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At least 19 recordsLinked to original sources

[Thermodynamic approach to the selection of polyuronide sequestrants for the protection of the human body from toxic metal ions. Interactions of polyuronides with strontium and calcium ions].

Selectivity of polyuronide sequestrants (pectate, alginates of various uronide composition) in respect to Sr2+ and Ca2+ ions has been evaluated in terms of thermodynamic affinity. It is suggested that there is no point in the use of pectate as a Sr(2+)-binding agent because at initial stages of reaction it reveals higher affinity to Ca2+ ions in comparison to Sr2+ ions. Contrary to pectate, alginates under similar conditions have higher affinity to Sr2+ ions. It is shown that these ions are bound only by blocks of L-guluronic acid residues in alginate macromolecules. The results obtained lend support to the advisability of the use of alginate preparations with the high content of L-guluronic acid residues for the excretion of Sr2+ ions from human body.

Alginates↗

[Thermodynamic approach to the selection of polyuronide sequestrants for protection of the human body from toxic metal ions. Interactions of polyuronides with lead ions].

Binding isotherms of Pb2+ ions with potassium pectate and potassium alginate with relatively low content of blocks of L-guluronic residues (20%) have been determined. Interactions of Pb2+ ions with polyuronides studied is cooperative. Maximum values of binding constants are an order of magnitude higher than previously determined ones for Ca2+ and Sr2+ ions. Along with ion-coordination ("stoichiometric") interactions, alginate is typified by so-called extra-stoichiometric binding of Pb2+ ions, which presumably proceeds by a coprecipitation mechanism. Limitations of the thermodynamic approach to the selection of sequestrants for human body protection from toxic metal ions are discussed.

Alginates↗

Polyuronides in Avocado (Persea americana) and Tomato (Lycopersicon esculentum) Fruits Exhibit Markedly Different Patterns of Molecular Weight Downshifts during Ripening.

Avocado (Persea americana) fruit experience a rapid and extensive loss of firmness during ripening. In this study, we examined whether the chelator solubility and molecular weight of avocado polyuronides paralleled the accumulation of polygalacturonase (PG) activity and loss in fruit firmness. Polyuronides were derived from ethanolic precipitates of avocado mesocarp prepared using a procedure to rapidly inactivate endogenous enzymes. During ripening, chelator (cyclohexane-trans-1,2-diamine tetraacetic acid [CDTA])-soluble polyuronides increased from approximately 30 to 40 [mu]g of galacturonic acid equivalents (mg alcohol-insoluble solids)-1 in preripe fruit to 150 to 170 [mu]g mg-1 in postclimacteric fruit. In preripe fruit, chelator-extractable polyuronides were of high molecular weight and were partially excluded from Sepharose CL- 2B-300 gel filtration media. Avocado polyuronides exhibited marked downshifts in molecular weight during ripening. At the postclimacteric stage, nearly all chelator-extractable polyuronides, which constituted from 75 to 90% of total cell wall uronic acid content, eluted near the total volume of the filtration media. Rechromatography of low molecular weight polyuronides on Bio-Gel P-4 disclosed that oligomeric uronic acids are produced in vivo during avocado ripening. The gel filtration behavior and pattern of depolymerization of avocado polyuronides were not influenced by the polyuronide extraction protocol (imidazole versus CDTA) or by chromatographic conditions designed to minimize interpolymeric aggregation. Polyuronides from ripening tomato (Lycopersicon esculentum) fruit extracted and chromatographed under conditions identical with those used for avocado polyuronides exhibited markedly less rapid and less extensive downshifts in molecular weight during the transition from mature-green to fully ripe. Even during a 9-d period beyond the fully ripe stage, tomato fruit polyuronides exhibited limited additional depolymerization and did not include oligomeric species. A comparison of the data for the avocado and tomato fruit indicates that downshifts in polyuronide molecular weight are a prominent feature of avocado ripening and may also explain why molecular down-regulation of PG (EC 3.2.1.15) in tomato fruit has resulted in minimal effects on fruit performance until the terminal stages of ripening.

Journal Article↗

Effect of Antisense Suppression of Endopolygalacturonase Activity on Polyuronide Molecular Weight in Ripening Tomato Fruit and in Fruit Homogenates.

Fruit of tomato (Lycopersicon esculentum Mill.) in which endopolygalacturonase (PG) activity had been suppressed to <1% of wild-type levels were slightly firmer than nontransgenic controls later in ripening. Enzymically inactive cell walls were prepared from these ripening fruit using Tris-buffered phenol. When extracted with chelator followed by Na2CO3, the amounts of pectin solubilized from cell walls of nontransgenic control or from transgenic antisense PG fruit were similar. Size-exclusion chromatography analysis showed that, relative to controls, in antisense PG fruit polyuronide depolymerization was delayed in the chelator-soluble fraction throughout ripening and reduced in the Na2CO3-soluble fraction at the overripe stage. Reduced pectin depolymerization rather than altered extractability thus may have contributed to enhanced fruit firmness. Substantially larger effects of suppressed PG activity were detected in tomato fruit homogenates processed to paste. In control paste the majority of the polyuronide was readily soluble in water and was very highly depolymerized. In antisense PG paste the proportion of polyuronide solubilized by water was reduced, and polyuronides retained a high degree of polymerization. The suppression of fruit PG activity thus has a small effect on polyuronide depolymerization in the fruit but a much larger effect in paste derived from these fruit. This indicates that in the cell wall PG-mediated degradation of polyuronide is normally restricted but that in tissue homogenates or in isolated cell walls this restriction is removed and extensive pectin disassembly results unless PG is inactivated.

Journal Article↗

Analysis of the molecular size of tomato (Lycopersicon esculentum Mill) fruit polyuronides by gel filtration and low-speed sedimentation equilibrium.

The cell-wall structures of tomato (Lycopersicon esculentum Mill) and other fruit are intimately linked with the nature of their polyuronides. Cell-wall polyuronides from unripe and ripe tomato fruit were isolated and purified and their molecular size and molecular-size distributions were compared. It was demonstrated that there is a considerable decrease in the weight-average Mr upon ripening (from 160,000 +/- 10,000 to 96,000 +/- 4000) and a corresponding increase in polydispersity, particularly at the low-Mr end of the distribution. The estimates of polyuronide molecular size and molecular-size distribution were obtained without the need for polyuronide standards of known Mr by using gel-filtration chromatography combined with the absolute method of low-speed sedimentation equilibrium.

Centrifugation, Density Gradient↗

Application of polyuronides for removing heavy metals from vegetable oils. III. Application of alginic acid, pectic and pectinic acids for demetalization of hydrogenated sunflower oil.

Laboratory experiments have been carried out for the removal of heavy metals from hydrogenated vegetable oils using hydrated polyuronides (degree of swelling from 4 to 12.8 ml/g) such as alginic acid, pectic and pectinic acids. The effect of the type of polyuronide, degree of esterification and oil treatment on the degree of demetalization has been studied. It has been shown that with increase in the degree of esterification of the polyuronide the efficiency of demetalization decreases. The second and third treatment of the hydrogenated oil with pectinic acid resulted in a high degree of heavy metal removal. The possibility of efficient demetalization of hydrogenated oils by treatment with water solutions of pectinic acids has also been demonstrated. The degree of metal ion removal increases with decreasing concentration of pectinic acids in the water solution.

Adsorption↗

A bound form of silicon in glycosaminoglycans and polyuronides.

Silicon was found to be a constituent of certain glycosaminoglycans and polyuronides, where it occurs firmly bound to the polysaccharide matrix. 330-554 ppm of bound Si were detected in purified hyaluronic acid from umbilical cord, chondroitin 4-sulfate, dermatan sulfate, and heparan sulfate. These amounts correspond to 1 atom of Si per 50,000-85,000 molecular weight or 130-280 repeating units. 57-191 ppm occur in chondroitin 6-sulfate, heparin, and keratan sulfate-2 from cartilage, while hyaluronic acids from vitreous humor and keratan sulfate-1 from cornea were Si-free. Large amounts of bound Si are also present in pectin (2580 ppm) and alginic acid (451 ppm). The bound Si is not dialyzable, does not react with ammonium molybdate, is not liberated by autoclaving or 8 M urea, and is stable against weak alkali and acid. Strong alkali and acid hydrolyze the Si-polysaccharide bond. Free, direct-reacting, dialyzable silicate is obtained. Enzymatic hydrolysis of hyaluronic acid or pectin does not liberate silicic acid, but leads to products of low molecular weight still containing Si in bound form. It is concluded that Si is present as a silanolate, i.e., an ether (or esterlike) derivative of silicic acid, and that R(1)-O-Si-O-R(2) or R(1)-O-Si-O-Si-O-R(2) bridges play a role in the structural organization of glycosaminoglycans and polyuronides. Thus, Si may function as a biological crosslinking agent and contribute to architecture and resilience of connective tissue.

Animals↗

Polyuronide biosynthesis by cell-free extracts of Mucor rouxii.

Cell-free extracts from Mucor rouxii contain enzymes that catalyse the synthesis of uridine diphosphate glucuronic acid (UDPGlcA) from UDPglucose and the incorporation of glucuronic acid from UDPGlcA into polymer(s). Two different polyuronide fraction isolated from the cell walls of this fungus were used as primers. Mucoran, a heteropolymer, was much more efficient than mucoric acid, which is largely a homopolymer of D-glucuronic acid. The primer ability of native cell walls was comparable to that of mucoric acid. Most of the glucuronosyltransferase activity in the cell-free extract was found in a 20000 g particulate fraction. Optimum pH for polyuronide synthesis was 7.0. Mn2+ or Mg2+ stimulated incorporation of GlcA. The products synthesized from mucoric acid and mucoran primers were different and yielded different disaccharides upon hydrolysis.

Biopolymers↗

Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening.

Tomato fruit ripening is accompanied by extensive degradation of pectic cell wall components. This is thought to be due to the action of a single enzyme, polygalacturonase, whose activity is controlled, at least in part, at the level of gene expression. At the onset of tomato fruit ripening, polygalacturonase enzyme activity, mRNA levels, and relative rate of gene transcription all increase dramatically. To elucidate the role of polygalacturonase during tomato fruit ripening, we utilized a pleiotropic genetic mutation, rin, that blocks many aspects of ripening, including the activation of polygalacturonase gene transcription. The polygalacturonase structural gene was ligated to a promoter that is inducible in mature rin fruit and inserted into the fruit genome, and plants were regenerated. This allowed expression of the polygalacturonase gene in transgenic rin fruit at a time corresponding to ripening in wild-type fruit. Expression of this gene resulted in the accumulation of active polygalacturonase enzyme and the degradation of cell wall polyuronides in transgenic rin fruit. However, no significant effect on fruit softening, ethylene evolution, or color development was detected. These results indicate that polygalacturonase is the primary determinant of cell wall polyuronide degradation, but suggest that this degradation is not sufficient for the induction of softening, elevated rates of ethylene biosynthesis, or lycopene accumulation in rin fruit.

Base Sequence↗

Thermodynamic approach to the selection of polyuronide sequestrants for preventive and medicinal nutrition.

An approach to the analysis of isotherms of cooperative binding is developed that allows to calculate approximately affinity profiles, i.e. dependencies of binding constants on binding densities. The comparison of the affinity profiles of the interactions of Ca2+ and Sr2+ ions with sodium pectate as well as sodium alginates with the different content of the blocks of alpha-L-guluronic acid residues (GG-blocks) and blocks of mixed composition showed that (1) pectate has higher affinity to both ions as compared with alginates; (2) the affinity of pectate to Ca2+ ions is comparable to its affinity to Sr2+ ions but in the case of Ca2+ ions the maximum of the affinity profile falls on the substantially lesser value of the binding density in comparison to Sr2+ ions that seems to be unfavorable from the standpoint of the use of this polyuronide in preventive or medicinal nutrition; (3) the affinity of both alginates to Sr2+ ions at relatively low binding densities exceeds their affinity to Ca2+ ions; (4) the affinity of alginates to Sr2+ ions increases with the increase in the content of GG-blocks, whereas the affinity to Ca2+ ions practically does not depend on the composition of the alginate; (5) the maximum binding density of Sr2+ ions to the alginates approximately corresponds to the content of GG-blocks. These results corroborate the practice of the use of alginates rich in residues of alpha-L-guluronic acid for the removal of radioactive strontium from the digestive tract. The binding constants of Pb2+ ions to pectate over a wide range of binding densities are more than an order of magnitude greater than those of alkali-earth metal ions.

Alginates↗

Two distinct classes of polyuronide from the cell walls of a dimorphic fungus, Mucor rouxii.

Polyuronides were extracted from purified yeast and mycelial walls of Mucor rouxii by sequential treatments with lithium chloride and potassium hydroxide and were fractionated by ion-exchange chromatography on DEAE-Sephadex. Two polymers (I and II) of different acidity were found in both wall types. Polymer I contained D-glucuronic acid, L-fucose, D-mannose, and much smaller amounts of D-galactose. