A study of the composition and structure of the cell-wall mucopeptide of micrococcus lysodeikticus.
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Cell Wall↗
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Emended description of the genus Trichococcus, description of Trichococcus collinsii sp. nov., and reclassification of Lactosphaera pasteurii as Trichococcus pasteurii comb. nov. and of Ruminococcus palustris as Trichococcus palustris comb. nov. in the low-G+C gram-positive bacteria.
Analyses of 165 rRNA gene sequences, restriction endonuclease digestion fingerprints of 16S-23S intergenic regions, DNA base compositions, fatty-acid profiles, cell-wall chemistry, cell physiology and fermentation end-product composition, along with other biochemical and phenotypic properties, supported the view that Trichococcus flocculiformis EchtT (DSM 2094T), Lactosphaera pasteurii KoTa2T (DSM 2381T), Ruminococcus palustris Z-7189T (DSM 9172T) and an isolate named 'Carnococcus allantoicus' NDP were all very similar and should be merged into a single genus. Detailed characterization of strains Ben 77, Ben 200 and Ben 201 described previously as 'Nostocoida limicola' I, a filamentous bacterium which causes bulking in activated sludge systems, revealed that these strains also belonged to the same genus as T. flocculiformis EchtT, L. pasteurii KoTa2T, R. palustris Z-7189T and 'C allantoicus' NDP. In fact, their shared properties suggested that these strains all belonged to a single species. However, DNA-DNA hybridization data indicated that T. flocculiformis EchtT, all of the 'N. limicola' I isolates and 'C allantoicus' NDP belonged to the same species, whereas L. pasteurii KoTa2T, R. palustris Z-7189T and two new isolates, 37AN3*T and 45AN2, represented three distinct species within the same genus. The priority of the genus name Trichococcus is established and since its validation predates the description of the genus Lactosphaera this name should take precedence. Under certain culture conditions, all of the strains mentioned above could produce chains of cocci. Furthermore, the morphology of T. flocculiformis EchtT could change to a non-filamentous form on certain media. This study proposes that the above strains be reclassified as members of the genus Trichococcus as four species, namely Trichococcus flocculiformis emend. (type strain EchtT = DSM 2094T), Trichococcus pasteurii comb. nov. (type strain KoTa2T = DSM 2381T = ATCC 35945T), Trichococcus collinsii sp. nov. (type strain 37AN3*T = DSM 14526T = ATCC BAA-296T, and Trichococcus palustris comb. nov. (type strain Z-7189T = DSM 9172T).
Bacteria↗
Structure and composition of the alkali-insoluble cell wall fraction of Coprinus macrorhizus var. microsporus.
The alkali-insoluble (R-) fraction from the cell walls of Coprinus macrorhizus var. microsporus is a highly branched glucan, containing alpha-(1 leads to 4), beta-(1 leads to 3), and beta-(1 leads to 6) linkages as shown by methylation, partial acid hydrolysis, and enzymic hydrolysis. The alpha-(1 leads to 4)-linked segments are joined by occasional beta-(1 leads to 3) links as suggested by the identification of 2-O-alpha-glucopyranosyl erythritol in the hydrolysate of the reduced, periodate-oxidized glucan. Hydrolysis of the permethylated glucan gave nearly equimolar amounts of 2,4-di- and 2,3-di-O-methyl-D-glucose. Methylation analysis of the residue from enzymic hydrolysis, the "CORE-fraction," indicated the presence of glucose residues in this fraction linked through positions O1, O3, O4, and O6. Hydrolysates of the R-fraction contained mannose, glucosamine, and amino acids in addition to glucose.
Agaricales↗
[Composition and properties of the outer cell wall layer in Bacillus brevis--the producer of gramicidin S].
Two membrane antigens were found by cross immunoelectrophoresis in the cell walls of Bacillus brevis var. G.-B., R form, which started to synthesize gramicidin S (20 mg per 1 ml of cultural broth). The cell wall contained no membrane components in cells at the beginning of the logarithmic growth phase. The protein with a molecular mass of 100 kDa is a component of the cell wall outer layer. The protein is not digested by trypsin or pronase when it comprises the cell walls of cells synthesizing gramicidin S. In the preparation of isolated cell walls, this protein becomes susceptible to the action of the above proteases only when the peptidoglycan layer is broken down by lysozyme. Electron microscopy of cells treated with proteases and shadowed with a metal revealed that many cells lacked the cytoplasm. Therefore, the outer layer of B. brevis R cell wall contains small regions susceptible to the action of protease along with regions composed of the 100 kDa protein and resistant to these enzymes. It is possible that the small regions contain membrane components.
