COMPOSITION OF PURIFIED MUCOPEPTIDE FROM THE WALL OF AEROBACTER CLOACAE

1962 ◽  
Vol 8 (1) ◽  
pp. 89-98 ◽  
Author(s):  
A. J. Schocher ◽  
S. T. Bayley ◽  
R. W. Watson
Keyword(s):  
Glutamic Acid ◽  
Acid Hydrolysis ◽  
Teichoic Acid ◽  
Sodium Lauryl Sulphate ◽  
Diaminopimelic Acid ◽  
Peptide Chain ◽  
Muramic Acid ◽  
Aqueous Sodium ◽  
Molar Ratios ◽  
Saturated Aqueous Sodium

Molecular proportions of glucosamine, muramic acid, alanine, glutamic acid, and diaminopimelic acid in purified mucopeptide (MP) from the wall of Aerobacter cloacae NRC 492 have been estimated. Crude walls, obtained by fractional centrifugation of disintegrated log-phase cells, were treated with trypsin, ribonuclease, and pepsin to remove cytoplasmic contaminants. Of a number of protein solvents tested on the isolated walls, saturated aqueous sodium lauryl sulphate gave the purest MP in the highest yield. Molar ratios of glucosamine, muramic acid, alanine, glutamic acid, and diaminopimelic acid in hydrolyzates of the MP, corrected for the loss of hexosamines under the conditions of acid hydrolysis, were 2:2:3:2:2 respectively. These ratios indicate the presence of two types of peptide, with one peptide chain attached to each disaccharide unit. Analysis showed the absence of phosphate and therefore of teichoic acid from the purified MP.

scholarly journals Autolysis of cell walls of Bacillus stearothermophilus B65 and the chemical structure of the peptidoglycan

1970 ◽  
Vol 118 (5) ◽  
pp. 859-868 ◽  
Author(s):  
W. D. Grant ◽  
A. J. Wicken
Keyword(s):  
Glutamic Acid ◽  
Cell Walls ◽  
Bacillus Stearothermophilus ◽  
Teichoic Acid ◽  
Paper Electrophoresis ◽  
Diaminopimelic Acid ◽  
Peptide Fragments ◽  
Acid Complex ◽  
Acid Hydrolysate ◽  
Peptide Mixture

1. The cell walls of Bacillus stearothermophilus B65 contain glucosamine, muramic acid, alanine, α∈-diaminopimelic acid (Dap), glutamic acid, aspartic acid, glycine, and serine in the molecular proportions 0.60:0.64:2.30:0.85:1.00:0.11:0.13:0.31. 2. Both d- and l-alanine are present, but glutamic acid and diaminopimelic acid are present only as the d- and meso-isomers respectively. 3. The peptide fragments Ala-Dap, Dap-Ala, and Dap-Ala-Dap have been isolated from a partial acid hydrolysate of the cell walls. 4. The major products of autolysis of the cell wall were d-alanine, a peptide mixture, peptidoglycan material and a peptidoglycan–teichoic acid complex. 5. Separation of the peptide mixture into ten major peptides was achieved by DEAE-Sephadex and paper chromatography, and paper electrophoresis. 6. The structures of these peptides have been determined and they fall into four groups, the individual members of each group differing only in number or position of carboxamide substituents. 7. The structures are I, a tripeptide l-Ala–d-Glu-meso-Dap; II, a pentapeptide made up by the tripeptide (I) linked through the ∈-amino group of its diaminopimelic acid residue to the carboxyterminal of the dipeptide meso-Dap-d-Ala; III, a heptapeptide made up by a similar linkage between the tripeptide (I) and the tetrapeptide l-Ala-d-Glu-meso-Dap-d-Ala; IV, a possible undecapeptide made up by a further tetrapeptide similarly linked to the heptapeptide (III) structure. 8. The structure of the peptidoglycan and the actions of the autolytic enzymes are discussed in terms of these peptide structures.

scholarly journals STUDIES ON THE CHEMISTRY AND IMMUNOCHEMISTRY OF CELL WALLS OF STAPHYLOCOCCUS AUREUS

1962 ◽  
Vol 116 (2) ◽  
pp. 229-245 ◽  
Author(s):  
Stephen I. Morse
Keyword(s):  
Staphylococcus Aureus ◽  
Glutamic Acid ◽  
Cell Walls ◽  
Antigenic Determinant ◽  
Intact Cell ◽  
Teichoic Acid ◽  
The Other ◽  
Aureus Strain ◽  
Muramic Acid ◽  
Sheep Erythrocytes

