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E Ito

Publications and source records attributed to E Ito.

At least 289 records · Page 16Linked to original sources

Structural studies on the acidic polysaccharide of Bacillus cereus AHU 1356 cell walls.

Structural studies were carried out on the acidic polymer fraction isolated from lysozyme digests of the N-acetylated cell walls of Bacillus cereus AHU 1356. The acidic polymer fraction contained glucosamine, galactose, rhamnose, glycerol and phosphorus in a molar ratio of 1:1:2:1:1, together with small amounts of glycopeptide components and muramic acid 6-phosphate. The hydrogen fluoride treatment led to removal of glycerol and phosphorus from the polymer without loss of other components. Results of the NaIO4 oxidation, methylation and proton magnetic resonance spectroscopy of the native and dephosphorylated preparations, in combination with data of the analysis of oligosaccharides obtained from partial hydrolysis of polysaccharide, led to the most likely structure of the repeating units of the acidic polysaccharide chain, ----4)N-acetylglucosaminyl-(alpha 1----3)rhamnosyl(alpha 1----3)galactosyl(alpha 1----4)[sn-glycerol 1-phospho-2]rhamnosyl(alpha 1----.

Bacillus cereus↗

Structural studies on the linkage unit of ribitol teichoic acid of Lactobacillus plantarum.

Structural studies were carried out on the linkage unit which joins ribitol teichoic acid to peptidoglycan in the cell walls of Lactobacillus plantarum AHU 1413. The heating of the cell walls at pH 2.5 led to release of only 5% of ribitol teichoic acid components as water-soluble material. In contrast, the same treatment of the cell walls after N-acetylation led to release of about 80% of the teichoic acid moiety, giving a teichoic-acid-linked sugar preparation which contained about equimolar amounts of mannosamine, glucosamine and glycerol as minor components. The teichoic-acid-linked sugar was hydrolyzed by mild alkaline treatment into a disaccharide, N-acetylmannosaminyl(beta 1----4)N-acetylglucosamine and ribitol teichoic acid linked to glycerol. The Smith degradation of the N-acetylated cell walls gave a characteristic fragment, 1,2-ethylenediol-phospho-glycerol-phospho-N-acetylmannosaminyl(beta 1----4) N-acetylglucosamine. Furthermore, when the intact cell walls were subjected to the NaNO2 treatment followed by NaBH4 reduction, the ribitol teichoic acid moiety was recovered for the most part in the water-soluble polymer fraction, from which a sugar, N-acetylmannosaminyl-2,5-anhydromannitol, was released by mild alkaline treatment. These results lead to the conclusion that the ribitol teichoic acid chain in the intact cell walls of this organism is linked to peptidoglycan through a unique linkage unit, glycerol-phospho-N-acetylmannosaminyl(beta 1----4)-glucosamine. The anomalous stability of the linkage between the teichoic acid moiety and peptidoglycan against acid hydrolysis seems to be accounted for by the involvement of the N-substituted glucosamine residue in the phosphodiester bridge that joins the two polymers.

Binding Sites↗

Structural studies on the linkage unit between poly(galactosylglycerol phosphate) and peptidoglycan in cell walls of Bacillus coagulans.

Structural studies were carried out on the linkage units in the teichoic-acid--glycopeptide complexes isolated from lysozyme digests of the cell walls of Bacillus coagulans AHU 1366. On treatment with 47% hydrogen fluoride, the complexes gave a disaccharide characterized as glucosyl(beta 1----4)N-acetylglucosamine together with major fragments, galactosyl(alpha 1----2)glycerol. By means of Smith degradation and partial acid hydrolysis, the teichoic acid chain was shown to be composed of the repeating units, galactosyl(alpha 1----2)glycerol-3(1)-phosphate, which were joined by phosphodiester bonds at C-6 of the galactose residues. The mild alkaline hydrolysis of teichoic-acid-linked glycan fragments yielded teichoic acid chains and disaccharide-linked glycan fragments, from which the disaccharide, glucosyl(beta 1----4)N-acetylglucosamine, was liberated by mild acid hydrolysis, whereas the same disaccharide linked to the teichoic acid chain was obtained by direct heating of the cell walls at pH 2.5. In addition, the presence of specialized glycerol phosphate units in the linkage region was shown by the isolation of tris(glycerol phosphate)3-glucosyl(beta 1----4)N-acetylglucosamine from the products of the Smith degradation of the teichoic-acid--glycopeptide complexes. Thus, it is concluded that the poly(galactosylglycerol phosphate) chain in the cell walls of B. coagulans AHU 1366 is linked to peptidoglycan through a novel linkage unit, bis(glycerol phosphate)-3-glucosyl(beta 1----4)N-acetylglucosamine.

