CELLULAR POLYSACCHARIDES OF SERRATIA MARCESCENS

1967 ◽  
Vol 45 (4) ◽  
pp. 477-491 ◽  
Author(s):  
G. A. Adams ◽  
S. M. Martin
Keyword(s):  
Cell Wall ◽  
Boric Acid ◽  
Serratia Marcescens ◽  
Glucuronic Acid ◽  
Analytical Data ◽  
Extracellular Polysaccharides ◽  
Similar Composition ◽  
Mannuronic Acid

Decapsulated cells of Serratia marcescens were fractionated to yield crude cytoplasmic (I) lipopolysaccharide (II) and cell-wall (III) polysaccharides. Further separation of I yielded dialyzable polysaccharides composed of D-glucose and D-mannose and nondialyzable polysaccharides containing various proportions of D-glucose, D-mannose, L-rhamnose, glucuronic acid, and glucosamine. Fractionation of II by precipitation with Cetavlon–boric acid and by highspeed centrifugation yielded polysaccharides containing D-glucose, L-rhamnose, D-mannose, D-glycero-D-manno-heptose, L-glycero-D-mannose-heptose, glucuronic acid, mannuronic acid, and glucosamine. Analytical data on the various fractions indicated that these polysaccharides included an acidic glucomannan, a rhamnoglucan, and a heptoglucan. Polysaccharides of similar composition have been found in S. marcescens capsular and extracellular polysaccharides in an earlier investigation. Removal of the mucopeptide material from the cell-wall III left an insoluble polysaccharide residue composed of glucose and glucosamine units.

PREPARATION AND PROPERTIES OF POLYSACCHARIDE–LIPID COMPLEXES FROM SERRATIA MARCESCENS

1962 ◽  
Vol 40 (7) ◽  
pp. 905-918 ◽  
Author(s):  
H. C. Srivastava ◽  
Evelyn Breuninger ◽  
Hugh J. Creech ◽  
G. A. Adams
Keyword(s):  
Serratia Marcescens ◽  
Glucuronic Acid ◽  
Antitumor Agent ◽  
Biological Properties ◽  
Culture Conditions ◽  
Aqueous Acetone ◽  
Uronic Acids ◽  
High Solubility ◽  
Mannuronic Acid ◽  
Phenol Extraction

Several polysaccharide fractions possessing pronounced antitumor activity were isolated from the polysaccharide–lipid–protein complex elaborated by Serratia marcescens. These fractions were obtained after phenol extraction of the cellular material, by ultracentrifugation and precipitation with ethanol and Cetavlon. They contained glucose, mannose, rhamnose, glucosamine, glucuronic acid, and mannuronic acid in different proportions. Each representative fraction caused regression of well-established solid tumors in mice but the dosage requirements varied considerably. One fraction containing glucose, mannose, and glucosamine, but no uronic acids, and having a high solubility in dilute sodium chloride solution and in aqueous acetone, was an exceptionally potent antitumor agent. Another fraction containing galactose and arabinose in addition to several of the usual components was found to be least active. From this work and that of other investigators, it is apparent that Serratia marcescens produces a spectrum of polysaccharides and lipopolysaccharides, the chemical composition and biological properties of which depend on the strain of organism, the culture conditions, and the methods employed for isolation of the active agents.

EXTRACELLULAR POLYSACCHARIDES OF SERRATIA MARCESCENS

1964 ◽  
Vol 42 (10) ◽  
pp. 1403-1413 ◽  
Author(s):  
G. A. Adams ◽  
S. M. Martin
Keyword(s):  
Serratia Marcescens ◽  
Culture Filtrate ◽  
Glucuronic Acid ◽  
Carbon Sources ◽  
Growth Period ◽  
The Other ◽  
Extracellular Polysaccharides ◽  
Active Growth ◽  
Electrophoretic Behavior ◽  
Active Growth Period

Growth of Serratia marcescens on sucrose, D-glucose, D-galactose, and D-xylose as carbon sources did not affect the composition of the extracellular polysaccharides significantly. D-Glucose was the major component with lesser amounts of D-mannose, heptose, L-fucose, and L-rhamnose. Rhamnose did not appear until near the end of the active growth period and increased proportionately more than the other sugars thereafter. From the culture filtrate after 20 hours growth on sucrose, two acidic polysaccharides were isolated. They were markedly different in composition and electrophoretic behavior although both contained glucose as their major component. One was characterized by a relatively high content of rhamnose and heptose, the other by the presence of mannose; both contained glucuronic acid. Other impure polysaccharides were isolated from the culture filtrate. It seems likely that S. marcescens produced a spectrum of rather similar extracellular polysaccharides of which the two isolated ones comprise the main types.