The cell walls of an 80/81 strain of Staphylococcus aureus (NYH-6) contain alanine, glycine, glutamic acid, lysine, muramic acid, glucosamine, and ribitol phosphate. 94 per cent of the phosphorus and 41 per cent of the glucosamine are removed by extraction of the cell walls with hot 5 per cent TCA, but significant amounts of the other constituents are not extracted by this procedure. The residue after hot TCA extraction (mucopeptide) is susceptible to lysozyme whereas the intact cell walls are resistant. Staphylococcus aureus cell walls are agglutinated by S. aureus antisera. Agglutination of the cell walls of one S. aureus strain is inhibited by absorption of antisera with cell walls of other S. aureus strains but not by absorption with S. albus cell walls. The ribitol teichoic acid can be isolated from cold TCA extracts of the cell walls. This compound consists almost entirely of ribitol phosphate and glucosamine. The isolated teichoic acid of strain NYH-6 is readily fixed to tanned sheep erythrocytes and these sensitized cells are agglutinated by S. aureus antisera. Cold TCA extracts of cell walls of other strains of S. aureus inhibit hemagglutination whereas extracts of S. albus walls do not. Studies on the inhibition of both hemagglutination and precipitation indicate that the antigenic determinant of S. aureus NYH-6 teichoic acid is ß-N-acetylglucosamine.

scholarly journals LOCATION AND COMPOSITION OF SPORE MUCOPEPTIDE IN BACILLUS SPECIES

1963 ◽  
Vol 16 (3) ◽  
pp. 593-609 ◽  
Author(s):  
A. D. Warth ◽  
D. F. Ohye ◽  
W. G. Murrell
Keyword(s):  
Heat Resistance ◽  
Glutamic Acid ◽  
Electron Micrographs ◽  
Bacillus Coagulans ◽  
Diaminopimelic Acid ◽  
Bacillus Species ◽  
Muramic Acid ◽  
Cortical Structure

Spore integuments of Bacillus coagulans were prepared containing nearly all the hexosamine and α, ϵ-diaminopimelic acid (DAP) present in intact spores. Subsequent autolytic action resulted in the destruction and removal of the residual cortical structure and "cortical membrane" leaving the appearance of the inner and outer spore coats unchanged in electron micrographs. Concurrently, all the hexosamine and DAP in the preparation was released mainly as non-diffusible mucopeptide containing alanine, glutamic acid, DAP, and all the glucosamine and muramic acid. Some diffusible peptides containing alanine, glutamic acid, and DAP were also present but there was little protein or carbohydrate. Lysozyme digestion of integument preparations from heated spores of Bacillus 636, B. subtilis, B. coagulans, and B. stearothermophilus specifically removed the residual cortex and cortical membrane with the release of the mucopeptide. In B. cereus T, only the residual cortex and part of the mucopeptide were solubilized by lysozyme. The effect of several reagents and enzymes upon the appearance and removal of hexosamine from B. coagul ans spore integuments is reported. The results show that spore mucopeptide is mainly located in the residual cortex and cortical membrane and suggest that these structures consist essentially of mucopeptide. The implications of these results in relation to the "contractile cortex" theory of heat resistance in spores are discussed.

Die Aminosäuresequenz des Ornithin und Lysin enthaltenden Mureins einiger Stämme von Lactobacillus bifidus aus dem Pansen /The amino acid sequence of the ornithine and lysine containing mureins of some strains of Lactobacillus bifidus isolated from rumen

1969 ◽  
Vol 24 (12) ◽  
pp. 1524-1528 ◽  
Author(s):  
Wilhelm Holzapfel ◽  
Vittoria Scardovi ◽  
Otto Kandler
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Glutamic Acid ◽  
Cell Walls ◽  
Teichoic Acid ◽  
The Other ◽  
Molar Ratio ◽  
Partial Hydrolysis ◽  
Muramic Acid ◽  
Diamino Acid

