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PubMed2000

Control of Mung bean pectinmethylesterase isoform activities. Influence of pH and carboxyl group distribution along the pectic chains.

Well-characterized pectin samples with a wide range of degrees of esterification (39-74%) were incubated with the solubilized pure alpha and gamma isoforms of pectinmethylesterase, from mung bean hypocotyl (Vigna radiata). Enzyme activity was determined at regular intervals along the deesterification pathway at pH 5.6 and pH 7.6. It has been demonstrated that the distribution of the carboxyl units along the pectin backbone controls the activity of the cell wall pectinmethylesterases to a much greater extent than the methylation degree, with a random distribution leading to the strongest activity. Polygalacturonic acid was shown to be a competitive inhibitor of the alpha isoform activity at pH 5.6 and to inhibit the gamma isoform activity at both pH 5.6 and pH 7.6. Under these conditions, the drop in enzyme activity was shown to be correlated to the formation of deesterified blocks of 19 +/- 1 galacturonic acid residues through simulations of the enzymatic digestion according to the mechanisms established previously (Catoire, L., Pierron, M., Morvan, C., Herve du Penhoat, C., and Goldberg, R. (1998) J. Biol. Chem. 273, 33150-33156). However, even in the absence of inhibition by the reaction product, activity dropped to negligible levels long before the substrate had been totally deesterified. Comparison of alpha and gamma isoform cDNAs suggests that the N-terminal region of catalytic domains might explain their subtle differences in activity revealed in this study. The role of pectinmethylesterase in the cell wall stiffening process along the growth gradient is discussed.

Amino Acid Sequence↗

PubMed2003

Recovery of development and functionality of nodules and plant growth in salt-stressed Pisum sativum--Rhizobium leguminosarum symbiosis by boron and calcium.

Nodules developed in Pisum sativum L. cv. Argona inoculated with Rhizobium leguminosarum bv. viciae 3841 and growing under saline conditions (75 mmol/L NaCl) are non functional and had abnormal structure. The infected cells contained a low amount of endophytic bacteria, compared to treatments without salt. Addition of B (up to 55.8 micromol/L) and Ca2+ (up to 2.72 mmol/L) increased bacterial population of host plant cells in salt-stressed nodules. Furthermore, symbiosomes developed inside the nodules from salt treated plants presented a degraded peribacteroid membrane. This effect was also prevented by combined addition of B and Ca2+. Given the importance of both nutrients in cell wall structure, the pectin fraction was studied by electron microscopy and immunological methods. Salt stress produced cells with walls dramatically altered or even degraded in several zones. Pectin polysaccharides, detected by JIM 5 monoclonal antibody, increased in cells under salinity. These effects resembled typical effects of B-deficiency reactions in cell walls, and the increase of both Ca2+ and especially B also prevented these alterations.

Boron↗

PubMed1989

Intestinal zinc transfer by everted gut sacs from rats given diets containing different amounts and types of dietary fibre.

Two experiments were carried out in which rats were offered diets containing different amounts and types of dietary fibre, i.e. commercial stock diet and three semi-purified diets containing no fibre, 200 g wheat bran or 200 g pectin/kg. Dietary inclusion of fibre, and especially pectin, stimulated large bowel fermentation, as indicated by caecal hypertrophy and reduced caecal pH. After 3 weeks, mucosal:serosal zinc transfer and Zn accumulation by tissue were measured using the everted-gut-sac technique. In Expt 2, incubations were carried out in the presence and absence of 0.25 mM-ouabain to assess the importance of transfer by Na+,K+-ATPase-dependent mechanisms, and some observations on glucose transport were also made. Ouabain reduced rates of transfer of both Zn and glucose and also tissue Zn accumulation. There were no significant differences in rates of Zn transfer by everted sacs from duodenal, ileal and colonic sites, but accumulation of Zn by tissue was a more important fate than transfer across the serosal surface, and accumulation by duodenal tissue was approximately twice as great as by other tissues. Mucosal:serosal transfer of glucose by ileal tissue was much more sensitive to ouabain than was Zn transfer. Previous diet appeared to alter the capacity of the intestinal tissue to transfer Zn, with the highest rates of transfer being by colonic tissue from pectin-fed rats.

Animals↗

PubMed1992

Enzyme supplementation, degradation and metabolism of three U-14C-labelled cell-wall substrates in the fowl.

An experimental model is described that was used for assessing in vivo effects in fowls of exogenous enzyme supplementation on the degradation of plant cell walls to metabolizable monosaccharide residues. It was based on tube-feeding U-14C-labelled cell-wall substrates, cellulose, spinach (Spinacia oleracea) or Festuca, with and without enzyme treatments, and monitoring recovery of 14C radioactivity in exhaled carbon dioxide and excreta in the following 8 h. Normal digestion of cell-wall polysaccharides by endogenous microbial activity was also studied by pretreating birds with an antibiotic mixture intended to deplete their intestinal microflora. The results of this pretreatment appeared to confirm the existence of microbial degradation of cellulose in (conditioned) fowls. Judging from differences in 14CO2 production, effects of exogenous enzyme additions were greatly enhanced with all substrates by combining them with a wet pretreatment, thereby increasing the time-period available for them to act in aqueous conditions. However, estimations of digestibilities of cellulose, hemicellulose and pectin with dry and wet treatments, based on recovery of 14C in excreta, indicated that it was only cellulose digestion that was improved by the wet pretreatment. This suggests that degradation of cellulose, which appeared to be slowest, was limited by the dry treatments, whereas that of hemicellulose and pectin was not. Respective digestibilities of these three cell-wall components, from all treatments combined, were in the proportions 1:1.5:4.2.

Animals↗

PubMed1994

Dietary fibers stimulate colonic cell proliferation by different mechanisms at different sites.

The effect of dietary fiber type (cellulose, pectin, or oat bran) on colonic short-chain fatty acids (SCFA), luminal pH, and epithelial cell proliferation was examined in 60 male Sprague-Dawley rats. In vivo cell proliferation was measured by [3H]thymidine incorporation into DNA, pH was measured in vivo, and luminal SCFA were determined by gas chromatography. The pectin and oat bran diets produced higher concentrations of all types of SCFA and total SCFA than cellulose at every site. The higher the SCFA concentration, the lower was the pH at each site. In the cecum, the concentration of SCFA was positively associated with the number of cells per crypt column (r = 0.961), total cells per crypt (r = 0.963), and the proliferative zone (r = 0.845). In contrast, in the distal colon, there was no significant correlation between SCFA concentration and measurements of cell proliferation. These data suggest that fiber stimulates cecal cell proliferation through the production of SCFA. The effects of fiber on distal colonic cytokinetics are likely mediated through a different mechanism.

Animals↗

PubMedDate not supplied

Polymers for sustained release formulations of dipyridamole-alginate microspheres and tabletted microspheres.

The preparation of dipyridamole (DIP) alginate (alg) microspheres by different methods or the incorporation of tragacanth (trgh), pectin or Eudragit L-100 55 (Eud) in alg microsphere formulations did not provide a prolonged release of DIP at pH 1.2. Tabletted microsphere formulations containing alg, trgh, pectin, sodium carboxymethyl cellulose (CMC), sodium starch glycolate (SSG), carrageenan (carrg) or Eud as diluents in different ratios, produced tablets with good physical properties which did prolong DIP release. The type, viscosity and the ratio of the diluent polymer, microsphere size and the compression pressure were found to be important factors to produce tablets with desired properties. No advantage of the tablets containing alg microspheres and granulated diluents was observed over the tablets containing powdered diluents.

Alginates↗

PubMed1993

Preparation of a dietary fiber mixture derived from different sources and its metabolic effects in rats.

In this study we analyzed a commercial dietary fiber blend known as Fiber-Plus and a dietary fiber mixture (DFM) prepared in our lab (10% apple pectin, 10% orange pectin, 36% locust bean gum and 44% corncob composed of 56% fiber). DFM provided greater inhibition of starch digestion and glucose absorption (p < 0.05) than did Fiber-Plus. DFM was also found to be significantly more efficient (p < 0.05) than Fiber-Plus at reducing the glycemic response following starch loading. Rats fed DFM or corncob demonstrated a much lower glycemic response following a test meal as compared to those fed a fiber-free diet. Both fiber sources decreased fasting cholesterol and triglyceride concentrations; transit time was shortened and fecal values were higher than those of controls. beta-glucuronidase activity in intestinal segment contents and feces was lower in rats fed fiber. We conclude that DFM is effective in controlling plasma glucose and lipid levels. Corncob, a novel fiber source, could be beneficial in the treatment of gastrointestinal tract disorders and in the elimination of carcinogenic elements.

Animals↗

PubMed1978

Fiber, intestinal sterols, and colon cancer.

It has been postulated that dietary fiber's protective effect against the development of colon cancer, diverticular disease, and atherosclerosis may be due to the adsorption and/or dilution of intestinal sterols such as bile acids and neural sterols and their bacterial metabolites by component(s) of fiber. Dietary fiber is made up of four major components-cellulose, hemicellulose, lignin, and pectin. There is evidence that hemicellulose and pectin may induce an increase in fecal bile acid excretion in man which may be accompanied by a decrease in serum cholesterol. Natural fibers, such as rolled oats, alfalfa, guar gum, and Bengal gram have been shown to have hypocholesterolemic properties of alfalfa, wheat straw, and some other fibers found considerable amounts of bile acids in vitro. On the other hand, wheat bran, oat hulls, and all the synthetic fibers tested bound only negligible amounts of bile acids under the same conditions. Vegetarians in the United States have lower plasma lipids and different plasma lipoprotein patterns than those of comparable control populations on regular mixed diet. They also have smaller daily fractional turnover rates of cholic acid and deoxycholic acid pool size. In addition, populations on a mixed Western diet, where the rate of large bowel cancer is high (North American, English, Scottish, etc.) degraded and excreted cholesterol and bile acid metabolites to a greater degree than populations where the rate of colon cancer is comparatively low (Ugandan, Japanese, etc). It cannot be denied that the fiber theory linking fiber deficiency with the development of colon cancer and other diseases, is simple, attractive and appears to be firmly based in common sense. When subjected to research studies, however, the situation appears much more complex than expected. Although some progress is being made, the data are often contradictory and confusing, probably due to lack of adequate documentation of fiber intake (e.g., use of dietary fiber instead of crude fiber) and/or the absence of detailed information on the chemistry of the fiber itself.

Adult↗

PubMed1979

Hypocholesterolemic action of dietary fiber unrelated to fecal bulking effect.

Twenty-two healthy volunteers took approximately 20 g/day of concentrated dietary fiber from either carrot, cabbage, apple, bran, or guar gum or 31 g from pectin, added for 3-week periods to controlled diets. Total serum cholesterol fell by 13% on both guar and pectin (P less than 0.01) with no significant change in high density lipoprotein cholesterol. Over the 3-week supplementation period, the other fibers were without effect with the exception of carrot, where both control and test high density lipoprotein levels fell (P less than 0.05 and less than 0.01, respectively). If, however, the 3rd week of the control was compared with the 3rd test week, the values for total cholesterol were 7% lower after apple (P less than 0.02) while after carrot the high density lipoprotein cholesterol level was 10% lower than the control (P less than 0.01). No significant change was seen in serum triglyceride or body weight either as judged by differences over the 3-week periods or by comparing test and control values at 3 weeks. Comparison of stool weights obtained in this study indicate that the fecal bulking action of dietary fiber is independent of its hypocholesterolaemic effect.

Adult↗

PubMed1979

The effect of dietary fiber on postprandial serum digoxin concentration in man.

The postprandial digoxin concentrations in serum were measured for 4 hr by radioimmunological assay in six healthy volunteers who had received 0.8 mg beta-acetyldigoxin by the oral route. Digoxin was given together with a formula diet containing as admixture, respectively, wheat bran, microcrystalline cellulose, pectin, carrageenan, and carob seed flour. The various additions did not reduce the mean digoxin concentrations in serum. After carob seed flour, the concentrations were significantly (P less than 5%) higher, as compared to the control. After microcrystalline cellulose, pectin, and wheat bran the mean peak concentrations were reached later than in the control state.

Acetyldigoxins↗

PubMed1981

Effect of chronic intake of dietary fibers on the ultrastructural topography of rat jejunum and colon: a scanning electron microscopy study.

This report is an attempt to quantitate the observable topographical characteristics of small and large intestine after a specific dietary regimen under well-defined states of lipid absorption and metabolism. Alfalfa, white wheat bran, cellulose, and pectin were fed for 6 wk at a level of 15 g/100 g diet to four groups of rats (12 rats per dietary group). A 5th control group was maintained on Purina Rat Chow. Three animals from each group were blind-coded for morphological assessment. After anesthesia, the jejunum and mid-colon were removed and processed for scanning electron microscopy. Beginning with the mildest mucosal surface changes, the observed order in terms of increasing severity is bran less than cellulose less than pectin less than alfalfa. Our observations suggest that altered rates of cell loss in intestinal tract cytokinetics may be occurring with particular feeding patterns and should be considered as a possible mechanism in the nutritional consequences of dietary fiber intake.