Bacillus↗

Characterization of a novel linkage unit between ribitol teichoic acid and peptidoglycan in Listeria monocytogenes cell walls.

The structure of the linkage unit between ribitol teichoic acid and peptidoglycan in the cell walls of Listeria monocytogenes EGD was studied. A teichoic-acid--glycopeptide preparation isolated from lysozyme digests of the cell walls of this strain contained mannosamine, glycerol, glucose and muramic acid 6-phosphate in an approximate molar ratio of 1:1:2:1, together with large amounts of glucosamine and other components of teichoic acid and glycopeptides. A teichoic-acid-linked sugar preparation, obtained by heating the cell walls at pH 2.5, also contained glucosamine, mannosamine, glycerol and glucose in an approximate molar ratio of 25:1:1:2. Part of the glucosamine residues were shown to be involved in the linkage unit. Thus, on mild alkaline hydrolysis, the teichoic-acid-linked sugar preparation gave a disaccharide characterized as N-acetylmannosaminyl(beta 1----4)-N-acetylglucosamine [ManNAc(beta 1----4)GlcNAc] in addition to the ribitol teichoic acid moiety, whereas the teichoic-acid - glycopeptide was separated into disaccharide-linked glycopeptide and the ribitol teichoic acid moiety by the same procedure. Furthermore, Smith degradation of the cell walls gave a characteristic fragment, EtO2-P-Glc(beta 1----3)Glc(beta 1----1/3)Gro-P-ManNAc(beta 1----4)GlcNAc (where EtO2 = 1,2-ethylenediol and Gro = glycerol). The results lead to the conclusion that in the cell walls of this organism, the ribitol teichoic acid chain is linked to peptidoglycan through a novel linkage unit, Glc(beta 1----3)Glc(beta 1----1/3)Gro-P-(3/4)ManNAc-(beta 1----4)GlcNAc.

Cell Wall↗

A common linkage saccharide unit between teichoic acids and peptidoglycan in cell walls of Bacillus coagulans.

Teichoic acid-glycopeptide complexes were isolated from lysozyme digests of the cell walls of Bacillus coagulans AHU 1631, AHU 1634, and AHU 1638, and the structure of the teichoic acid moieties and their linkage regions was studied. On treatment with hydrogen fluoride, each of the complexes gave a hexosamine-containing disaccharide, which was identified to be glucosyl(beta 1----4)N-acetylglucosamine, in addition to dephosphorylated repeating units of the teichoic acids, namely, galactosyl(alpha 1----2)glycerol and either galactosyl(alpha 1----2)[glucosyl(alpha 1----1/3)]glycerol (AHU 1638) or galactosyl(alpha 1----2)[glucosyl(beta 1----1/3)]glycerol (AHU 1631 and AHU 1634). From the results of Smith degradation, methylation analysis, and partial acid hydrolysis, the teichoic acids from these strains seem to have the same backbone chains composed of galactosyl(alpha 1----2)glycerol phosphate units joined by phosphodiester bonds at C-6 of the galactose residues. The presence of the disaccharide, glucosyl(beta 1----4)N-acetylglucosamine, in the linkage regions between teichoic acids and peptidoglycan was confirmed by the isolation of a disaccharide-linked glycopeptide fragment from each complex after treatment with mild alkali and of a teichoic acid-linked saccharide from each cell wall preparation after treatment with mild acid. Thus, it is concluded that despite structural differences in the glycosidic branches, the teichoic acids in the cell walls of the three strains are linked to peptidoglycan through a common linkage saccharide, glucosyl (beta 1----4) N-acetylglucosamine.

