Action of ethylenediaminetetraacetic acid, tris(hydroxymethyl)aminomethane, and lysozyme on cell walls of Pseudomonas aeruginosa

1968 ◽  
Vol 14 (8) ◽  
pp. 913-922 ◽  
Author(s):  
S. T. Cox Jr. ◽  
R. G. Eagon
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Wall ◽  
Cell Walls ◽  
Ethylenediaminetetraacetic Acid ◽  
Individual Cell ◽  
Intact Cell ◽  
Divalent Cations ◽  
Total Cell ◽  
The Individual ◽  
Phosphate Groups

Incubation of isolated cell walls of Pseudomonas aeruginosa with tris(hydroxymethyl)aminomethane (Tris), ethylenediaminetetraacetic acid (EDTA), and lysozyme, either singly or in combinations, results in partial solubilization of the cell walls.When cell walls were incubated with a combination of Tris and EDTA, approximately 30% of the total cell wall and 32% of the lipopolysaccharide fraction were solubilized. The following percentages of the individual cell wall constituents were solubilized: protein, 19%; carbohydrate, 35%; lipid, 5%; ash, 54%; and, phosphorus 23%. When lysozyme was included with Tris and EDTA, approximately 36% of the total cell wall and 86% of the lipopolysaccharide fraction were solubilized. Solubilization of the individual cell wall constituents was similarly increased.Phospholipids, which make up 7–8% of the intact cell wall of P. aeruginosa, were not released by incubation of the cell walls with these agents. Fatty acids in addition to those found in the lipopolysaccharide, however, were detected in the solubilized cell wall materials.Incubation systems composed of Tris, of EDTA, and of Tris and lysozyme solubilized cell walls to a lesser extent than incubation systems of Tris and EDTA, and of Tris, EDTA, and lysozyme. Incubation of cell walls in water, furthermore, was effective in solubilizing approximately 8% of the cell walls. Muramic acid containing materials were solubilized only in the presence of lysozyme.Materials liberated from cell walls which were incubated in systems containing EDTA exhibited single symmetrical Schlieren peaks characterized by uncorrected sedimentation constants of approximately 7 S, suggesting the release of homogenous subunits from the cell walls.Intra- and inter-molecular cross-linking by divalent cations via phosphate groups contained in lipoprotein and lipopolysaccharide components of the cell wall of P. aeruginosa is proposed.

scholarly journals Even Visually Intact Cell Walls in Waterlogged Archaeological Wood Are Chemically Deteriorated and Mechanically Fragile: A Case of a 170 Year-Old Shipwreck

Molecules ◽  
2020 ◽  
Vol 25 (5) ◽  
pp. 1113 ◽  
Author(s):  
Liuyang Han ◽  
Xingling Tian ◽  
Tobias Keplinger ◽  
Haibin Zhou ◽  
Ren Li ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Intact Cell ◽  
Nitrogen Adsorption ◽  
Wet Chemistry ◽  
Archaeological Wood ◽  
Waterlogged Archaeological Wood ◽  
X Ray Scattering ◽  
Crystallite Structure ◽  
Cell Wall Mechanics

Structural and chemical deterioration and its impact on cell wall mechanics were investigated for visually intact cell walls (VICWs) in waterlogged archaeological wood (WAW). Cell wall mechanical properties were examined by nanoindentation without prior embedding. WAW showed more than 25% decrease of both hardness and elastic modulus. Changes of cell wall composition, cellulose crystallite structure and porosity were investigated by ATR-FTIR imaging, Raman imaging, wet chemistry, 13C-solid state NMR, pyrolysis-GC/MS, wide angle X-ray scattering, and N2 nitrogen adsorption. VICWs in WAW possessed a cleavage of carboxyl in side chains of xylan, a serious loss of polysaccharides, and a partial breakage of β-O-4 interlinks in lignin. This was accompanied by a higher amount of mesopores in cell walls. Even VICWs in WAW were severely deteriorated at the nanoscale with impact on mechanics, which has strong implications for the conservation of archaeological shipwrecks.

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.

LYSOZYME SENSITIVITY OF THE CELL WALL OF PSEUDO-MONAS AERUGINOSA: FURTHER EVIDENCE FOR THE ROLE OF THE NON-PEPTIDOGLYCAN COMPONENTS IN CELL WALL RIGIDITY

1966 ◽  
Vol 12 (1) ◽  
pp. 105-108 ◽  
Author(s):  
K. Jane Carson ◽  
R. G. Eagon
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Cell Wall ◽  
Electron Micrographs ◽  
Cell Walls ◽  
Gram Negative Bacteria ◽  
Normal Cells ◽  
Thin Sections ◽  
Gram Negative ◽  
Cell Wall Rigidity

Electron micrographs of thin sections of normal cells of Pseudomonas aeruginosa showed the cell walls to be convoluted and to be composed of two distinct layers. Electron micrographs of thin sections of lysozyme-treated cells of P. aeruginosa showed (a) that the cell walls lost much of their convoluted nature; (b) that the layers of the cell walls became diffuse and less distinct; and (c) that the cell walls became separated from the protoplasts over extensive cellular areas. These results suggest that the peptidoglycan component of the unaltered cell walls of P. aeruginosa is sensitive to lysozyme. Furthermore, it appears that the peptidoglycan component is not solely responsible for the rigidity of the cell walls of Gram-negative bacteria.