PREPARATION AND PROPERTIES OF POLYSACCHARIDE–LIPID COMPLEXES FROM SERRATIA MARCESCENS

1962 ◽  
Vol 40 (1) ◽  
pp. 905-918 ◽  
Author(s):  
H. C. Srivastava ◽  
Evelyn Breuninger ◽  
Hugh J. Creech ◽  
G. A. Adams
Keyword(s):  
Serratia Marcescens ◽  
Glucuronic Acid ◽  
Antitumor Agent ◽  
Biological Properties ◽  
Culture Conditions ◽  
Aqueous Acetone ◽  
Uronic Acids ◽  
High Solubility ◽  
Mannuronic Acid ◽  
Phenol Extraction

Several polysaccharide fractions possessing pronounced antitumor activity were isolated from the polysaccharide–lipid–protein complex elaborated by Serratia marcescens. These fractions were obtained after phenol extraction of the cellular material, by ultracentrifugation and precipitation with ethanol and Cetavlon. They contained glucose, mannose, rhamnose, glucosamine, glucuronic acid, and mannuronic acid in different proportions. Each representative fraction caused regression of well-established solid tumors in mice but the dosage requirements varied considerably. One fraction containing glucose, mannose, and glucosamine, but no uronic acids, and having a high solubility in dilute sodium chloride solution and in aqueous acetone, was an exceptionally potent antitumor agent. Another fraction containing galactose and arabinose in addition to several of the usual components was found to be least active. From this work and that of other investigators, it is apparent that Serratia marcescens produces a spectrum of polysaccharides and lipopolysaccharides, the chemical composition and biological properties of which depend on the strain of organism, the culture conditions, and the methods employed for isolation of the active agents.

CAPSULAR POLYSACCHARIDES OF SERRATIA MARCESCENS

1965 ◽  
Vol 43 (9) ◽  
pp. 1499-1512 ◽  
Author(s):  
G. A. Adams ◽  
Robert Young
Keyword(s):  
Serratia Marcescens ◽  
Culture Filtrate ◽  
Glucuronic Acid ◽  
Analytical Data ◽  
Capsular Polysaccharides ◽  
Carbohydrate Structure ◽  
Guluronic Acid ◽  
Electrophoretic Behavior ◽  
Sucrose Medium ◽  
Acidic Component

Four polysaccharides were isolated from the capsular layer of Serratia marcescens cells grown on a sucrose medium. In composition, electrophoretic behavior, and sedimentation in the ultracentrifuge, all were shown to be markedly different. Analytical data indicated that the polysaccharides were an acidic glucomannan (I), a rhamnoglucan (II), a glucoheptan (III), and a rhamnoheptoglucan (IV). In I, in addition to the main acidic component D-glucuronic acid, a small amount of guluronic acid was also identified. A relatively high protein or peptide content in the purified polysaccharides suggested that it was firmly bound to the carbohydrate structure; "bound" lipid consisting mainly of hydroxymyristic acid was also a part of the structure. Methylation studies established the main glycosidic linkages in I, II, and III. The capsular polysaccharides had properties similar to those of polysaccharides isolated from a culture filtrate of S. marcescens in an earlier study, and a possible relationship between these two groups of polysaccharides is discussed.

Automated Fluorometric Analysis of Calcium Pantothenate in Multivitamin Preparations

1978 ◽  
Vol 61 (3) ◽  
pp. 720-726
Author(s):  
Ram B Roy ◽  
Anthony Buccafuri
Keyword(s):  
Acid Solution ◽  
Boric Acid ◽  
Fluorescence Intensity ◽  
Analytical Data ◽  
Boric Acid Solution ◽  
Calcium Pantothenate ◽  
Magnesium Trisilicate ◽  
On Line ◽  
The Mean ◽  
Fluorometric Analysis

