group I, while rhamnose, galactosamine, an unidentified amino sugar, F3, and a relatively high content of D-glycero-D-mannoheptose were found in group II.     

Lipopolysaccharides from Mesorhizobium huakuii and Mesorhizobium ciceri: chemical and immunological comparative data

Lipopolysaccharides of two Mesorhizobium species of different host specificity were compared: M. huakuii and M. ciceri. M. huakuii sp. was represented by five strains with special consideration of M. huakuii IFO 15243(T). SDS/PAGE profiles revealed that all M. huakuii LPS preparations contained low molecular mass fractions (LPS-II) of the same molecular size. All of lipopolysaccharides contained high molecular mass fractions (LPS-I). However, the high molecular mass fraction from each strain possessed an individual molecular size distribution pattern. The crossreactivity of blotted lipopolysaccharides with rabbit polyclonal antibodies against Mesorhizobium huakuii IFO 15243(T) whole bacteria indicated the presence of common epitope(s) within the investigated Mesorhizobium huakuii strains. Moreover, LPS from M. huakuii S52 also reacted with anti M. ciceri HAMBI 1750 serum showing that there are epitopes common for different mesorhizobial species. LPS isolated from Mesorhizobium huakuii strain IFO 15243(T) contained neutral sugars: L-6-deoxytalose, L-rhamnose, D-galactose and D-glucose, aminosugars:D-quinovosamine, D-glucosamine, D-2,3-diamino-2,3-dideoxyglucose and D-galacturonic and D-glucuronic acids. In the LPS preparation, fatty acids typical for Mesorhizobium strains were detected. 3-Hydroxydodecanoic, 3-hydroxy-iso-tridecanoic, 3-hydroxyeicosanoic, 3-hydroxyheneicosanoic and 3-hydroxydocosenoic acids were the major amide linked fatty acids, while iso -heptadecanoic, eicosanoic, docosenoic, as well as 27-hydroxyoctacosanoic and 27-oxooctacosanoic acids were the dominant ester linked fatty residues.     

Chemical composition of lipopolysaccharide from Azospirillum lipoferum

Abstract The lipopolysaccharide isolated from Azospirillum lipoferum strain SpBr17 (ATCC29709) was proven to be composed from 7 neutral sugars, two of which, rhamnose and glucose, were the major constituents. Two heptoses, l -glycero- d -mannoheptose and d -glycero- l -mannoheptose were identified. Among 8 fatty acids isolated from the lipopolysaccharide only 3-hydroxypalmitic acid was amide-bound. The approximate molar ratios of the constituents 3-deoxy- l -mannooctulosonic acid : glucosamine : amide-linked fatty acids : ester-linked fatty acids : phosphate were 0.8 : 4 : 2 : 4 : 2.5.     

Chemical Composition of Exosporium from Spores of Bacillus cereus

Homogeneous fragments of exosporium were extricated in centigram amounts from dormant spores of Bacillus cereus and analyzed for intrinsic constituents. The membrane proved to be chemically complex but not unique, consisting mainly of protein (52%), amino and neutral polysaccharides (20%), lipids (18%), and ash (4%). Seventeen common amino acids were identified by chromatography, and were present in usual proportions except for low levels of cystine-cysteine, methionine, tyrosine, and histidine. Glucosamine was the only amino sugar, and glucose and rhamnose were the principal neutral sugars. The lipid fraction contained 5.5% cardiolipin as the only phospholipid, 12.5% neutral lipids, and at least 19 fatty acids, among which normal C(16) and C(18) ones predominated. Calcium and phosphorus occurred in the ash. Small amounts of teichoic, ribonucleic, and dipicolinic acids were believed to represent contamination.     

A transfer of carbon atoms from fatty acids to sugars and amino acids in yellow lupine (Lupinus luteus L.) seedlings

The metabolism of 14C-acetate was investigated during the in vitro germination of yellow lupine seeds. Carbon atoms (14C) from the C-2 position of acetate were incorporated mainly into amino acids: aspartate, glutamate, and glutamine and into sugars: glucose, sucrose, and fructose. In contrast to this, 14C from the C-1 position of acetate was released mainly as 14CO2. Incorporation of 1-14C and 2-14C from acetate into amino acids and sugars in seedling axes was more intense when sucrose was added to the medium. However, in cotyledons where lipids are converted to carbohydrates, this process was inhibited by exogenous sucrose. Since acetate is the product of fatty acid beta-oxidation, our results indicate that, at least in lupine, seed storage lipids can be converted not only to sucrose, but mainly to amino acids. Inhibitory effects of sucrose on the incorporation of 14C from acetate into amino acids and sugars in cotyledons of lupine seedlings may be explained as the effect of regulation of the glyoxylate cycle by sugars.     