Antigens, Bacterial↗
[Chemical composition of a preparation of the cell walls of Corynebacterium diphtheriae].
The chemical characteristics of 6 batches of the preparation, obtained from the cell wall of C. diphtheriae grown in liquid and solid culture media, with respect to their content of nitrogen, hexoses, pentoses, total amino sugars, lipids and to the possible admixture of nucleic acids are presented. From the results of the chemical analysis of these batches their standardization according to the ratio of total amino sugars and pentoses to total nitrogen in C. diphtheriae cell-wall preparation is proposed.
Cell Wall↗
Homology of aminoacid composition of structural glycoproteins, transplantation antigens, cell wall glycoproteins, and streptococcus A cell membrane.
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Amino Acid Sequence↗
The lipopeptide antimycotic, cilofungin modulates the incorporation of glucan-associated proteins into the cell wall of Candida albicans.
The effect of the beta 1-3 glucan synthase inhibitor, cilofungin, on the incorporation of 35S-methionine-labelled glucan associated proteins (GAP) in the cell wall of Candida albicans was investigated in a susceptible strain C. albicans 3153 and resistant strain C. albicans CA-2. Cilofungin exerted a marked effect on the GAP composition of the cell wall at 0.25 mg/L, a concentration which reduced beta 1-3 glucan synthesis by approximately 50% and also inhibited the growth of the susceptible strain C. albicans 3153. A 46 kDa protein was present in large amounts in C. albicans 3153 but not in strain CA-2. This protein was probably not mannosylated and its incorporation was greatly reduced by cilofungin. In addition, a well defined 34 kDa protein was identified together with a distinct band of high molecular mass polydisperse material of between 65 and 96 kDa and another of > 200 kDa. These proteins were strongly reactive to concanavalin A indicating that they were mannosylated, and treatment with cilofungin caused an increase in their production which was also confirmed by immunoblotting with rabbit anti-Candida serum. In contrast, exposure of the drug-resistant strain CA-2 to cilofungin did not result in changes in the composition of the GAP constituents. Only the mannosylated proteins of 34 kDa and the high molecular mass polydisperse material 65-96 kDa were present in the cell wall. The pulse-chase labelling experiments showed that the 46 kDa protein was the first of the GAPs to be incorporated into the cell wall, and that this was suppressed in the presence of cilofungin whereas there was a concomitant increase in the incorporation of the 34 kDa and the high-molecular weight polydisperse material. Thus, cilofungin causes a profound imbalance in GAP incorporation into the growing cell wall which is possibly related to changes in the amount and type of glucan being synthesized at sub-inhibitory concentrations of the antimycotic.
Antifungal Agents↗
Structure and biosynthesis of fungal cell walls: methodological approaches.
Fungal cell walls possess a characteristic chemical composition differentiating fungal cells from other cell types. For this reason, the mechanisms involved in cell-wall formation represent a potential target for selective antifungal drugs. Understanding the structure and biosynthesis of fungal cell walls opens the ways for design of effective drugs for treating fungal diseases. This article reviews the history methods employed in chemical and structural analysis of fungal cell walls and in studies concerning their formation.
Biochemistry↗
GPI7 affects cell-wall protein anchorage in Saccharomyces cerevisiae and Candida albicans.
Glycosylphosphatidylinositol (GPI)-anchoring represents a mechanism for attaching proteins to the cell surface of all eukaryotic cells. Two localizations of GPI proteins have been observed in the yeasts Saccharomyces cerevisiae and Candida albicans: plasma membrane and cell wall. The signals and the mechanisms involved in this differential targeting are presently not well understood. Here several cell-wall-related phenotypes of a gpi7/las21 deletion are described, where GPI7/LAS21 encodes a GPI-anchor-modifying activity. In both organisms, the structure and composition of the cell wall was modified, with a clear increase in chitin deposition. Cell-wall-targeted proteins accumulated in the growth medium, whereas the protein content of the cell wall decreased significantly, suggesting inefficiency of the covalent linkage. The level of plasma-membrane-targeted GPI proteins was not affected. Sequence analyses revealed that gene families involved in the addition of phosphoethanolamines to the core GPI anchor are highly conserved between eukaryotes, with the exception of the Gpi7 family which seems to be fungus-specific. These data are compatible with the notion that the phosphoethanolamine added by Gpi7 protein to the GPI anchor is a key factor in the covalent linkage of cell-wall proteins to fungal cell-wall components.
Amino Acid Sequence↗
Fractionation of cell-wall preparations from grass leaves by centrifuging in non-aqueous density gradients.