Abstract An automated fluorometric procedure is described for assaying calcium pantothenate in multivitamin preparations. Sample extracts containing calcium pantothenate are treated on-line with a slurry of magnesium trisilicate which removes any interfering riboflavin that may be present. The nitrate is resampled, mixed online with a slurry of Dowex 50W-X4 (H+) which removes any interfering β-alanine that may be present, and dialyzed. Dialysates are hydrolyzed in an alkaline medium and reacted with a mixture of o-phthalaldehyde and 2-mercaptoethanoI in boric acid solution. The fluorescence intensity due to the formation of a fluorogenic compound is measured at 455 nm after excitation at 350 nm. The procedure developed is capable of analyzing 20 samples/hr. Analytical data indicate that calcium pantothenate is assayed reliably both from real and synthetic multivitamin preparations. The mean recovery of calcium pantothenate added to sample solutions of tablet composites was 95.4%.

scholarly journals Capsule Production and Glucose Metabolism Dictate Fitness duringSerratia marcescensBacteremia

mBio ◽  
2017 ◽  
Vol 8 (3) ◽  
Author(s):  
Mark T. Anderson ◽  
Lindsay A. Mitchell ◽  
Lili Zhao ◽  
Harry L. T. Mobley
Keyword(s):  
Glucose Metabolism ◽  
Serratia Marcescens ◽  
Opportunistic Infections ◽  
Opportunistic Pathogen ◽  
Extracellular Polysaccharides ◽  
Mammalian Host ◽  
Wild Type ◽  
Transposon Insertion ◽  
Capsule Production ◽  
Tract Infections

ABSTRACTSerratia marcescensis an opportunistic pathogen that causes a range of human infections, including bacteremia, keratitis, wound infections, and urinary tract infections. Compared to other members of theEnterobacteriaceaefamily, the genetic factors that facilitateSerratiaproliferation within the mammalian host are less well defined. Anin vivoscreen of transposon insertion mutants identified 212S. marcescensfitness genes that contribute to bacterial survival in a murine model of bloodstream infection. Among those identified, 11 genes were located within an 18-gene cluster encoding predicted extracellular polysaccharide biosynthesis proteins. A mutation in thewzxgene contained within this locus conferred a loss of fitness in competition infections with the wild-type strain and a reduction in extracellular uronic acids correlating with capsule loss. A second gene,pgm, encoding a phosphoglucomutase exhibited similar capsule-deficient phenotypes, linking central glucose metabolism with capsule production and fitness ofSerratiaduring mammalian infection. Further evidence of the importance of central metabolism was obtained with apfkAglycolytic mutant that demonstrated reduced replication in human serum and during murine infection. An MgtB magnesium transporter homolog was also among the fitness factors identified, and anS. marcescens mgtBmutant exhibited decreased growth in defined medium containing low concentrations of magnesium and was outcompeted ~10-fold by wild-type bacteria in mice. Together, these newly identified genes provide a more complete understanding of the specific requirements forS. marcescenssurvival in the mammalian host and provide a framework for further investigation of the means by whichS. marcescenscauses opportunistic infections.IMPORTANCESerratia marcescensis a remarkably prolific organism that replicates in diverse environments, including as an opportunistic pathogen in human bacteremia. The genetic requirements forS. marcescenssurvival in the mammalian bloodstream were defined in this work by transposon insertion sequencing. In total, 212 genes that contribute to bacterial fitness were identified. When sorted via biological function, two of the major fitness categories identified herein were genes encoding capsule polysaccharide biogenesis functions and genes involved in glucose utilization. Further investigation determined that certain glucose metabolism fitness genes are also important for the generation of extracellular polysaccharides. Together, these results identify critical biological processes that allowS. marcescensto colonize the mammalian bloodstream.

scholarly journals Extracellular Polysaccharides from Xanthomonas axonopodis pv. manihotis Interact with Cassava Cell Walls During Pathogenesis

1997 ◽  
Vol 10 (7) ◽  
pp. 803-811 ◽  
Author(s):  
B. Boher ◽  
M. Nicole ◽  
M. Potin ◽  
J. P. Geiger
Keyword(s):  
Cell Wall ◽  
Monoclonal Antibodies ◽  
Host Cell ◽  
Cell Walls ◽  
Plant Cell Wall ◽  
Infection Process ◽  
Extracellular Polysaccharides ◽  
Xanthomonas Axonopodis ◽  
Cassava Leaves ◽  
Host Cell Wall