Hyphal Wall Compositions of Marine and Terrestrial Fungi of the Genus Leptosphaeria

Hyphal wall compositions of six Leptosphaeria species were compared to assess whether gross changes have occurred in the hyphal wall chemistry of closely related fungi which have become ecologically restricted to marine or terrestrial habitats. Unfractionated, lipid-extracted hyphal walls of each Leptosphaeria species had qualitatively identical compositions consisting of glucose, mannose, galactose, glucosamine, amino acids, and traces of galactosamine. Quantitative analyses showed that the hyphal wall components varied from species to species. Qualitative compositions of alkali-soluble wall fractions from each species were identical and contained the same sugars found in the unfractionated walls. The alkali-insoluble residues contained glucose, glucosamine, and amino acids. The alkali-soluble fractions were composed predominantly of glucose, galactose, and mannose. The alkali-insoluble fractions contained high concentrations of glucose and glucosamine and relatively low concentrations of amino acids.     

Studies on the influence of foliar nutrient sprays on the root exudation pattern in four crop plants

Summary Studies conducted to examine the exudation pattern of amino acids and sugars in four crop plants,viz sorghum, sunnhemp, ragi, and tomato indicated that in all, 17 known amino acids and 4 sugars were exuded and that the number and nature of the exuded amino acids and sugars differed with the plant species and with the age of plant. Glutamic and aspartic acids were found to be present in the exudates of all the plant species at all stages of plant growth examined. The quantities of amino acids and sugars differed with plant species and the maximum quantity of the chemicals was exuded during the early stages of plant growth. Glutamic acid among amino acids, and glucose among sugars, were always present in higher concentrations than the others, in the exudates in all the four crop plants.Foliar application of nitrogen in the form of NaNO₃ and phosphorus as Na₂HPO₄, was found to alter the exudation pattern of amino acids and sugars and such influence differed in different plant species. There was a general increase in the total concentration of amino acids and a decrease in sugar content in the exudates after treatment of the foliage with N, while a decrease in the amino acid content and increase in total sugars with P-treatment was observed.     

不同品种药用菊花中游离糖类及游离氨基酸含量的HPLC分析

为比较不同品种菊花的品质特征,本文采用HPLC法测定其游离糖类及游离氨基酸的含量.结果表明,不同品种菊花中游离糖类主要成分是果糖和葡萄糖,蔗糖和麦芽糖未检出,总游离糖类含量约为9.07～18.76g/1O0g,其中黄山贡菊中含量最高,为18.76g/100g,黄菊中最低,为9.07g/100g;其总游离氨基酸含量约为0.563～2.806g/100g,七月菊中含量最高,为2.806g/100g,杭白菊中最低,为0.563g/100g.     

Effects of nutrient addition on phytoplankton in Lake Nakanuma,Japan I. Changes in biochemical components

Effects of nutrient conditions on biochemical components (sugars and amino acids) of phytoplankton were examined in Lake Nakanuma in Japan. Phosphate, ammonium and silicate were added to water samples collected at 0 m, which were incubated for 15 days in situ. Chlorophyll a in phosphate-added samples increased much more than that in other samples. Total amino acids and total neutral sugars in phosphate-added samples also increased more than those in other samples. The increase of total amino acids and chlorophyll began faster than that of total neutral sugars during the first 5 days of incubation. Total neutral sugars in the phosphate-added samples increased rapidly after 8 days. The composition of amino acids did not change so much. However, the composition of neutral sugars changed according to the different nutrient addition. Phosphate-added samples changed greatly during the incubation. These changes were explained at least partly by changes in nutrient conditions. Addition of limiting nutrients decreased glucose content, whereas depletion of nutrients increased the content. This study indicates that measurements of the biochemical components contribute to the analysis of effects of nutrients on phytoplankton in natural waters.     

Hemagglutinin activity of human plasma fibronectin.

Purified human plasma fibronectin at concentrations of about 30 microgram/ml was found to agglutinate trypsin-treated erythrocytes from certain species. The hemagglutination reaction was inhibited by specific antibodies to fibronectin, by relatively low concentrations of polyamines and by higher concentrations of basic amino acids and nonacetylated amino sugars. The divalent cations Ca2+ and Mg2+ and the chelating agent ethylenediaminetetraacetate did not affect the reaction. None of the neutral amino acids, neutral sugars or polyanions tested was inhibitory. The results imply that plasma fibronectin is capable of interacting with cell surfaces and support the idea of a similarity between cellular and plasma fibronectins.     