1. Dried preparations of cell walls from perennial-ryegrass (Lolium perenne) and Italian-ryegrass (L. multiflorum) leaves were suspended in mixtures of carbon tetrachloride with light-petroleum (b.p. 45--50 degrees C) or alcohols and layered on density gradients formed from the same solvents. 2. On centrifugation, the cell walls become distributed throughout a suitably chosen gradient. Fractions corresponding to various regions of the gradient were separated, examined under the microscope and analysed. 3. Cell-wall preparations made from leaf material ground in liquid N2, or in a triple roll mill, showed considerable heterogeneity in particle size, and their behaviour in the density gradient was variable, although there was a general indication that walls derived from vascular bundles were less dense than those from sclerenchyma. 4 Treatment in a vibratory ball mill decreased the size of the particles and produced a more uniform material, but made it impossible to distinguish the origins of the particles. This material behaved more reproducibly in the density gradient. 5. Some fractionations were also made by successive centrifugation in media of increasing relative density. 6. Analyses of the fractions obtained by each method indicated that the less dense had a greater proportion of xylose in the polysaccharide components, and higher contents of acetyl groups and lignin, confirming the close relationship between these components in plant cell walls. 7. The results show that there are differences in polysaccharide composition between the cell-wall types in the grass leaf, the vascular tissue being richer in hemicellulose relative to cellulose than the sclerenchyma.
Butanols↗
Cell wall alterations associated with the hyperproduction of extracellular enzymes in Neurospora crassa.
A pleiotropic mutation in Neurospora (exo-1), which confers derepression of alpha-amylase, glucoamylase, beta-fructofuranosidase, and trehalase, appears to also affect the composition of the cell wall. Segregants resulting from the backcross of exo-1 to the wild-type strain from which it derived are altered in the ratio of galactosamine to glucosamine in hydrolysates of isolated cell walls. Conidial cell walls exhibit a marked decrease in the amount of galactosamine in both exo-1 and exo-1(+) strains. Increased levels (approximately sevenfold) of amylase are found in conidia of exo-1, as compared with those of exo-1(+).
Amylases↗
Effects of microgravity on the structure and function of plant cell walls.
The structural, biochemical, cytofluorimetric and electron cytochemical features of the cell walls of higher plants grown under weightlessness and simulated microgravity are described. Space flight and laboratory clinostatic experiments with plants show that the ultrastructure of the cell wall, its polysaccharide composition, and metabolic organization depend on the type of tissue and the duration of weightlessness. Horizontal clinostating that reproduced the biological effects of microgravity on cell walls showed that the structure of the external walls of the epidermis of aboveground organs is very sensitive to microgravity. Various responses occur in the primary and secondary walls under weightlessness and clinorotation: rearrangement of cell walls and organelles and changes in the content of cellulose, lignin, callose, and hemicelluloses. It is shown that plant cell wall changes under microgravity are connected with changes in cellulase, pectinase, and peroxidase activity and a change in the calcium balance in the cytoplasm and apoplast.
Calcium↗
Membrane alteration and the formation of metachromatic granules in Escherichia coli treated with p-fluorophenylalanine.
The effect of p-fluorophenylalanine (FPA) on growing cultures of Escherichia coli was studied with regard to the composition and morphology of the cell envelope. A cell wall fraction was prepared by autolysis in hypertonic medium, and the resulting spheroplasts were osmotically lysed to obtain a cytoplasmic membrane fraction. Incorporation of labeled phenylalanine, FPA, and N-acetylglucosamine into both fractions of FPA-inhibited cells suggested that the composition of the membrane changed with time, whereas that of the cell wall remained relatively constant. Amino acid analysis revealed changes in the composition of the membrane fraction after FPA inhibition. Electron micrographs of shadowed cells and membranes revealed the presence of electron-dense metachromatic granules during the early stages of FPA inhibition.
Amino Acids↗
Analysis of the cell wall of five strains of Myocbacterium tuberculosis BCG and of an attenuated human strain, W 115.
The chemical composition of the deproteinized, delipidated cell walls of five strains of BCG and of an attenuated human strain of Mycobacterium tuberculosis has been established, with special focus on their poly-L-glutamic acid content. All the cell walls have the same overall composition. Their poly-L-glutamic acid content varies from 0 to 4.6%. A correlation between the poly-L-glutamic acid content and the biological properties of the BCG strains reported in the literature could not be established.
Amino Acids↗
Kinetics of cell-wall digestion of orchardgrass and alfalfa silages treated with cellulase and formic acid.
