Abstract
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 phenol-sensitive linkage within the lipid moiety. As a result of this degradation, both the lipopolysaccharide and simple protein fragments retained a part of the lipid moiety. Although not proceeding at the same fast rate as the cleavage of the lipid moiety, such phenol treatment also caused a partial hydrolysis of the O-specific side chain and ester-bound fatty acids. Hydrolysis of the O-specific side chain accounted for 5% of the lipopolysaccharide and that of ester-bound fatty acids accounted for 11% of the total fatty acid content after 60 min of treatment. It is suggested that the presence of these degradation products is one of the main causes of the heterogeneity of endotoxin and lipopolysaccharide preparations.
MeSH Terms
Animals
Chromatography, Paper
Electrophoresis, Paper
Endotoxins/analysis,isolation & purification,toxicity
Hydrolysis
Immunodiffusion
Immunoelectrophoresis
Lipopolysaccharides/analysis,isolation & purification,toxicity
Mice
Mice, Inbred BALB C
Phenols/pharmacology
Polysaccharides, Bacterial/analysis,isolation & purification,toxicity
Serratia marcescens
Solvents
Spectrophotometry, Infrared
Ultracentrifugation
Chemicals
Endotoxins
Lipopolysaccharides
Phenols
Polysaccharides, Bacterial
Solvents
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tsang J C
Wang C S
Alaupovic P
References (27)
27 references, click to expand
-
Biochemical studies on lipopolysaccharides of Salmonella R mutants. 6. Investigations on the structure of the lipid A component.
Eur J Biochem. 1969 Jan;7(3):370-9
PMID: 4307215
-
Relation of structure to function in bacterial O-antigens. IV. Fractionation of the components.
Ann N Y Acad Sci. 1966 Jun 30;133(2):586-603
PMID: 4960347
-
Recent results on the biochemistry of the cell wall lipopolysaccharides of Salmonella bacteria.
Angew Chem Int Ed Engl. 1970 Sep;9(9):649-63
PMID: 4990677
-
A critical examination of the Nelson-Somogyi method for the determination of reducing sugars.
Anal Biochem. 1966 Jun;15(3):373-81
PMID: 5956114
-
A SIMPLE METHOD FOR PREPARING ANTIGENIC SUBSTANCES FROM THE TYPHOID BACILLUS.
Science. 1940 Aug 16;92(2381):155-6
PMID: 17815395
-
Glucosamine values of muramic acid and other amino-sugars by the Elson and Morgan method.
Nature. 1960 Jul 2;187:38-40
PMID: 13835093
-
Qualitative and quantitative colorimetric determination of heptoses.
J Biol Chem. 1953 Oct;204(2):983-97
PMID: 13117875
-
Structural heterogeneity in the lipopolysaccharide of Aerobacter aerogenes NCTC 243.
Biochemistry. 1971 Jan 19;10(2):214-24
PMID: 5539223
-
LIPOPOLYSACCHARIDE OF THE GRAM-NEGATIVE CELL WALL.
Science. 1964 Aug 21;145(3634):783-9
PMID: 14163315
-
Composition and structure of the O-specific side chain of endotoxin from Serratia marcescens Bizio.
Biochemistry. 1973 Jan 16;12(2):309-15
PMID: 4345583
-
Degradation studies on the lipopolysaccharide from E. coli 071:K?:H12. Separation and investigation of O-specific and core polysaccharides.
FEBS Lett. 1968 Oct;1(5):311-314
PMID: 11945329
-
Molecular aspects of endotoxic reactions.
Bacteriol Rev. 1969 Mar;33(1):72-98
PMID: 4889150
-
Immunodiffusion studies on Escherichia coli. 1. Identification of O, K and H antigens in an O6 strain.
Acta Pathol Microbiol Scand. 1969;76(2):304-18
PMID: 4984753
-
Studies on the protein moiety of endotoxin from gram-negative bacteria. Characterization of the protein moiety isolated by phenol treatment of endotoxin from Serratia marcescens 08 and Escherichia coli 0 141:K85(B).
Eur J Biochem. 1971 Apr;19(3):340-56
PMID: 4928879
-
Chemical structure of lipid A of Selenomonas ruminantium.
J Biochem. 1971 Jul;70(1):187-91
PMID: 5562340
-
Evaluation of some extraction methods for the preparation of bacterial lipopolysaccharides for structural analysis.
Acta Pathol Microbiol Scand B Microbiol Immunol. 1972;80(5):751-9
PMID: 4629362
-
The linkage between the D-glucosamine units of lipid A isolated from the lipopolysaccharide of Serratia marcescens.
Biochim Biophys Acta. 1969 Oct 28;187(3):457-9
PMID: 4900193
-
Heterogeneity of endotoxic bacterial lipopolysaccharides revealed by ion-exchange column chromatography.
Nature. 1966 Apr 16;210(5033):278-80
PMID: 4959339
-
Structure of cell wall lipopolysaccharide from Salmonella typhimurium. Further studies on the linkage between O side chains and R core.
Eur J Biochem. 1970 Jul;15(1):57-62
PMID: 4923158
-
Studies on the characterization of lipopolysaccharides from two strains of Serratia marcescens.
Ann N Y Acad Sci. 1966 Jun 30;133(2):546-65
PMID: 4960346
-
Chemical and biological properties of an extracellular lipopolysaccharide from Escherichia coli grown under lysine-limiting conditions.
Biochem J. 1966 Apr;99(1):53-61
PMID: 5337757
-
The products of the partial acid hydrolysis of the mucopeptide from cell walls of Micrococcus lysodeikticus.
Biochem J. 1959 Aug;72:647-54
PMID: 14431858
-
Composition and structure of the O-specific side chain of endotoxin from Serratia marcescens 08.
Biochemistry. 1973 May 8;12(10):1948-55
PMID: 4350107
-
The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification.
J Biol Chem. 1959 Apr;234(4):705-9
PMID: 13654246
-
The determination of glucosamine and galactosamine.
Biochem J. 1955 Dec;61(4):586-9
PMID: 13276341
-
Evidence for two regions in the polysaccharide moiety of the lipopolysaccharide of Pseudomonas aeruginosa 8602.
FEBS Lett. 1970 Jul 29;9(2):81-84
PMID: 11947636
-
Studies in immunochemistry: The use of phenol and of alkali in the degradation of antigenic material isolated from Bact. dysenteriae (Shiga).
Biochem J. 1941 Nov;35(10-11):1140-63
PMID: 16747399