Polysaccharide fractionation

In the late 1940s Stacey, with the able and enthusiastic assistance of Paul Kent, examined polysaccharide material from Mycobacterium tuberculosis human strain. From heat-killed cells, two stable, serologically specific polysaccharide fractions and a degraded bacterial glycogen were isolated and examined. 

During the past decade, MALDI-TOF MS has proven to be an effective tool for the analysis of oligo- and polymeric mannoglucans (for extensive reviews see [222,223]). SEC/MALDI mass spectrometry was employed in the analysis of hemicelluloses isolated by microwave heat-fractionation from spruce and aspen wood [94]. These methods allowed the separation and characterization of the oligo- and polysaccharide fractions derived from the xylan and mannan components of both woods [224]. 

Ginsenan S-IIA, a polysaccharide fraction from the roots of P. ginseng is a potent inducer of IL-8 production by human monocytes and THP-1 cells, and this induction is accompanied by increased IL-8 mRNA expression. The polysaccharide appears from the structural feature to be a mixture of arabino-galactan type I and type II, based on the presence of 1,3-, 1,6-, 1,3,6-, 1,4-, and 1,4,6-galactose units as well as terminal arabinose and 1,5-, 1,3,5-, and 1,2,5-linked units. It also contains 1,4,6-linked glucose units that together with the 1,2,5-linked arabinose units are different from the units found in other ginseng polysaccharides and may thus be of importance for the activity [64]. 

Already in 1988 and 1991, Gao et al. [65,66] detected four different polysaccharides present in the leaves of Panax ginseng that had an effect on the complement system, but only two of them, the neutral, GL-NIa, and one of the acidic ones, GL-AIa, had potent activities at low concentrations. GL-NIa was found to be mainly an arabinigalactan type II polymer. GL-AIa was a polysaccharide with a rhamnogalacturonan core with neutral side chains of the AG-II type, confirmed by a strong reaction with the Yariv reagent and the methylation results. It was shown that the crude polysaccharide fraction contained KDO and DHA, suggesting the presence of Rhamnogalacturonan II in... 

Four different European herbaceous plants have also been tested for possible mitogenic and comitogenic activities. Using Zymosan as a positive control, all polysaccharide fractions tested had higher activity in the bioassays... 

A cell-wall polysaccharide fraction remaining after treatment with a "classical" pectinase is termed rhamnogalacturonan (RG) or modified hairy regions (MHR). It is characterized by a highly branched ihamnogalacturonan-polymer with some arabinan side chains. 

Chromatorapf c methods For gel filtration of polysaccharide fraction PI, a Sephacryl S-300 chromatographyc column (1,1 X 46,7 cm) was calibrated with standard dextrans (molecular mass range 266, 72, 40, and 17 KDa Sigma Chemicals), and the void volume determined with blue dextran. Polysaccharide sample (0.5 mL 2 mg/mL) was applied and eluted with 50 mM NaOH, fractions 1 mL being collected and carbohydrate absorbance (phenol-H2S04) being monitored. 

Recently, We have found that the crude polysaccharide fraction (GL-2) from the leaves and the purified polysaccharide (GL-4IIb2 ) from GL-2, showed potent... 

Crude polysaccharide fraction (GL-2) was prepared from the leaves of P. ginseng by hot water extraction, ethanol precipitation and dialysis, and GL-2 was fractionated by Cetavlon precipitation and weakly acidic polysaccharide fraction (GL-4) was obtained[3]. GL-4IIb2 was purified from GL-4 by DEAE-Sepharose CL-6B as described previousely [3]. In order to remove the color-materials, GL-4IIb2 was further purified by Q-Sepharose (C1 form), and the major fraction, eluted with 0.3 M NaCl, was repurifled by gel filtration on Bio-gel P-30 column to obtain purified active polysaccharide, GL-4IIb2. ... 

Isolation of polysaccharide fractions After a growth period of 8 days the cell biomass was separated by filtration and the spent culture medium was used for the isolation of polysaccharides. The following procedures were examined ... 