Characterization of a protein-lipopolysaccharide complex released from cell walls of Pseudomonas aeruginosa by ethylenediaminetetraacetic acid

1969 ◽  
Vol 15 (7) ◽  
pp. 743-748 ◽  
Author(s):  
S. W. Rogers ◽  
H. E. Gilleland Jr. ◽  
R. G. Eagon
Keyword(s):  
Electron Microscopy ◽  
Pseudomonas Aeruginosa ◽  
Cell Walls ◽  
Ethylenediaminetetraacetic Acid ◽  
Gradient Centrifugation ◽  
Hollow Spheres ◽  
Gel Filtration ◽  
A Chain ◽  
Ultracentrifugal Analysis

Results from analytical ultracentrifugal analysis, Sephadex gel filtration, isopycnic density-gradient centrifugation, and polyacrylamide disc-gel electrophoresis revealed that ethylenediaminetetraacetic acid liberated a protein–lipopolysaccharide complex from cell walls of Pseudomonas aeruginosa with an estimated molecular weight of not less than 160 000 and probably about one million. Electron microscopy of this complex revealed spherules and rodlets. The diameter of the former was approximately 70 ± 10 Å while the dimensions of the latter were 70 ± 10 Å × 200 ± 50 Å. The rodlets appeared to be composed of three or more spherules arranged in a chain-like fashion. Electron microscopy of protein-free lipopolysaccharide revealed predominantly hollow spheres from 300 Å to 1500 Å in diameter, morphologically resembling membrane sacculi. It is proposed that the protein–lipopolysaccharide complex, but not the protein-free lipopolysaccharide, is representative of the in situ form of native endotoxin.

Ion behaviour in plant cell walls. IV. Selective cation binding by Sphagnum russowii cell walls

1990 ◽  
Vol 68 (4) ◽  
pp. 773-781 ◽  
Author(s):  
Conrad Richter ◽  
Jack Dainty
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Divalent Cations ◽  
Selectivity Coefficient ◽  
Weak Acid ◽  
Small Population ◽  
Cation Binding ◽  
Trivalent Cations ◽  
Using Data ◽  
A Site

Selective cation binding by Sphagnum russowii cell walls was investigated using data from bicationic potentiometric titrations of isolated cell walls. Selectivity measurements were interpreted in the context of Manning condensation. In titrations with cations of different valency, selectivity favoured the higher valency cation, as expected in Manning condensation. This selectivity generally increased with bathing pH until the wall-bound polyuronic acids became fully ionized (pH > 5). The selectivity coefficient order at full ionization was Na+–Ca2+ > Na+–La3+ > Ca2+–La3+, as predicted by the weak acid Donnan–Manning (WADM) model. Other phenomena also appear to influence binding selectivity. A small population of isolated binding sites are more effectively neutralized by univalent (or divalent) than divalent (or trivalent) cations. High selectivity for cations of lower valency at low pH also suggests a site isolation effect. In bicationic titrations involving divalent cations only, Sr24+ and Ca2+ were bound by the cell wall with approximately equal effectiveness, as predicted by the WADM model. However, cation or binding site specificities probably account for the favoured binding of Ca2+ over Mg2+ by the cell wall. Key words: ion exchange, cell wall, selectivity.

Synthesis of cell wall glucan, chitin, and protein by regenerating protoplasts and mycelia of Trichoderma reesei

1990 ◽  
Vol 36 (3) ◽  
pp. 211-217 ◽  
Author(s):  
Robert Messner ◽  
Christian P. Kubicek
Keyword(s):  
Cell Wall ◽  
Trichoderma Reesei ◽  
Cell Walls ◽  
Cell Wall Protein ◽  
Protoplast Regeneration ◽  
Secretory Proteins ◽  
Soluble Cell ◽  
Cell Wall Biogenesis ◽  
The Individual ◽  
Specific Labelling

The synthesis of constituent polymers of the cell wall by either growing mycelia or regenerating protoplasts of Trichoderma reesei QM 9414 was investigated by following the incorporation of radioactive precursors of the individual polymers (i.e., N[14C] acetyl-glucosamine, [3H] and [14C]glucose, [14C]mannose, and [35S]methionine) into individual fractions. N-Acetyl-glucosamine and glucose were found to become specifically incorporated into cell wall chitin and glucan by both mycelia and regenerating protoplasts, indicating the activity of chitin and glucan synthases in both systems. Cell wall glucan from regenerating protoplasts, however, consisted only of α-glucan and specifically lacked β-glucan which is found in mycelial cell walls. Mannose became metabolized to glucose before its label appeared in the cell wall, and was thus unsuitable for specific labelling. [35S]Methionine was found in a small (< 21 kDa) polypeptide from the first alkali-soluble cell wall fraction, but also in cell wall bound secretory proteins, i. e., β-glucosidase. These results indicate that cell wall biogenesis in T. reesei, in contrast to a report by other authors, is similar to that of other filamentous fungi. Key words: Trichoderma reesei, protoplast regeneration, cell wall polymer, β-glucan, cell wall protein.

