Chemical analysis of isolated cell walls of Gram-positive bacteria and determination of the cell wall to cell mass ratio

1997 ◽  
Vol 28 (2) ◽  
pp. 147-157 ◽  
Author(s):  
Albert van der Wal ◽  
Willem Norde ◽  
Bernd Bendinger ◽  
Alexander J.B Zehnder ◽  
Johannes Lyklema
1989 ◽  
Vol 35 (8) ◽  
pp. 749-753 ◽  
Author(s):  
Denis Leclerc ◽  
Alain Asselin

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.


1999 ◽  
Vol 43 (9) ◽  
pp. 2314-2316 ◽  
Author(s):  
Arnold H. Horwitz ◽  
Robert E. Williams ◽  
Pei-Syan Liu ◽  
Rossana Nadell

ABSTRACT Bactericidal/permeability-increasing protein (BPI) inhibited growth of cell wall-deficient Acholeplasma laidlawii and L forms of certain strains of Staphylococcus aureus andStreptococcus pyogenes. However, the same strains ofS. aureus and S. pyogenes with intact cell walls were not susceptible to the growth-inhibitory effects of BPI.


1997 ◽  
Vol 9 (1-2) ◽  
pp. 81-100 ◽  
Author(s):  
Albert van der Wal ◽  
Willem Norde ◽  
Alexander J.B. Zehnder ◽  
Johannes Lyklema

1998 ◽  
Vol 64 (8) ◽  
pp. 3059-3062 ◽  
Author(s):  
Elena Bidnenko ◽  
Carine Mercier ◽  
Josselyne Tremblay ◽  
Patrick Tailliez ◽  
Saulius Kulakauskas

ABSTRACT Fluorescent in situ hybridization (FISH) is now a widely used method for identification of bacteria at the single-cell level. With gram-positive bacteria, the thick peptidoglycan layer of a cell wall presents a barrier for entry of horseradish peroxidase (HRP)-labeled probes. Therefore, such probes do not give any signal in FISH unless cells are first treated with enzymes which hydrolyze the peptidoglycan. We explored this feature of FISH to detect cells which have undergone permeabilization due to expression of autolytic enzymes. Our results indicate that FISH performed with HRP-labeled probes provides a sensitive method to estimate the states of cell walls of individual gram-positive bacteria.


Biochemistry ◽  
1965 ◽  
Vol 4 (10) ◽  
pp. 2237-2244 ◽  
Author(s):  
J. M. Ghuysen ◽  
L. Dierickx ◽  
M. Leyh-Bouille ◽  
J. L. Strominger ◽  
E. Bricas ◽  
...  

1971 ◽  
Vol 17 (3) ◽  
pp. 421-424 ◽  
Author(s):  
H. E. Gilleland Jr. ◽  
I. L. Roth ◽  
R. G. Eagon

An ultrastructural study of a Gram-variable coccus was carried out. The cell wall of this microorganism was composed of an inner peptidoglycan layer, a middle electron-transparent compartment, and an undulating trilayered outer membrane. This microorganism also possessed numerous mesosomes which were simple bulb-like invaginations of the cytoplasmic membrane. The mechanism of cellular division involved the formation of a septum by the cytoplasmic membrane and the inner layer of the cell wall. Membranous structures were associated with the developing septum throughout the process. The outer membrane of the cell wall did not invaginate with the inner layer but reformed as the completed septum began to split. In isolated cell wall preparations, no 2-keto-3-deoxyoctonate or heptose could be detected. It is suggested that the Gram-variable cocci previously classified as micrococci may represent a group that is intermediate between true Gram-negative and Gram-positive bacteria.


1997 ◽  
Vol 161 ◽  
pp. 491-504 ◽  
Author(s):  
Frances Westall

AbstractThe oldest cell-like structures on Earth are preserved in silicified lagoonal, shallow sea or hydrothermal sediments, such as some Archean formations in Western Australia and South Africa. Previous studies concentrated on the search for organic fossils in Archean rocks. Observations of silicified bacteria (as silica minerals) are scarce for both the Precambrian and the Phanerozoic, but reports of mineral bacteria finds, in general, are increasing. The problems associated with the identification of authentic fossil bacteria and, if possible, closer identification of bacteria type can, in part, be overcome by experimental fossilisation studies. These have shown that not all bacteria fossilise in the same way and, indeed, some seem to be very resistent to fossilisation. This paper deals with a transmission electron microscope investigation of the silicification of four species of bacteria commonly found in the environment. The Gram positiveBacillus laterosporusand its spore produced a robust, durable crust upon silicification, whereas the Gram negativePseudomonas fluorescens, Ps. vesicularis, andPs. acidovoranspresented delicately preserved walls. The greater amount of peptidoglycan, containing abundant metal cation binding sites, in the cell wall of the Gram positive bacterium, probably accounts for the difference in the mode of fossilisation. The Gram positive bacteria are, therefore, probably most likely to be preserved in the terrestrial and extraterrestrial rock record.


Author(s):  
B.K. Ghosh

Periplasm of bacteria is the space outside the permeability barrier of plasma membrane but enclosed by the cell wall. The contents of this special milieu exterior could be regulated by the plasma membrane from the internal, and by the cell wall from the external environment of the cell. Unlike the gram-negative organism, the presence of this space in gram-positive bacteria is still controversial because it cannot be clearly demonstrated. We have shown the importance of some periplasmic bodies in the secretion of penicillinase from Bacillus licheniformis.In negatively stained specimens prepared by a modified technique (Figs. 1 and 2), periplasmic space (PS) contained two kinds of structures: (i) fibrils (F, 100 Å) running perpendicular to the cell wall from the protoplast and (ii) an array of vesicles of various sizes (V), which seem to have evaginated from the protoplast.


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