Animals↗

PubMed1982

Components of fiber bind iron in vitro.

Interactions among fiber, inorganic iron, and substances known to chelate iron were examined in vitro. Cellulose, neutral and acid fractions of lignin, psyllium mucilage, and pectin were incubated with 59Fe SO4 at various pH's, in various concentrations, and in the presence or absence of known chelators of iron including ascorbate, citrate, cysteine, fructose, and EDTA. Insoluble components of fiber were tested in a precipitation-centrifugation system, soluble components were tested by equilibrium dialysis. Lignins and psyllium mucilage had the greatest capacity to bind ferrous iron under conditions that approximated those of the proximal intestine postprandially; cellulose and pectin were less potent. Dissociation constants of binding were similar for several components, suggesting the existence of comparable binding sites in different components of fiber. Citrate and EDTA inhibited the binding of iron markedly but other chelators were much less effective. These reactions among fiber, inorganic iron, and other constituents of food may influence the bioavailability of dietary iron and the simple systems described here offer a means of dissecting interactions that are complex in vivo.

Cellulose↗

PubMed1990

Cholesterol-lowering effects of soluble-fiber cereals as part of a prudent diet for patients with mild to moderate hypercholesterolemia.

Soluble-fiber breakfast cereals were examined for their cholesterol-lowering ability in 58 male patients with mild to moderate hypercholesterolemia in a randomized, double-blind, placebo-controlled study. Patients followed a step 1 diet for a minimum of 6 wk, then were randomly assigned to groups incorporating either corn flakes or one of two soluble-fiber cereals (pectin enriched or psyllium enriched) in the diet for an additional 6 wk. During the diet-only phase, total cholesterol dropped 3.8%. During the cereal-plus-diet phase, total and LDL cholesterol values of the pectin-enriched cereal group dropped an additional 2.1% (P = 0.243) and 3.9% (P = 0.16), respectively, and they dropped 5.9% (P = 0.005) and 5.7% (P = 0.034), respectively, in the psyllium-enriched cereal group. During the cereal-plus-diet phase, no significant effects on HDL cholesterol, triglyceride, or body weight were found within or between any cereal groups. These results support use of soluble-fiber cereals as an effective and well-tolerated part of a prudent diet in the treatment of mild to moderate hypercholesterolemia.

Adult↗

PubMed2005

Widespread occurrence of a covalent linkage between xyloglucan and acidic polysaccharides in suspension-cultured angiosperm cells.

BACKGROUND AND AIMS: Covalent linkages between xyloglucan and rhamnogalacturonan-I (RG-I) have been reported in the primary cell walls of cultured Rosa cells and may contribute to wall architecture. This study investigated whether this chemical feature is general to angiosperms or whether Rosa is unusual. * METHODS: Xyloglucan was alkali-extracted from the walls of l-[1-3H]arabinose-fed suspension-cultured cells of Arabidopsis, sycamore, rose, tomato, spinach, maize and barley. The polysaccharide was precipitated with 50 % ethanol and subjected to anion-exchange chromatography in 8 m urea. Eluted fractions were Driselase-digested, yielding [3H]isoprimeverose (diagnostic of [3H]xyloglucan). The Arabidopsis cells were also fed [6-14C]glucuronic acid, and radiolabelled pectins were extracted with ammonium oxalate. * KEY RESULTS: [3H]Xyloglucan was detected in acidic (galacturonate-containing) as well as non-anionic polysaccharide fractions. The proportion of the [3H]isoprimeverose units that were in anionic fractions was: Arabidopsis, 45 %; sycamore, 60 %; rose, 44 %; tomato, 75 %; spinach, 70 %; maize, 50 %; barley, 70 %. In Arabidopsis cultures fed d-[6-(14)C]glucuronate, 20 % of the (galacturonate-14C)-labelled pectins were found to hydrogen-bond to cellulose, a characteristic normally restricted to hemicelluloses such as xyloglucan. * CONCLUSIONS: Alkali-stable, anionic complexes of xyloglucan (reported in the case of Rosa to be xyloglucan-RG-I covalent complexes) are widespread in the cell walls of angiosperms, including gramineous monocots.

Cell Wall↗

PubMed2005

Resolution of the structural isomers of partially methylesterified oligogalacturonides by polysaccharide analysis using carbohydrate gel electrophoresis.