The composition of β-lactamase I and β-lactamase II from Bacillus cereus 569/H

1. Crystalline beta-lactamase I from Bacillus cereus 569/H yielded only amino acids on acid hydrolysis, but crystalline beta-lactamase II from the same organism yielded also substantial quantities of neutral sugars and amino sugars. 2. Analysis with an amino acid analyser indicated that the two enzymes were similar though not identical in overall amino acid composition. Analysis of neutral and amino sugars as their silyl derivatives by gas-liquid chromatography showed that the carbohydrate moiety of beta-lactamase II contained residues of glucose, galactose, mannose, fucose, glucosamine and galactosamine. 3. After oxidation and hydrolysis both beta-lactamases gave small amounts of cysteic acid. After treatment of inactive Zn(2+)-free beta-lactamase II with N-ethylmaleimide or iodoacetate enzymic activity was not restored by the addition of Zn(2+).     

Chemical analyses,local Shwartzman reactivity,and body weight-decreasing activity of aqueous-phenol extracts of Mycoplasma salivarium cells

Aqueous-phenol extracts of Mycoplasma salivarium ATCC 23064 cells (APM) showed demonstrable differences from lipopolysaccharides (LPSs) of Veillonella rodentium ATCC 17743. These were as follows: smaller amounts of amino sugars and an absence of 2-keto-3-deoxyoctonate; local Shwartzman reactivity and body weight-decreasing activity, even though the activities were rather weak compared with those of LPSs. Therefore, phenol-water extractive components of Mycoplasma salivarium might be of pathogenic importance in mediating damaging effects on the periodontium.     

Crystallization and partial characterization of Staphylococcus aureus hyaluronate lyase

Staphylococcus aureus, strain 1801, hyaluronate lyase was purified and crystallized to homogeneity as ascertained by chromatography and disc-gel electrophoresis. Purification procedures included sequential ammonium sulfate fractionation, acetone precipitation, Sephadex gel filtration, and ion exchange chromatography. During its passage through the cation exchange column, the hyaluronate lyase was resolved into two minor and one major fraction. The major peak, which was found to be cationic, was further characterized and designated as Fraction III. Carbohydrate analysis showed the presence of neutral sugars galactose, glucose, and mannose in the ratio of 1:3:6. Amino sugars galactosamine and glucosamine (or mannosamine) were present in a ratio of 1:1. Quantitative amino acid analysis of the Fraction III showed a relative abundance of the basic amino acids lysine and histidine.     

Quantitative microanalysis of cell walls of Saprolegnia diclina humphrey and Tremella mesenterica fries

Cell walls of the fungi Saprolegnia diclina Humphrey and Tremella mesenterica Fries were analyzed quantitatively. Particular attention was paid to the hydrolysis and analysis of neutral sugars, amino sugars and amino acids. These components, together with total lipids, total uronic acids and the ashed residue, accounted for more than 90% by weight of the original dry cell wall preparation. There were substantial losses of amino acids during hydrolysis; however, analytical recovery approached 100% when total protein was calculated from the total nitrogen analysis. The analytical procedures were reproducible (±3% for amino acids and amino sugars, and ±5–10% for other components) when applied to individual cell wall preparations. However, even under carefully standardized conditions, different cell wall preparations from the same species showed variable composition.Glucose was the predominant neutral sugar in the cell wall polymers of both species. The amino acid compositions were remarkable in that neither species contained detectable levels of cyst(e)ine. Hydroxyproline was detected in both species. The report from Tremella mesenterica is the first for this imino acid from the cell wall of a Basidiomycete.     

Platelet plasma membrane lectin activity

The lectin activity of human platelet and erythrocyte membranes was evaluated using trypsinized, formalinized erythrocytes from eight species. Platelet membranes had the greatest lectin activity against cow erythrocytes, but also had significant activity against human, sheep, electric eel, and rabbit erythrocytes. In contrast, erythrocyte membranes only had low lectin activity against electric eel erythrocytes with no activity against the other types of erythrocytes tested. The platelet membrane lectin activity was found to reside in protein molecules on the external surface of the platelet plasma membrane. The lectin activity of platelet membranes was inhibited by amino sugars and some basic amino acids: N-acetylated amino sugars and other neutral sugars were without effect. These results demonstrate that the external surface of the platelet plasma membrane has a specific lectin activity.