The objectives of this study were to determine the effects of cellulase (from Trichoderma longibrachiatum) combined with formic acid, applied before ensiling, on the subsequent concentration and composition of the cell wall and on the extent and rate of in situ cell-wall digestion of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.). Treated and control forages of both plant species were ensiled for at least 60 d before being ruminally digested by two fistulated cows. Analyses of NDF, ADF, and acid detergent lignin were conducted sequentially on original and digested samples. Data were fitted with a first-order, nonlinear model to estimate extents and rates of digestion of potentially digestible NDF, cellulose, and hemicellulose. The concentration of indigestible residue and the discrete lag time before digestion were also determined for the cell-wall components. After ensiling, the mean NDF concentration of treated silages was 19% lower than that of control silages; the effect was greater for orchardgrass than for alfalfa. The extent of digestion of NDF, cellulose, and hemicellulose, respectively, was 33, 37, and 27% lower for treated silages than for control silages. Treatment effects on the extent of digestion varied between plant species. Cellulose from treated orchardgrass was digested 19% more slowly than cellulose from the control silage. Indigestible residue concentrations of NDF, cellulose, and hemicellulose, respectively, were 7, 8, and 7% lower in treated silages than in control silages. Thus, extensive cell-wall degradation by cellulase during ensiling resulted in less digestible cell-wall material for ruminal digestion but greater total cell-wall degradation, including that during ensiling and ruminal incubation, especially during early digestion in the rumen.
Cell Wall↗
[Study on the application of quantitative analysis of cell-wall amino acids in actinomycetes classification].
This paper has collected 70 recognized type strains of actinomycete from home and abroad Microbiological culture collection centers. Adopting thin-layer chromatography and thin-layer chromatogram scanner methods to quantitatively analyze the composition of cell-wall amino acids in actinomycete. It has made comparison between quantitative results and qualitative ones. After a thorough discussion, a revision is made on the standard of dividing cell-wall types in actinomycete chemical classification, which has provided some original suggestion in hope of improving the research of actinomycete chemical classification.
Actinobacteria↗
Five novel species of the genus Nocardiopsis isolated from hypersaline soils and emended description of Nocardiopsis salina Li et al. 2004.
Five novel Nocardiopsis strains isolated from hypersaline soils in China were subjected to a polyphasic analysis to determine their taxonomic position. All of the novel isolates could grow on agar plates at NaCl concentrations of up to 18 % (w/v), with optimum growth at 5-8 %. The DNA G+C contents of the novel strains ranged from 67.9 to 73.2 mol%. The morphological and chemotaxonomic characteristics of the isolates matched those described for members of the genus Nocardiopsis. Based on their 16S rRNA gene sequence analysis, DNA-DNA hybridization values and phenotypic characteristics, including the composition of cell-wall amino acids and sugars, menaquinones, polar lipids and cellular fatty acids, the isolates are proposed as representing five novel species of the genus Nocardiopsis. The novel species are proposed as Nocardiopsis gilva sp. nov. [type strain YIM 90087T (=KCTC 19006T=CCTCC AA 2040012T=DSM 44841T)], Nocardiopsis rosea sp. nov. [type strain YIM 90094T (=KCTC 19007T=CCTCC AA 2040013T=DSM 44842T), Nocardiopsis rhodophaea sp. nov. [type strain YIM 90096T (=KCTC 19049T=CCTCC AA 2040014T=DSM 44843T), Nocardiopsis chromatogenes sp. nov. [type strain YIM 90109T (=KCTC 19008T=CCTCC AA 2040015T=DSM 44844T) and Nocardiopsis baichengensis sp. nov. [type strain YIM 90130T (=KCTC 19009T=CCTCC AA 2040016T=DSM 44845T). On the basis of the chemotaxonomic data, the description of the recently described species Nocardiopsis salina Li et al. 2004 is emended.
Actinomycetales↗
[Immunological study of lysates from the cell wall of group A Streptococcus].
The chemical composition and presence of immunogenic components in the lysates of the cell walls of group A Streptococcus, type M29, were studied. The lysates were prepared with the use of muramidase. Fc-Receptors were detected in the lysates. Within the first 30 minutes of cell wall lysis by muramidase, 4 times higher amounts of the protein reacting with fibrinogen excreted than in the subsequent 4 hours. The lysates contained immunogenic proteins. Fraction III isolated by chromatography of the 30-minute lysate on DEAE-trisacryl formed a single precipitation band with lysate antiserum. The lysate Fraction IV forming three precipitation bands contained a protein not specific of the type. The protein was identical to the protein antigen from Triton X-100 extracts of group A Streptococcus, types M1, M12 and M29. The group-specific polysaccharide was detected in the lysate Fraction I and Fraction II of the 4-hour lysate.
Bacterial Proteins↗