Pectins differing in their degree and pattern of methylesterification are important in diverse aspects of plant physiology and also in many industrial applications. Determination of methylesterification fine structure and knowledge of enzyme specificities in modification and fragmentation of pectin are key to understanding the relationship between structure and function. The development of methodologies for the detection, separation and sequencing of different partially methylesterified oligogalacturonides (Me-OGAs) is consequently very important. Polysaccharide analysis using carbohydrate gel electrophoresis (PACE) has been shown to be powerful for the quantitative resolution of species different in degree of polymerization (DP) and/or degree of methylesterification (DM). Mass spectrometry (MS) has, to date, been the only tool with which to obtain isomeric information. However, it is not quantitative, and the presence of isobaric species makes the interpretation of the fragmentation patterns complicated. Here, we present evidence that Me-OGAs with the same DP and DM but different patterns of methylesterification (structural isomers) can easily be separated and quantified using PACE.

Carbohydrate Conformation↗

PubMed2002

Guard cell wall: immunocytochemical detection of polysaccharide components.

The composition of guard cell walls in sugar beet leaves (Beta vulgaris L.) was studied by using histochemical staining and immunocytochemical detection of cell wall antigens. The findings were compared with those in the walls of epidermal and mesophyll cells. Probing of leaf sections with monoclonal antibodies against pectins, terminal fucosyl residues linked alpha-(1-->2) to galactose, beta-(1-->3)-glucans and arabinogalactan-proteins revealed several specific features of guard cells. Pectic epitopes recognized by JIM7 were homogeneously distributed in the wall, whereas pectins recognized by JIM5 were not found in the walls themselves, but were abundant in the cuticular layer. Large amounts of molecules bearing terminal fucose were located predominantly in ventral and lateral guard cell walls. Much smaller amounts were detected in dorsal walls of these cells, as well as in the walls of pavement and mesophyll cells. Conspicuous accumulation of these compounds was observed in the vicinity of the guard cell plasmalemma, whereas labelling was scarce in the areas of the wall adjacent to the cell surface. The presence of callose clearly marked the ventral wall between the recently formed, very young guard cells. Callose also appeared in some mature walls, where it was seen as punctate deposits that probably reflected a specific physiological state of the guard cells. Large amounts of arabinogalactan-proteins were deposited within the cuticle, and smaller amounts of these proteoglycans were also detected in other tissues of the leaf. The histochemical and immunocytochemical structure of the guard cell wall is discussed in the light of its multiple functions, most of which involve changes in cell size and shape.

Antibodies, Monoclonal↗

PubMed1982

Effects of dietary fiber on the bioavailability of folic acid monoglutamate.

Low bioavailability of folacin has been previously reported for a variety of foods of plant origin. This study was conducted to examine the possible role of various types of dietary fiber on the bioavailability of folic acid monoglutamate. Cellulose, pectin, lignin, sodium alginate and wheat bran were selected for their differing physical and chemical properties. In vitro binding studies by equilibrium dialysis showed no evidence of physical or chemical binding of folic acid under physiological conditions. In vivo effects were evaluated by a chick bioassay with graded levels of folic acid in semipurified diets containing the fiber materials at 3% (weight/weight). Total liver and plasma folacin concentration and chick growth were used as response indicators. Dose-response curves indicated that pectin, lignin and alginate significantly reduced chick growth at all levels of dietary folacin. Plasma and liver folacin dose-response curves were not significantly different for any of the fiber materials, which indicated that the growth impairment was not due to a fiber effect on folic acid absorption. These results suggest that added dietary fiber has little or no effect on the bioavailability of folic acid monoglutamate.

Alginates↗

PubMed1982

Excretion of breath and flatus gases by humans consuming high-fiber diets.

The abilities of dietary fibers to promote the excretion of intestinal fermentation gases were evaluated in five healty men. Flatus and breath gases were collected and analyzed during 3 days of each 9-day metabolic period. Responses to feeding xylan, pectin, cellulose and corn bran were compared to a fiber-free formula diet. Generally, hydrogen production increased throughout the day, whereas methane production remained more constant. Methane excretion was greater while consuming the xylan and pectin diets than while consuming the other diets. These two purified fibers also caused higher flatus volume, hydrogen and carbon dioxide excretion. Cellulose and corn bran generally resulted in breath and flatus gas excretion at levels equivalent to fiber-free diets. Considerable variation was noted in the response of subjects to an individual diet. However, in most cases 2-5 days of frequent and daily consumption of the diets was necessary to establish a relatively constant level of gas excretion. This time may represent the period of microbial and enzymatic adaptation to the new dietary constituents.

Adult↗