Rapid isolation method for lipopolysaccharide and lipid A from gram-negative bacteria.

@article{Yi2000RapidIM,
  title={Rapid isolation method for lipopolysaccharide and lipid A from gram-negative bacteria.},
  author={E C Yi and Murray Hackett},
  journal={The Analyst},
  year={2000},
  volume={125 4},
  pages={
          651-6
        }
}
A fast, convenient extraction method for lipopolysaccharide (LPS), using a commercial RNA isolating reagent, allows the isolation of LPS or lipid A from low milligram (dry weight) quantities of bacterial cells. The method avoids the use of specialized equipment and has been used for processing relatively large numbers of samples. The major components of the commercial RNA isolating reagent, Tri-Reagent, are phenol and guanidinium thiocyanate in aqueous solution. The bacterial cell membranes are… 
View on PubMed
lib3.dss.go.th
197 Citations
Highly Influential Citations
10
Background Citations
9
Methods Citations
71

Figures and Tables from this paper

197 Citations

Citation Type

Citation Type

Isolation and chemical characterization of lipid A from gram-negative bacteria.
TLDR
T tandem MS protocols for analyzing lipid A molecular species using electrospray ionization coupled to collision induced dissociation (CID) and newly employed ultraviolet photodissociation (UVPD) methods are described, allowing for unequivocal determination of chemical structure, paramount to characterization of lipid A molecules that contain unique or novel chemical modifications.
Microextraction of bacterial lipid A: easy and rapid method for mass spectrometric characterization Published, JLR Papers in Press, June 1, 2005. DOI 10.1194/jlr.D500014-JLR200
TLDR
This method using hot ammonium-isobutyrate solvent was applied to small quantities of cells and proved to be indispensable when a rapid characterization of lipid A structure by mass spectrometry was required.
A rapid one-step method for the characterization of membrane lipid remodeling in Francisella using matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry.
TLDR
This analysis showed that the bacterial membranes remodeled rapidly to adapt to changes in environmental growth conditions and may be important for Francisella pathogenesis.
A two-step method for extraction of lipopolysaccharide from Shigella dysenteriae serotype 1 and Salmonella typhimurium: An improved method for enhanced yield and purity.
Structural characterization of bacterial lipopolysaccharides with mass spectrometry and on- and off-line separation techniques.
TLDR
This review examines the tandem mass spectrometric methods and protocols for the analyses of lipid A, the endotoxic principle of LPS, and compares and evaluates the different ionization techniques in view of their use in intact R- and S-type LPS and lipid A studies.
Purification of Porphyromonas gingivalis polysaccharide free lipopolysaccharide using Sephacryl S-200 high resolution chromatography
TLDR
The purification protocol allowed us to obtain a highly purified P. gingivalis LPS which could be used in future tests to evaluate its behavior in vitro and in vivo and elucidate its function, as well as to obtain LPS from other períodontopatic bacteria to address the association of períODontal disease with cardiovascular diseases.
Mass Spectrometry for Profiling LOS and Lipid A Structures from Whole-Cell Lysates: Directly from a Few Bacterial Colonies or from Liquid Broth Cultures.
TLDR
A fast and easy micromethod suitable for extracting small quantities of LOS and a slightly modified micrometshod for the detection of the lipid A constituents of the LPSs from bacteria grown in different culture media are described and the structures with mass spectrometry are evaluated.
The outer membrane of Gram-negative bacteria: lipid A isolation and characterization.
The isolation and characterization of the lipid A domain of lipopolysaccharide (LPS) are important methodologies utilized to gain understanding of the Gram-negative cell envelope. Here, we describe…
A combined fermentative-chemical approach for the scalable production of pure E. coli monophosphoryl lipid A
TLDR
The reported method for the large-scale obtainment of monophoshoryl lipid A from the fed-batch fermentation broth of a recombinant strain of E. coli may permit the access to novel semisynthetic lipid A immunoadjuvant candidates.
Analysis of Lipid a Isolated from Brucella ovis R-Lipopolysaccharide
TLDR
The fatty acid profile found in lipid A is very rich in palmitic acid and is able to identify a location of the cyclopropane carboxylic acids described for this bacteria.
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 17 REFERENCES
Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains
TLDR
A LPS isolation procedure which is effective in extracting both smooth and rough LPS in high yields and with a high degree of purity is developed and demonstrated by comparing LPS preparations obtained from wild-type and mutant strains of P. aeruginosa.
Improved techniques for the preparation of bacterial lipopolysaccharides.
TLDR
Pretreatment of cells with lysozyme in the presence of ethylenediaminetetraacetic acid was the most efficient method in terms of lipopolysaccharide yield and ease of preparation.
A colorimetric estimation of lipopolysaccharides
Degradative Effect of Phenol on Endotoxin and Lipopolysaccharide Preparations from Serratia marcescens
It has been established that the well-known deproteinizing action of hot 45% aqueous phenol on whole cells or isolated and purified endotoxin of Serratia marcescens 08 is caused by the cleavage of a…
A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels.
Lipid A-like molecules that antagonize the effects of endotoxins on human monocytes.
Detergent-accelerated hydrolysis of bacterial endotoxins and determination of the anomeric configuration of the glycosyl phosphate present in the "isolated lipid A" fragment of the Bordetella pertussis endotoxin.
Mechanism of assembly of the outer membrane of Salmonella typhimurium. Site of synthesis of lipopolysaccharide.
A novel Escherichia coli lipid A mutant that produces an antiinflammatory lipopolysaccharide.
TLDR
A unique screen was used to identify mutations in Escherichia coli lipid A biosynthesis that result in a decreased ability to stimulate E-selectin expression by human endothelial cells, and the cloned MSbB gene was able to functionally complement the msbB mutant, restoring both the LPS to its native composition and the ability of the strain to stimulate immune cells.
IL-1 induction-capacity of defined lipopolysaccharide partial structures.
TLDR
It is concluded that lipid A is the main portion of LPS responsible for induction of IL-1, and that specific activation- and-or binding-mechanisms are involved in stimulation of cells with LPS and/or lipid A.
...
1
2
...