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

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.

1966 ◽  
Vol 12 (5) ◽  
pp. 985-994 ◽  
Author(s):  
Peter J. DeJong ◽  
Elizabeth McCoy

Vegetative cell walls and spore walls of seven Streptomyces species representing four types of spore morphology were qualitatively analysed for their components. Amino acid and carbohydrate components (glucose, glucosamine, muramic acid, diaminopimelic acid, glutamic acid, glycine, alanine, arginine, threonine, valine, leucine, and aspartic acid) in both types of walls were identical in all species. Aspartic acid was a major component in spore walls, but a minor component in vegetative cell walls. Although organic phosphate was present in both vegetative- and spore-wall hydrolysates, the other components of teichoic acid were not found nor was teichoic acid extracted from the isolated walls by cold trichloroacetic acid. A portion of the vegetative cell wall was rendered soluble with lysozyme and separated by paper electrophoresis into two fractions detected with ninhydrin. The lysozyme-resistant portion of the vegetative cell wall showed the same major and minor components as the spore walls, which are also lysozyme resistant.


2000 ◽  
Vol 182 (4) ◽  
pp. 1154-1157 ◽  
Author(s):  
Takashi Hirao ◽  
Masahide Sato ◽  
Akira Shirahata ◽  
Yoshiyuki Kamio

ABSTRACT Spermidine and cadaverine were found to be constituents of the cell wall peptidoglycan of Anaerovibrio lipolytica, a strictly anaerobic bacterium. The peptidoglycan was degraded with theN-acetylmuramyl-l-alanine amidase and endopeptidase into two peptide fragments, peptide I and peptide II, at a molar ratio of 4:1. Peptides I and II were identified asl-alanine–d-glutamic acid(αcadaverine)γ meso-diaminopimelic acid (DAP)–d-alanine andl-alanine–d-glutamic acid(αspermidine)γ meso-DAP–d-alanine, respectively. The N1-amino group of spermidine was linked to the α-carboxyl group of the d-glutamic acid residue of peptide II.


1962 ◽  
Vol 8 (1) ◽  
pp. 89-98 ◽  
Author(s):  
A. J. Schocher ◽  
S. T. Bayley ◽  
R. W. Watson

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.


1962 ◽  
Vol 116 (2) ◽  
pp. 229-245 ◽  
Author(s):  
Stephen I. Morse

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.


1969 ◽  
Vol 113 (1) ◽  
pp. 191-193 ◽  
Author(s):  
J. B. Adams ◽  
A. R. Archibald ◽  
J Baddiley ◽  
Hilary E. Coapes ◽  
A L Davison

Cell walls of strains of Lactobacillus plantarum lacking the group D precipitinogen (a glucosylribitol teichoic acid) contain glucosylglycerol teichoic acid in which the glycosidic substituents are attached to the primary hydroxyl group of glycerol. Three distinct repeating units have been isolated from the teichoic acid preparation of strain C106, indicating either that the polymer is complex or that the wall contains a mixture of teichoic acids. Walls of streptobacteria differ from those of L. plantarum and contain neither teichoic acid nor diaminopimelic acid.


1973 ◽  
Vol 19 (8) ◽  
pp. 1049-1051
Author(s):  
Siegfried Maier

The suitability of tritiated 2,6-diaminopimelic acid (3H-DAP) as a label specific for cell walls was explored in Bacillus subtilis BC 102 grown in a medium enriched with 3H-DAP and an excess of L-lysine. Fractionation of labeled cells showed 57% of the activity in the cell wall and 28% in the membrane. Chromatography of labeled wall hydrolysates revealed two activity peaks: 62% in DAP and 29% in glutamic acid – alanine. Labeled membrane was devoid of activity in the DAP position. Chromatographic purification of the 3H-DAP improved specificity, giving 7% of the activity in the membrane and 85% in the wall. In such walls DAP accounted for 82% of the total wall activity. Therefore, only 69% of the total fixed purified 3H label remained with DAP in the wall.


1969 ◽  
Vol 24 (12) ◽  
pp. 1524-1528 ◽  
Author(s):  
Wilhelm Holzapfel ◽  
Vittoria Scardovi ◽  
Otto Kandler

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.


1970 ◽  
Vol 25 (11) ◽  
pp. 1294-1301 ◽  
Author(s):  
Dieter Koch ◽  
Karl Heinz Schleifer ◽  
Otto Kandler

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.


1971 ◽  
Vol 20 (3) ◽  
pp. 442-450 ◽  
Author(s):  
Jean Heijenoort ◽  
Daniele Menjon ◽  
Bernard Flouret ◽  
Jekisiel Szulmajster ◽  
Jean Laporte ◽  
...  

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