Gel chromatography on Sephadex GlOO (2.8x50cm) of polysaccharide fraction I (Sample 2). The polysaccharide fraction was dissolved in a phosphate buffer at pH 6. After centrifugation, the supernatant was applied to the column at a flow rate of 0.8 ml/min. The elution was performed with a phosphate buffer and fractions of 10 ml each were collected. The refraction of each fraction was measured interferometrically. Fractions with coincident peaks were collected and analyzed for content of galacrturonic acid, neutral sugars and protein. 

Determination of number o/per/ro/jenZ-exudative cells after i.p. (intraperitoneal) application of polysaccharide fractions and during infection with Y. pseudotuberculosis in experimental animals. 

Bioassay for "killing" ability (in vivo and in vitro against bacteria) of peritonial macrophages after treatment with polysaccharide fractions. 

The uronic acids in polysaccharide fraction I (Table 2) were determined colorimetrically with m-hydroxybiphenyl (15). 

The protein content of the polysaccharide fractions was determined by the method of... 

A new possibility for isolation of the exopolysaccharides in deep freezing of the culture medium (-18°C) was arrived at in the course of our research (Sample 2, Polysaccharide fraction I). 

A second polysaccharide fraction (Sample 3) was isolated from the filtrates by concentration and following precipitation with ethanol (1 3). The yield and characteristics of the obtained polysaccharides (Samples 2 and 3) are given in Table 1. It is evident that the sum of the yields for Samples 2 and 3 is almost equal to the yield for Sample 1. The polyuronic content data are also well balanced. This fact indicates that the suggested method is suited for fractional isolation of the polysaccharides from the spent culture medium of H.annuus 1805 cell suspension. As can be seen from Table 1, the main part of the exopolysaccharide was in fraction I. 

The results of the biological investigation showed that polysaccharide fraction I had a distinct immunostimulating activity. 

Composition of polysaccharide fraction I, isolated from the culture medium of H. amuus 1805... 

The results suggest that polysaccharide fraction I may be thought as unspecific modulators of immune responsiveness. 

A structural study on lipid A and the O-specific polysaccharide of the lipopoly-saccharide from a clinical isolate of Bacteroides vulgatus from a patient with Crohn s disease was conducted by Hashimoto and coworkers [39]. They separated two potent virulence factors, capsular polysaccharide (CPS) and lipopolysaccharide (LPS), from a clinical isolate of B. vulgatus and characterized the structure of CPS. Next, they elucidated the strucmres of O-antigen polysaccharide (OPS) and lipid A in the LPS. LPS was subjected to weak acid hydrolysis to produce the lipid A fraction and polysaccharide fraction. Lipid A was isolated by PLC, and its structure was determined by MS and NMR. 

Campbell, D.H. (1936) An antigenic polysaccharide fraction of Ascaris lumbricoides (from hog). Journal of Infectious Diseases 59, 266-280. 

Carcinogenesis inhibition. Polysaccharide fraction of the dried seed hulls, administered to rats by gastric intubation, was active vs tumor induction with N-ethyl N-nitro-N nitrosoguanidine h Rice bran, administered orally to male rats at a concentration of 4% of the diet, was active. A 1 1 combination of wheat bran and psyllium, at a total level of 8% dietary fiber, offers the highest protection against colon tumor development . 

Glutamate-oxaloacetate-transaminase stimulation. Polysaccharide fraction of the dried stem, administered intragastrically to rats at a dose of 1 g/kg, was active vs high-sugar dieh° h... 

Hypoglycemic activity. Ethanol (95%) extract of the dried leaf, administered intragastrically to rabbits at a dose of 1 g/kg, produced weak activity ". Juice of the dried stalk, administered intraperitoneally to mice at a dose of 200 mg/kg, was active . Polysaccharide fraction of the dried stem, administered intraperitoneally to rats at a dose of 40 mg/kg, was inactive vs high-sugar diet °" . 

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