Bacillus↗

Biosynthesis of UDP-N-acetyl-D-glucosaminuronic acid and UDP-N-acetyl-D-mannosaminuronic acid in Micrococcus luteus.

The occurrence and formation of UDP-N-acetyl-D-glucosaminuronic acid (UDP-GlcNAcA) and UDP-N-acetyl-D-mannosaminuronic acid (UDP-ManNAcA) were studied in Micrococcus luteus ATCC 4698. UDP-N-acetylhexosaminuronic acid separated from D-cycloserine-inhibited cells was shown to be a mixture of UDP-GlcNAcA and UDP-ManNAcA in the ratio of 87:13, whereas that obtained from untreated cells was a 96:4 mixture of these two nucleotides. Crude enzyme preparations obtained from the supernatant fraction of cells catalyzed the NAD+-dependent conversion of UDP-GlcNAc into UDP-GlcNAcA and UDP-ManNAcA. Studies on the partial separation and properties of enzymes revealed that UDP-GlcNAcA is synthesized directly from UDP-GlcNAc by the action of UDP-GlcNAc dehydrogenase and that UDP-ManNAcA is synthesized from UDP-GlcNAc through the successive actions of UDP-GlcNAc 2-epimerase and UDP-ManNAc dehydrogenase. However, enzymatic conversion of UDP-GlcNAcA to UDP-ManNAcA was not detected. Ammonium sulfate protects both dehydrogenases from inactivation during storage and incubation. Partially purified UDP-GlcNAc dehydrogenase required dithiothreitol and the particulate fraction for its full activity. The apparent Km values of UDP-GlcNAc dehydrogenase for UDP-GlcNAc and NAD+ were 0.28 and 1.43 mM, respectively. The optimum pH of this enzyme was higher than 9 in Tris-HCl buffer. p-Chloromercuribenzoate at 27 microM as well as 10 mM ethanol almost completely inhibited the UDP-GlcNAc dehydrogenase reaction.

Carbohydrate Dehydrogenases↗

Structure of the linkage units between ribitol teichoic acids and peptidoglycan.

The structure of the linkage regions between ribitol teichoic acids and peptidoglycan in the cell walls of Staphylococcus aureus H and 209P and Bacillus subtilis W23 and AHU 1390 was studied. Teichoic acid-linked saccharide preparations obtained from the cell walls by heating at pH 2.5 contained mannosamine and glycerol in small amounts. On mild alkali treatment, each teichoic acid-linked saccharide preparation was split into a disaccharide identified as N-acetylmannosaminyl beta(1----4)N-acetylglucosamine and the ribitol teichoic acid moiety that contained glycerol residues. The Smith degradation of reduced samples of the teichoic acid-linked saccharide preparations from S. aureus and B. subtilis gave fragments characterized as 1,2-ethylenediol phosphate-(glycerolphosphate)3-N-acetylmannosaminyl beta(1----4)N- -acetylxylosaminitol and 1,2-ethylenediolphosphate-(glycerol phosphate)2-N-acetylmannosaminyl beta(1----4)N-acetylxylosaminitol, respectively. The binding of the disaccharide unit to peptidoglycan was confirmed by the analysis of linkage-unit-bound glycopeptides obtained from NaIO4 oxidation of teichoic acid-glycopeptide complexes. Mild alkali treatment of the linkage-unit-bound glycopeptides yielded disaccharide-linked glycopeptides, which gave the disaccharide and phosphorylated glycopeptides on mild acid treatment. Thus, it is concluded that the ribitol teichoic acid chains in the cell walls of the strains of S. aureus and B. subtilis are linked to peptidoglycan through linkage units, (glycerol phosphate)3-N-acetylmannosaminyl beta(1----4)N-acetylglucosamine and (glycerol phosphate)2-N-acetylmannosaminyl beta(1----4)N-acetylglucosamine, respectively.