Cell walls of six strains of Lactobacillus bifidus (recently classified as Bifidobacterium globosum) isolated from the rumen of sheep were isolated by lysing the cells with glass beads followed by tryptic digestion. No teichoic acid could be found. The polysaccharide consists of galactose, rhamnose and glucosamine. The murein (peptidoglycan) contains MurNAc, GlcNH2NAc, Glu, Ala, and diamino acids in a molar ratio of 1:1:1:5:1. Both diamino acids, lysine and ornithine are present. In two strains they occur in about equal amounts, while in the other four strains ornithine is predominant. The lysis of the cell walls by lysozyme resulted in a mixture of two types of muropeptides: One contains lysine, the other ornithine.The amino acid sequence was determined by analysing the oligopeptides arising during acid partial hydrolysis. It was shown, that the tetrapeptides attached to the muramic acid are equal to those of other mureins: L-Ala-γ-D-Glu-L-Lys (L-Orn) -D-Ala. Glutamic acid is probably amidated, since the total hydrolysate contained slightly more than one mole of NH3 per mole of glutamic acid. The cross-liking peptide is a tri-alanine, which is bound to the ω-aminogroup of the diamino acid of one tetrapeptid and to the C-terminal D-alanine of another. Since about 2 —4% of the alanine is N-terminal in the cell wall, 10 to 20% of the interpeptide bridges are probably not cross linked. In addition 2 — 3% of the diamino acids are ω-N-terminal and therefore not substituted by a trialanine. A small percentage of D-alanine and of the diamino acids is C-terminal. The latter indicates, that some of the peptide subunits are incomplete i.e.. the terminal D-alanine is missing.

scholarly journals Die Aminosäuresequenz des Serin und Asparaginsäure enthaltenden Mureins von Bifidobacterium bifidum Orla Jensen/ The amino acid sequence of the serin and aspartic acid containing mureins of Bifidobacterium bifidum ORLA JENSEN

1970 ◽  
Vol 25 (11) ◽  
pp. 1294-1301 ◽  
Author(s):  
Dieter Koch ◽  
Karl Heinz Schleifer ◽  
Otto Kandler
Keyword(s):  
Cell Wall ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Glutamic Acid ◽  
Aspartic Acid ◽  
Cell Walls ◽  
Teichoic Acid ◽  
Bifidobacterium Bifidum ◽  
Muramic Acid ◽  
Partial Acid Hydrolysis

Cell walls of Bifidobacterium bifidum var. pennsylvanicus were isolated. The polysaccharide consisted of glucose, galactose and rhamnose. Teichoic acid was not present. The murein (peptidoglycan) contained MurNAc, GlcNH2NAc, ᴅ-Glu, Ala. ʟ-Ser, ᴅ-Asp and L-Orn in a ratio of about 1 : 1 : 1 : 2 : 1 : 1 : 1. In one batch a high amount of ʟ-glutamic acid was found. It was not a constituent of the murein since it remained in the lysozyme insensitive residue.The amino acid sequence of the murein was determined by analyzing the oligopeptides arising during partial acid hydrolysis. It was shown that the peptide subunits attached to the muramic acid are the same as in many other mureins: ʟ-Ala-ᴅ-Glu-ʟ-Orn-D-Ala. The interpeptide bridge consisted of β-ᴅ-aspartyl-ʟ-serine. Since about 35% of aspartic acid and 6% of ornithine are N-terminal in the cell wall, it was assumed that only 60% of the peptide subunits are cross-linked. 4 other strains of B. bifidum proved to contain the same type of murein. While all other strains of other species of Bifidobacterium investigated contained other types of murein, it seems likely that the Orn-Ser-Asp type of the murein is typical of B. bifidum.

Partial Lack of N-Acetyl Substitution of Glucosamine in the Peptidoglycan of the Budding Phototrophic Rhodomicrobium vannielii

1987 ◽  
Vol 42 (11-12) ◽  
pp. 1165-1170 ◽  
Author(s):  
Uwe J. Jürgens ◽  
Baldur Rieth ◽  
Jürgen Weckesser ◽  
Crawford S. Dow ◽  
Wilfried A. König
Keyword(s):  
Fatty Acids ◽  
Content Analysis ◽  
Type Structure ◽  
Diaminopimelic Acid ◽  
Strain Atcc ◽  
Muramic Acid ◽  
Cross Linkage ◽  
Rigid Layer ◽  
Rhodomicrobium Vannielii

