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.

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.

CONSTITUTIONS OF POLYSACCHARIDES FROM SERRATIA MARCESCENS

1962 ◽  
Vol 40 (7) ◽  
pp. 1415-1424 ◽  
Author(s):  
H. C. Srivastava ◽  
G. A. Adams
Keyword(s):  
Serratia Marcescens ◽  
Acid Hydrolysis ◽  
Main Chain ◽  
Degree Of Branching ◽  
Chemical Structures ◽  
Mannuronic Acid ◽  
Phenol Extraction ◽  
Glycosidic Bonds ◽  
Similar Chemical ◽  
Acidic Components

Constitutional studies of three polysaccharides prepared from Serratia marcescens cells by a sequence of phenol extraction, ultracentrifugation, and fractionation by Cetavlon are described. Methylation of the polysaccharides followed by acid hydrolysis yielded 2,3,4,6-tetra-O-methyl-D-glucose, 2,4,6-tri-O-methyl-D-glucose, 2,4,6-tri-O-methyl-D-mannose, 4,6-di-O-methyl-D-glucose, 2,6-di-O-methyl-D-glucose, and an unidentified di-O-methyl-glucose. The acidic components found were: 2,3,4-tri-O-methyl-D-mannuronic acid, 3-O-(2,3,4-tri-O-methyl-D-mannuronosyl)-2,4,6-tri-O-methyl-D-glucose, and O-2,3,4-tri-O-methyl-D-mannuronosyl-(1 → 3)-O-2,4,6-tri-O-methyl-D-glucosyl-(1 → 3)-2,4,G-tri-O-methyl-D-glucose. The polysaccharides are composed of a main chain of D-glucose and D-mannose residues joined by 1,3-glycosidic bonds. Some glucose residues carry branches at C2 and C4 which terminate in either D-glucose or D-mannuronic acid residues. The three polysaccharides studied had similar chemical structures but varied in the amounts of component sugars and degree of branching.

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.

POLYSACCHARIDE-PEPTIDE COMPLEXES FROM SERRATIA MARCESCENS CELLS

1964 ◽  
Vol 42 (5) ◽  
pp. 593-603 ◽  
Author(s):  
Hugh J. Creech ◽  
Evelyn R. Breuninger ◽  
G. A. Adams
Keyword(s):  
Chemical Composition ◽  
Serratia Marcescens ◽  
High Speed ◽  
Complete Removal ◽  
Complete Regression ◽  
Aqueous Extracts ◽  
Amino Acid Residues ◽  
Phenol Extraction ◽  
Peptide Complexes ◽  
Antitumor Properties

Determinations were made of the chemical composition and antitumor properties of several polysaccharide fractions obtained by high speed centrifugation of aqueous extracts of the cells of Serratia marcescens following phenol extraction and trypsin digestion. The use of methods that ensured complete removal of lipid gave polysaccharides containing firmly bound amino acid residues. These were highly effective in causing complete regression of well established solid tumors in mice. The products derived primarily from the cell wall were more potent than the polysaccharides of extracellular and cytoplasmic origins. Extensive differences were also noted in the chemical composition of the polysaccharide complexes isolated from the two sources.

URONIC ACIDS FROM WHITE SPRUCE (PICEA GLAUCA (MOENCH) VOSS)

1959 ◽  
Vol 37 (1) ◽  
pp. 29-34 ◽  
Author(s):  
G. A. Adams
Keyword(s):  
Acid Hydrolysis ◽  
Picea Glauca ◽  
Glucuronic Acid ◽  
White Spruce ◽  
Uronic Acids ◽  
Spruce Wood ◽  
Acidic Sugars ◽  
Hydrolysis Of ◽  
Acidic Components

Acid hydrolysis of extractive-free white spruce wood produced a number of neutral and acidic sugars and oligosaccharides. The acidic components were isolated and three of these were shown to be 4-O-methyl-D-glucuronic acid, 2-O-(4-O-methyl-α-D-glucopyranosyluronic acid)-D-xylose, and tentatively O-(4-O-methyl-α-D-glucopyranosyluronic acid)-(1→ 2)-O-β-D-xylopyranosyl-(1→ 4)-D-xylopyranose.

scholarly journals Identification and biochemical characterization of two novel UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucuronic acid 2-epimerases from respiratory pathogens

2007 ◽  
Vol 405 (1) ◽  
pp. 123-130 ◽  
Author(s):  
Erin L. Westman ◽  
David J. Mcnally ◽  
Martin Rejzek ◽  
Wayne L. Miller ◽  
Vellupillai Sri Kannathasan ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Glucuronic Acid ◽  
Biochemical Characterization ◽  
Intermediate Compound ◽  
Proton Spectrum ◽  
Respiratory Pathogens ◽  
Sodium Phosphate Buffer ◽  
Mannuronic Acid ◽  
Enzyme Substrate

The heteropolymeric O-antigen of the lipopolysaccharide from Pseudomonas aeruginosa serogroup O5 as well as the band-A trisaccharide from Bordetella pertussis contain the di-N-acetylated mannosaminuronic acid derivative, β-D-ManNAc3NAcA (2,3-diacetamido-2,3-dideoxy-β-D-mannuronic acid). The biosynthesis of the precursor for this sugar is proposed to require five steps, through which UDP-α-D-GlcNAc (UDP-N-acetyl-α-D-glucosamine) is converted via four steps into UDP-α-D-GlcNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucuronic acid), and this intermediate compound is then epimerized by WbpI (P. aeruginosa), or by its orthologue, WlbD (B. pertussis), to form UDP-α-D-ManNAc3NAcA (UDP-2,3-diacetamido-2,3-dideoxy-α-D-mannuronic acid). UDP-α-D-GlcNAc3NAcA, the proposed substrate for WbpI and WlbD, was obtained through chemical synthesis. His6–WbpI and His6–WlbD were overexpressed and then purified by affinity chromatography using FPLC. Capillary electrophoresis was used to analyse reactions with each enzyme, and revealed that both enzymes used UDP-α-D-GlcNAc3NAcA as a substrate, and reacted optimally in sodium phosphate buffer (pH 6.0). Neither enzyme utilized UDP-α-D-GlcNAc, UDP-α-D-GlcNAcA (UDP-2-acetamido-2,3-dideoxy-α-D-glucuronic acid) or UDP-α-D-GlcNAc3NAc (UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucose) as substrates. His6–WbpI or His6–WlbD reactions with UDP-α-D-GlcNAc3NAcA produce a novel peak with an identical retention time, as shown by capillary electrophoresis. To unambiguously characterize the reaction product, enzyme–substrate reactions were allowed to proceed directly in the NMR tube and conversion of substrate into product was monitored over time through the acquisition of a proton spectrum at regular intervals. Data collected from one- and two-dimensional NMR experiments showed that His6–WbpI catalysed the 2-epimerization of UDP-α-D-GlcNAc3NAcA, converting it into UDP-α-D-ManNAc3NAcA. Collectively, these results provide evidence that WbpI and WlbD are UDP-2,3-diacetamido-2,3-dideoxy-α-D-glucuronic acid 2-epimerases.

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