Bacillus subtilis↗

Acute effects of nifedipine on patients with chronic obstructive lung disease.

The bronchodilatory effect of nifedipine (Adalat) and the change of arterial blood gas in 5 normal and 25 patients with chronic obstructive lung disease (COLD) were investigated. 10 or 20 mg of nifedipine was orally administered. Before and 2 h after the administration, blood pressure (BP), heart rate (HR), arterial blood gas, respiratory resistance (Rrs) were measured and spirometry was assessed. Decrease of BP and increase of HR were observed in both normal and patient group. The patient group showed an increase of forced vital capacity (FVC) by 150 ml and forced expiratory volume (FEV 1.0) by 140 ml in average. In spite of the improvement of spirometry and Rrs, delta AaPO2 (alveolar arterial oxygen pressure difference) was increased in the patient group. It was considered, that the effect on the cardiovascular system was more remarkable than the bronchodilation, and the use of the drug alone as a bronchodilator may not be appropriate. It may be particularly effective in the cases with cardiovascular disease complicated with COLD.

Adult↗

Biosynthesis of N-acetylmannosaminuronic-acid-containing cell-wall polysaccharide of Bacillus subtilis.

The particulate enzyme from Bacillus subtilis AHU 1031 catalyzed the synthesis of a polysaccharide and glycolipids from UDP-N-acetylmannosaminuronic acid (UDP-ManNAcUA), UDP-N-acetylglucosamine (UDP-GlcNAc), and UDP-glucose (UDP-Glc). The polysaccharide synthesis required UDP-ManNAcUA and UDP-GlcNAc, proceeded optimally at pH 8.5 and in the presence of 5 mM MgCl2 and 2.5 mM dithiothreitol, and was stimulated by the addition of UDP-Glc. The molar ratio of ManNAcUA, GlcNAc, and Glc incorporated into polysaccharide was calculated to be 1:1:1.8 from chemical analysis involving reduction with water soluble carbodiimide; its relative molecular mass was estimated to be 12000. The analysis of Smith degradation products revealed that the polysaccharide backbone is composed of repeating trisaccharide units comprising ManNAcUA, GlcNAc, and Glc. Based on the data regarding the time course of the incorporation of glucose into the polysaccharide, extra glucose seems to be attached to the polysaccharide backbone as lateral branches. The saccharide moieties of the glycolipids were identified as GlcNAc, ManNAcUA-GlcNAc, and Glc-ManNAcUA-GlcNAc from several analytical criteria. The addition of antibiotic 24010, a tunicamycin-like antibiotic, at 10 micrograms/ml resulted in almost complete inhibition of the synthesis of glycolipids and polysaccharide. It is therefore concluded that the glycolipids function as intermediates in polysaccharide formation. Incubation of the ManNAcUA-GlcNAc-linked lipid. (labeled in the ManNAcUA moiety) with the particulate enzyme and UDP-Glc resulted incorporation of radioactivity into a trisaccharide-linked lipid and a polysaccharide. These results suggest that the particulate enzyme utilizes the trisaccharide moiety of the Glc-ManNAcUA-GlcNAc-linked lipid for the elongation of the main polysaccharide chain presumed to be cell wall acidic polysaccharide of this strain.

Anti-Bacterial Agents↗

The primary structure of teichuronic acid in Bacillus subtilis AHU 1031.

Structural studies were carried out on the acidic polysaccharide fraction obtained from lysozyme digest of the cell walls of Bacillus subtilis AHU 1031. The polysaccharide fraction contained N- acetylmannosaminuronic acid ( ManNAcA ), N-acetylglucosamine (GlcNAc), glucose, glycerol and phosphorus in a molar ratio of 2:2:4:1:1, together with glycopeptide components. The results of analyses involving Smith degradation, chromium trioxide oxidation, methylation and proton magnetic resonance spectroscopy led to the conclusion that the backbone chain of the polysaccharide has the repeating unit----6)Glc(alpha 1----3/4) ManNAcA (beta 1----4)GlcNAc(beta 1----. About 50% of the N-acetylglucosamine residues in the backbone chain seem to be substituted at C-3 by the glycosidic branches, glycerol phospho-6-glucose, while the other half seem to be substituted by glucose.

