SUMMARY: Two mutants which lacked both capsular and lipopolysaccharide O-antigen polysaccharides were isolated from Klebsiella aerogenes serotype 2 by phage selection; these were designated rough mutants. The polysaccharide fractions solubilized by partial acid hydrolysis of the lipopolysaccharide from both the wild type and mutants were chromatographed on Sephadex G-50. Analysis of the fractions obtained confirmed that the rough mutants lacked the galactan portion of the molecule, which is analogous to the Salmonella O-antigen polysaccharide.
Membranes prepared from wild-type K. aerogenes, from a non-mucoid strain (lacking capsule only), and from one of the rough mutants were used in incubation mixtures to compare the biosynthesis of polysaccharides by these organisms. The incorporation of sugar nucleotides into both lipid intermediates and polymer was followed. Results show that the transferases were apparently present in all membranes, while the polymerases were absent in both the non-mucoid and rough mutants.
AndersonJ. S.,
MatsuhasiM.,
HaskinM.,
StromingerJ. L.1965; Lipid-phosphoacetylmuramyl-pentapeptide and lipid-phospho-disaccharide-pentapeptide: presumed membrane transport intermediates in cell-wall synthesis. Proceedings of the National Academy of Sciences of the United States of America 53:881–889
DuboisM.,
GillesK. A.,
HamiltonJ. K.,
RebersP. A.,
SmithF.1956; Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28:350–356
FensomA. H.,
MeadowP. M.1970; Evidence for two regions in the polysaccharide moiety of the lipolysaccharide of Pseudomonas aeruginosa8602. FEBS Letter 9:81–84
GahanL. C.,
SandfordP. A.,
ConradH. E.1967; The structure of the serotype 2 capsular polysaccharide of Aerobacter aerogenes. Biochemistry; New York: 62755–2767
KoeltzowD. E.,
EpleyJ. D.,
ConradH. E.1968; The lipopolysaccharides of Aerobacter aerogenes strains A3 (s1) and NCTC243. Biochemistry; New York: 72920–2928
LowryO. H.,
RosebroughN. J.,
FarrA. L.,
RandallR. J.1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
NikaidoH.1973; Biosynthesis and assembly of lipopolysaccharides and the outer membrane layer of Gram-negative cell wall. In Bacterial Membranes and Walls pp. 131–208LieveL.
Edited by New York:: Dekker.;
NikaidoH.,
NikaidoK.,
NakaeT.1971; Glucosylation of lipopolysaccharide in Salmonella: Biosynthesis of O antigen factor 122. Journal of Biological Chemistry 246:3902–3911
OsbornM. J.1963; Studies on the Gram-negative cell wall. I. Evidence for the role of 2-keto-3-deoxy-octonate in the lipopolysaccharide of Salmonella typhimurium. Proceedings of the National Academy of Sciences of the United States of America 50:499–506
SchultzJ.,
ElbeinA. D.1974; Biosynthesis of mannosyl- and glucosyl-phosphoryl polyprenols in Mycobacterium smegmatis. Archives of Biochemistry and Biophysics 160:311–322
StromingerJ. L.,
ParkJ. T.,
ThompsonR. E.1959; Composition of the cell wall of Staphylococcus aureus: its relation to the mechanism of action of penicillin. Journal of Biological Chemistry 234:3263–3268
SutherlandI. W.,
NorvalM.1970; The synthesis of exopolysaccharides by Klebsiella aerogenes membrane preparations and the involvement of lipid intermediates. Biochemical Journal 120:567–576
SutherlandI. W.,
WilkinsonJ. F.1965; Depolymerases for bacterial polysaccharides obtained from phage-infected bacteria. Journal of General Microbiology 39:373–383
SutherlandI. W.,
WilkinsonJ. F.1966; The composition of lipopolysaccharides of Klebsiella aerogenes and Aerobacter cloacae. Biochimica et biophysica acta 117:261–263