The rigid layer and peptidoglycan fractions from two strains (ATCC 17100 and Rm 5) of the budding phototrophic Rhodomicrobium vannielii were isolated. Rigid layers of both strains contain protein in addition to peptidoglycan. They were free of polysaccharides and fatty acids. The respective peptidoglycan fractions contain glucosamine, muramic acid, ʟ-and ᴅ-alanine, ᴅ-glutamic and meso-diaminopimelic acid in approximately equimolar ratios except for a signifi­ cant lower relative ᴅ-alanine content. Analysis of partial acid hydrolysates revealed A 1 γ-type structure of Rhodomicrobium vannielii peptidoglycan (shown with strain ATCC 17100). An about 10-30% lack of N-acetylation of glucosamine was indicated. The degree of cross-linkage was found to be about 60% . No differences in peptidoglycan composition and degree of cross-linkage were found between swarmer-and chain-cells as examined with strain Rm 5.

scholarly journals The extraction of cell walls of Pseudomonas aeruginosa with aqueous phenol. The insoluble residue and material from the aqueous layers

1967 ◽  
Vol 105 (2) ◽  
pp. 759-765 ◽  
Author(s):  
K. Clarke ◽  
G. W. Gray ◽  
D. A. Reaveley
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Wall ◽  
Cell Walls ◽  
Lipid A ◽  
Diaminopimelic Acid ◽  
Insoluble Residue ◽  
Muramic Acid ◽  
Gram Negative ◽  
Gram Negative Organisms ◽  
Residue Fraction

1. The insoluble residue and material present in the aqueous layers resulting from treatment of cell walls of Pseudomonas aeruginosa with aqueous phenol were examined. 2. The products (fractions AqI and AqII) isolated from the aqueous layers from the first and second extractions respectively account for approx. 25% and 12% of the cell wall and consist of both lipopolysaccharide and muropeptide. 3. The lipid part of the lipopolysaccharide is qualitatively similar to the corresponding material (lipid A) from other Gram-negative organisms, as is the polysaccharide part. 4. The insoluble residue (fraction R) contains sacculi, which also occur in fraction AqII. On hydrolysis, the sacculi yield glucosamine, muramic acid, alanine, glutamic acid and 2,6-diaminopimelic acid, together with small amounts of lysine, and they are therefore similar to the murein sacculi of other Gram-negative organisms. Fraction R also contains substantial amounts of protein, which differs from that obtained from the phenol layer. 5. The possible association or aggregation of lipopolysaccharide, murein and murein sacculi is discussed.

scholarly journals Tracer studies on the biosynthesis of amino acids from lactate by Peptostreptococcus elsdenii

1967 ◽  
Vol 105 (1) ◽  
pp. 299-310 ◽  
Author(s):  
H. J. Somerville ◽  
J. L. Peel
Keyword(s):  
Amino Acids ◽  
Glutamic Acid ◽  
Aspartic Acid ◽  
Tricarboxylic Acid Cycle ◽  
Carbon Chain ◽  
Tricarboxylic Acid ◽  
Diaminopimelic Acid ◽  
Reaction Sequence ◽  
Tracer Studies ◽  
Cell Fractions

Peptostreptococcus elsdenii, a strict anaerobe from the rumen, was grown on a medium containing yeast extract and [1−14C]- or [2−14C]-lactate. Radioisotope from lactate was found in all cell fractions, but mainly in the protein. The label in the protein fraction was largely confined to a few amino acids: alanine, serine, aspartic acid, glutamic acid and diaminopimelic acid. The alanine, serine, aspartic acid and glutamic acid were separated, purified and degraded to establish the distribution of 14C from lactate within the amino acid molecules. The labelling patterns in alanine and serine suggested their formation from lactate without cleavage of the carbon chain. The pattern in aspartic acid suggested formation by condensation of a C3 unit derived directly from lactate with a C1 unit, probably carbon dioxide. The distribution in glutamic acid was consistent with two possible pathways of formation: (a) by the reactions of the tricarboxylic acid cycle leading from oxaloacetate to 2-oxoglutarate, followed by transamination; (b) by a pathway involving the reaction sequence 2 acetyl-CoA→crotonyl-CoA→glutaconate→glutamate.

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