A SEROLOGICAL STUDY OF CELL WALLS OF CERTAIN LACTIC ACID BACTERIA

1962 ◽  
Vol 8 (5) ◽  
pp. 629-637
Author(s):  
K. L. Chung ◽  
Roma Z. Hawirko
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Intact Cell ◽  
Synthetic Medium ◽  
Specific Cell ◽  
Intact Cells ◽  
Common Antigens ◽  
Species Specific ◽  
Relationship Of ◽  
The Relationship

From three species of Lactobacillus and three species of Streptococcus, cultured in a synthetic medium, cell walls were isolated following sonic disintegration and purified by washing. Sera against each species were prepared by injecting three rabbits with cell walls, and three with intact cells. Reciprocal agglutination tests were carried out with unabsorbed and absorbed antisera. More kinds of antibodies were detected with cell-wall antisera than with intact-cell antisera. Many species in the two genera shared common antigens. S. faecalis was the exception. Certain antigens believed to be complex haptens in nature reacted with heterologous antisera. Haemagglutination of tanned erythrocytes sensitized with a particulate cell-wall suspension showed fewer cross reactions than agglutination of intact-cell suspensions.The evidence presented shows the possibility of using antisera against species-specific cell-wall antigens for the identification of these species. The relationship of these species is discussed.

scholarly journals The role of plant cell wall encapsulation and porosity in regulating lipolysis during the digestion of almond seeds

2016 ◽  
Vol 7 (1) ◽  
pp. 69-78 ◽  
Author(s):  
Myriam M. L. Grundy ◽  
Frédéric Carrière ◽  
Alan R. Mackie ◽  
David A. Gray ◽  
Peter J. Butterworth ◽  
...  
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Intact Cell ◽  
Lipid Digestion

Intact cell walls of almond prevent lipase penetration thus hindering lipid digestion.

The effect of growth environment on the chloroform–methanol and alkali-extractable cell wall and cytoplasm lipid levels of Mucor rouxii

1975 ◽  
Vol 21 (1) ◽  
pp. 79-84 ◽  
Author(s):  
S. Safe ◽  
J. Caldwell
Keyword(s):  
Fatty Acids ◽  
Cell Wall ◽  
Cell Walls ◽  
Lipid Levels ◽  
Mucor Rouxii ◽  
Growth Environment ◽  
Lipid Fractions ◽  
Filamentous Cell ◽  
The Individual ◽  
Chloroform Methanol

The distribution of chloroform–methanol and alkali-extractable lipids in the cell walls of aerobically grown filamentous cells from Mucor rouxii has been determined. The results have been compared with the corresponding lipid composition of yeast-like cells from M. rouxii, which can be produced in two ways: by growth under anaerobic conditions and by aerobic growth in the presence of 0.22% phenethyl alcohol (PEA). It was observed that in most cases the crude cytoplasmic fraction contained higher levels of several lipids (i.e., squalene, sterols, triterpenes, and fatty acids) than did the corresponding cell walls. The cell walls did, however, contain both "free" (chloroform–methanol extractable) and "bound" (alkali extractable) lipids although the relative amounts were markedly dependent on the cell growth environment. The aerobically grown filamentous cell walls contained higher levels of squalene, sterols, triterpenes, and fatty acids than did aerobically grown yeast-like PEA-induced cell walls and there was also considerable variation in the "free''/"bound" ratios of the various lipid components. The lipid levels in both the cell walls and cytoplasm of the anaerobically grown cells were considerably lower than those of the cells grown under aerobic conditions. In addition, the differences in the growth environment were also reflected in the compositions of the individual lipid fractions from both the cell wall and the cytoplasm fraction.

Application of Scanning UV Microspectrophotometry to Localise Lignins and Phenolic Extractives in Plant Cell Walls

Holzforschung ◽  
2001 ◽  
Vol 55 (6) ◽  
pp. 563-567 ◽  
Author(s):  
G. Koch ◽  
G. Kleist
Keyword(s):  
Cell Wall ◽  
Plant Cell ◽  
Cell Walls ◽  
Individual Cell ◽  
Prunus Serotina ◽  
Plant Cell Walls ◽  
Phyllostachys Edulis ◽  
Woody Tissue ◽  
Analytical Technique ◽  
Uv Microspectrophotometry

Summary The localisation of lignin and phenolic extractives in woody tissue was determined using scanning UV microspectrophotometry. This improved cellular analytical technique enabled direct imaging of the topochemical lignin distribution within individual cell wall layers with a resolution of 0.25 μm2. Selected softwood (Picea abies), hardwood (Fagus sylvatica, Entandrophragma cylindricum, Prunus serotina) and monocotyledon (Phyllostachys edulis) sections of 1 μm thickness were scanned at a fixed wavelength and evaluated with the “APAMOS” software. This approach allowed the distribution pattern of lignins and aromatic extractives within the cell wall to be visualised simultaneously. The method was found to be ideally suited to the study of their subcellular distribution in plant cell walls.

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