The location of lipopolysaccharides produced by Xanthomonas axonopodis pv. manihotis during pathogenesis on cassava (Manihot esculenta) was determined by fluorescence and electron microscopy immunolabeling with monoclonal antibodies. During the early stages of infection, pathogen lipopolysaccharides were detected on the outer surface of the bacterial envelope and in areas of the plant middle lamellae in the vicinity of the pathogen. Later in the infection process, lipopolysaccharide-specific antibodies bound to areas where the plant cell wall was heavily degraded. Lipopolysaccharides were not detected in the fibrillar matrix filling intercellular spaces of infected cassava leaves. Monoclonal antibodies specific for the exopolysaccharide xanthan side chain labeled the bacteria, the fibrillar matrix, and portions of the host cell wall. The association of Xanthomonas lipopolysaccharides with host cell walls during plant infection is consistent with a role of these bacterial extracellular polysaccharides in the infection process.

scholarly journals The composition of the cell wall of Fusicoccum amygdali

1971 ◽  
Vol 125 (2) ◽  
pp. 461-471 ◽  
Author(s):  
K. W. Buck ◽  
M. A. Obaidah
Keyword(s):  
Cell Wall ◽  
Sodium Hydroxide ◽  
Cell Walls ◽  
Digestive Enzymes ◽  
Helix Pomatia ◽  
Water Soluble ◽  
Extracellular Polysaccharides ◽  
Lytic Enzymes ◽  
Similar Nature ◽  
Dark Blue

1. The cell wall of Fusicoccum amygdali consisted of polysaccharides (85%), protein (4–6%), lipid (5%) and phosphorus (0.1%). 2. The main carbohydrate constituent was d-glucose; smaller amounts of d-glucosamine, d-galactose, d-mannose, l-rhamnose, xylose and arabinose were also identified, and 16 common amino acids were detected. 3. Chitin, which accounted for most of the cell-wall glucosamine, was isolated in an undegraded form by an enzymic method. Chitosan was not detected, but traces of glucosamine were found in alkali-soluble and water-soluble fractions. 4. Cell walls were stained dark blue by iodine and were attacked by α-amylase, with liberation of glucose, maltose and maltotriose, indicating the existence of chains of α-(1→4)-linked glucopyranose residues. 5. Glucose and gentiobiose were liberated from cell walls by the action of an exo-β-(1→3)-glucanase, giving evidence for both β-(1→3)- and β-(1→6)-glucopyranose linkages. 6. Incubation of cell walls with Helix pomatia digestive enzymes released glucose, N-acetyl-d-glucosamine and a non-diffusible fraction, containing most of the cell-wall galactose, mannose and rhamnose. Part of this fraction was released by incubating cell walls with Pronase; acid hydrolysis yielded galactose 6-phosphate and small amounts of mannose 6-phosphate and glucose 6-phosphate as well as other materials. Extracellular polysaccharides of a similar nature were isolated and may be formed by the action of lytic enzymes on the cell wall. 7. About 30% of the cell wall was resistant to the action of the H. pomatia digestive enzymes; the resistant fraction was shown to be a predominantly α-(1→3)-glucan. 8. Fractionation of the cell-wall complex with 1m-sodium hydroxide gave three principal glucan fractions: fraction BB had [α]D +236° (in 1m-sodium hydroxide) and showed two components on sedimentation analysis; fraction AA2 had [α]D −71° (in 1m-sodium hydroxide) and contained predominantly β-linkages; fraction AA1 had [α]D +40° (in 1m-sodium hydroxide) and may contain both α- and β-linkages.

THE POLYSACCHARIDES OF CRYPTOCOCCUS LAURENTII (NRRL Y-1401): PART I

1960 ◽  
Vol 38 (9) ◽  
pp. 1617-1624 ◽  
Author(s):  
M. J. Abercrombie ◽  
J. K. N. Jones ◽  
M. V. Lock ◽  
M. B. Perry ◽  
R. J. Stoodley
Keyword(s):  
Glucuronic Acid ◽  
Extracellular Polysaccharides ◽  
Cryptococcus Laurentii ◽  
Structural Studies ◽  
Acidic Polysaccharide ◽  
End Groups

The extracellular polysaccharides produced by Cryptococcuslaurentii have been isolated and shown to consist of (A) an acidic polysaccharide containing D-mannose, D-xylose, and D-glucuronic acid; (B) a neutral polysaccharide containing D-glucose only.Preliminary structural studies on the acidic material suggest that it consists of a mannose-containing backbone with xylose and glucuronic acid as end groups, while the glucan contains 1 → 3, 1 → 4, 1 → 2, and (or) 1 → 6 linked residues.

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