scholarly journals The action of dilute aqueous NN-dimethylhydrazine on bacterial cell walls

1969 ◽  
Vol 113 (1) ◽  
pp. 183-189 ◽  
Author(s):  
J. C. Anderson ◽  
A. R. Archibald ◽  
J Baddiley ◽  
M. J. Curtis ◽  
N. Barbara Davey
Keyword(s):  
Molecular Weight ◽  
Free Radicals ◽  
High Molecular Weight ◽  
Bacterial Cell ◽  
Cell Walls ◽  
Cross Linking ◽  
Gram Positive ◽  
Teichoic Acids ◽  
Gram Positive Bacteria ◽  
Almost All

1. Walls of certain Gram-positive bacteria dissolved on incubation with dilute aqueous NN-dimethylhydrazine in the presence of air, by a reaction that probably involves free radicals. 2. Under the conditions described, the soluble products from the peptidoglycan were almost all non-diffusible. After brief incubation of walls of some organisms with reagent, part of the peptidoglycan component was obtained as a high-molecular-weight gel, the viscosity of which was rapidly decreased by incubation with lysozyme. 3. The extent to which peptidoglycan dissolved varied with different organisms, depending possibly on the extent of cross-linking, but the nature of the bonds that were destroyed has not been established. 4. Teichoic acids and polysaccharides were solubilized by this treatment and could be isolated in high overall yield. 5. The procedure is valuable in the examination of the distribution of heteropolymers in walls, and has been used to show that the polysaccharide present in walls of Lactobacillus arabinosus 17–5 is phosphorylated and may account for 20% of the total phosphate of the wall.

scholarly journals Sir James Baddiley. 15 May 1918 — 19 November 2008

2010 ◽  
Vol 56 ◽  
pp. 3-23
Author(s):  
J. Grant Buchanan
Keyword(s):  
Organic Chemistry ◽  
Adenosine Triphosphate ◽  
Bacterial Cell ◽  
Cell Walls ◽  
Structure And Function ◽  
Cambridge University ◽  
Teichoic Acids ◽  
Gram Positive Bacteria ◽  
Organic Chemist ◽  
And Function

James Baddiley was a biochemist who used the methods and insight of the organic chemist to answer important questions in biology, notably coenzyme structure and the structure and function of bacterial cell walls. A graduate of Manchester University, he moved to Cambridge in 1944 with A. R. Todd, where he synthesized adenosine triphosphate, the nucleotide concerned with essential energy transformations in all forms of life. As an independent researcher at the Lister Institute in London he elucidated the structure of coenzyme A and other coenzymes. He was appointed Professor of Organic Chemistry in Newcastle, where the exploration of the structures of two cytidine nucleotides led to the discovery of the teichoic acids, major components of the cell walls and membranes of Gram-positive bacteria. These discoveries were extended to cover the structures, biosynthesis, function and immunology of the teichoic acids. Baddiley became Professor of Chemical Microbiology in 1977. Moving to Cambridge after his retirement, he was able to continue his researches in the Department of Biochemistry. He was elected a Fellow of Pembroke College and as an elder statesman undertook extensive committee work, often as chairman, both in Cambridge University and nationally. He was knighted in 1977.

scholarly journals Selective extraction of polymers from cell walls of Gram-positive bacteria (Short Communication)

1973 ◽  
Vol 131 (3) ◽  
pp. 619-621 ◽  
Author(s):  
Jiri G. Pavlik ◽  
Howard J. Rogers
Keyword(s):  
Bacillus Licheniformis ◽  
Cell Walls ◽  
Short Communication ◽  
Selective Extraction ◽  
Gram Positive ◽  
Teichoic Acids ◽  
Gram Positive Bacteria ◽  
Teichuronic Acid

Brief heating of Bacillus Licheniformis cell walls at 100°C in aqueous buffers of pH3.0–4.0 removes some polymers but not others from the mucopeptides. For example, relatively undegraded teichuronic acid can be extracted at 100°C in 20min at pH3.0 whereas the teichoic acids are not removed. Similar specificity can be shown with walls from three other species of micro-organism.

Detection of bacterial cell wall hydrolases after denaturing polyacrylamide gel electrophoresis

1989 ◽  
Vol 35 (8) ◽  
pp. 749-753 ◽  
Author(s):  
Denis Leclerc ◽  
Alain Asselin
Keyword(s):  
Cell Wall ◽  
Bacterial Cell ◽  
Gel Electrophoresis ◽  
Cell Walls ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Cell Wall Hydrolases

Cell walls from various Gram-positive bacteria were incorporated at a concentration of 0.2% (w/v) into polyacrylamide gels as a substrate for detection of cell wall hydrolases. Bacterial extracts from crude cell wall preparations were denatured with sodium dodecyl sulfate and 2-mercaptoethanol and subjected to denaturing polyacrylamide gel electrophoresis in gels containing bacterial cell walls. After renaturation in the presence of purified and buffered 1% (v/v) Triton X-100, cell wall hydrolases were visualized as clear lytic zones against the opaque cell wall background. One to fifteen bands with lytic activity could be detected, depending on bacterial extracts and on the nature of the cell walls incorporated into gels. Crude cell wall extracts were the best source of cell wall hydrolases from various Gram-positive bacteria such as Clostridium perfringens (15 bands), Micrococcus luteus (1 band), Bacillus megaterium (4 bands), Bacillus sp. (6 bands), B. cereus (3 bands), B. subtilis (7 bands), Staphylococcus aureus (13 bands), Streptococcus faecalis (3 bands), and Strep. pyogenes (5 bands). Molecular masses of cell wall hydrolases ranged from 17 to 114.6 kDa. Lytic activities against cell walls of Corynebacterium sepedonicum (Clavibacter michiganense pv. sepedonicum) could be shown with the cell wall extracts of Strep. pyogenes (45.7 kDa), Strep. faecalis (67 kDa), B. megaterium (67 kDa), and Staph. aureus (67 kDa).Key words: autolysins, electrophoresis, hydrolases, muramidases, peptidoglycan.