Bacillus subtilis↗

Liver injuries induced by cyclochlorotine isolated from Penicillium islandicum.

Sequential morphological changes in murine liver induced by cyclochlorotine (CC), a secondary metabolite of Penicillium islandicum were investigated by transmission electron microscopy and scanning electron microscopy. Within 15 min after the administration of CC there was a marked dilatation of Disse's space around the portal triads, and the exudates then poured out into the space which was formed by the invagination of the hepatocyte plasma membrane. Shortly after the invagination was completed, the connection between Disse's space and the invaginated space was pinched off, so that this space became a membrane-bound vacuole. After dehydration, the vacuoles became granular. The liver injuries induced by CC were influenced by various pretreatments. The results indicate that drug-metabolizing systems mediated by cytochrome p-450 in the hepatocytes may play an important role in the hepatotoxicity of CC.

Aging↗

Quantification of myonecrosis and comparison of necrotic activity of snake venoms by determination of creatine phosphokinase activity in mice sera.

Serum creatine phosphokinase (CK) values were measured in venom-injected mice. The increase of CK levels was proportional to both the amount of venom injected and the myonecrotic actions determined microscopically. The CK isoenzymes were regarded as due to the predominant increase of CK-MM, which was considered to be of skeletal muscle origin in the venom-injected area. No CK isozyme variants were found by electrophoretic examination. For a comparative study on necrotic intensity, nine snake venoms were examined. The elapid venoms induced higher values of serum CK with a stronger necrotic effect than the crotalid venoms, and the data agreed with the histological findings. On application of the antivenom, an initially elevated CK value was significantly decreased and no severe damage was observed under light microscopy. Since the changes of serum CK activities paralleled necrotic intensity, the CK value is useful as an indicator of myonecrosis due to snake venom.

Animals↗

N-acetylmannosaminyl(1----4)N-acetylglucosamine, a linkage unit between glycerol teichoic acid and peptidoglycan in cell walls of several Bacillus strains.

The structure of teichoic acid-glycopeptide complexes isolated from lysozyme digests of cell walls of Bacillus subtilis (four strains) and Bacillus licheniformis (one strain) was studied to obtain information on the structural relationship between glycerol teichoic acids and their linkage saccharides. Each preparation of the complexes contained equimolar amounts of muramic acid 6-phosphate and mannosamine in addition to glycopeptide components and glycerol teichoic acid components characteristic of the strain. Upon treatment with 47% hydrogen fluoride, these preparations gave, in common, a hexosamine-containing disaccharide, which was identified as N- acetylmannosaminyl (1----4) N-acetylglucosamine, along with large amounts of glycosylglycerols presumed to be the dephosphorylated repeating units of teichoic acid chains. The glycosylglycerol obtained from each bacterial strain was identified as follows: B. subtilis AHU 1392, glucosyl alpha (1----2)glycerol; B. subtilis AHU 1235, glucosyl beta(1----2) glycerol; B. subtilis AHU 1035 and AHU 1037, glucosyl alpha (1----6)galactosyl alpha (1----1 or 3)glycerol; B. licheniformis AHU 1371, galactosyl alpha (1----2)glycerol. By means of Smith degradation, the galactose residues in the teichoic acid-glycopeptide complexes from B. subtilis AHU 1035 and AHU 1037 and B. licheniformis AHU 1371 were shown to be involved in the backbone chains of the teichoic acid moieties. Thus, the glycerol teichoic acids in the cell walls of five bacterial strains seem to be joined to peptidoglycan through a common linkage disaccharide, N- acetylmannosaminyl (1----4)N-acetylglucosamine, irrespective of the structural diversity in the glycosidic branches and backbone chains.

Acetylglucosamine↗