scholarly journals Closing Clostridium botulinum Group III Genomes Using Long-Read Sequencing

2021 ◽  
Vol 10 (22) ◽  
Author(s):  
Cedric Woudstra ◽  
Tommi Mäklin ◽  
Yagmur Derman ◽  
Luca Bano ◽  
Hanna Skarin ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Clostridium Botulinum ◽  
Gram Positive ◽  
High Molecular Weight Dna ◽  
Content Type ◽  
Group Iii ◽  
Gram Positive Bacteria ◽  
Complete Genomes ◽  
Long Read

Clostridium botulinum group III is the anaerobic Gram-positive bacteria producing the deadly neurotoxin responsible for animal botulism. Here, we used long-read sequencing to produce four complete genomes from Clostridium botulinum group III neurotoxin types C, D, C/D, and D/C. The protocol to obtain high-molecular-weight DNA from C. botulinum group III is described.

scholarly journals Packed Like Sardines – How Surface Crowdedness Impacts Accessibility to Peptidoglycan of Staphylococcus aureus

2020 ◽  
Author(s):  
Noel J. Ferraro ◽  
Marcos M. Pires
Keyword(s):  
Staphylococcus Aureus ◽  
Bacterial Cell ◽  
Dense Layer ◽  
Bacterial Cells ◽  
Gram Positive ◽  
Teichoic Acids ◽  
Gram Positive Bacteria ◽  
Wide Range ◽  
Small Molecule Therapeutics ◽  
Live Bacterial Cells

AbstractBacterial cell walls are essential barriers that protect bacteria against the onslaught of potentially lethal molecules from the outside. Small molecule therapeutics, proteins from bacterial foes, and host immune proteins must navigate past a dense layer of bacterial biomacromolecules (e.g., capsular proteins, teichoic acids, and anchored proteins) to reach the peptidoglycan (PG) layer of Gram-positive bacteria. A subclass of molecules (e.g., antibiotics with intracellular targets) must also permeate through the PG (in a molecular sieving manner) to reach the cytoplasmic membrane. In the case of Staphylococcus aureus (S. aureus), teichoic acids are the major biopolymers that decorate bacterial cell surfaces. Despite the biological and therapeutic importance of surface accessibility, systematic analyses in live bacterial cells have been lacking. We describe a novel live cell fluorescence assay that reports on the permeability of molecules to and within the PG scaffold. The assay has robust reproducibility, is readily adoptable to any Gram-positive organism, and is compatible with high-throughput sample processing. Analysis of the factors controlling permeability to S. aureus and the methicillin resistant MRSA revealed that molecular flexibility plays a central role in molecular permeability. Moreover, teichoic acids impeded permeability of molecules of a wide range of sizes and chemical composition.

Interaction of Streptococcus mutans Glucosyltransferases with Teichoic Acids

1980 ◽  
Vol 29 (2) ◽  
pp. 376-382
Author(s):  
H. K. Kuramitsu ◽  
L. Wondrack ◽  
M. McGuinness
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Streptococcus Mutans ◽  
Teichoic Acid ◽  
Lipoteichoic Acid ◽  
Water Soluble ◽  
Heat Inactivation ◽  
Teichoic Acids ◽  
Insoluble Glucan

The Streptococcus mutans GS5 glucosyltransferase activities (both water-soluble and -insoluble glucan-synthesizing fractions) were inhibited by purified lipoteichoic acid. In vitro sucrose-dependent colonization of smooth surfaces by strain GS5 was also markedly reduced in the presence of the amphipathic molecules. The inhibition of soluble glucan synthesis by lipoteichoic acid appeared to be competitive with respect to both sucrose and primer dextran T10. These inhibitory effects were dependent on the presence of the fatty acid components of lipoteichoic acid since deacylated lipoteichoic acids did not inhibit glucosyltransferase activity. However, the deacylated molecules did interact with the enzymes since deacylated lipoteichoic acid partially protected the enzyme activity against heat inactivation and also induced the formation of high-molecular-weight enzyme complexes from the soluble glucan-synthesizing fraction. The presence of teichoic acid in high-molecular-weight aggregates of glucosyltransferase isolated from the culture fluids of strain GS5 was suggested by the detection of polyglycerophosphate in these fractions. In addition to strain GS5, two other organisms containing polyglycerophosphate teichoic acids, Lactobacillus casei and Lactobacillus fermentum , were demonstrated to bind glucosyltransferase activity. These results are discussed relative to the potential role of teichoic acid-glucosyltransferase interactions in enzyme binding to the cell surface of S. mutans and the formation of high-molecular-weight enzyme aggregates in the culture fluids of the organism.

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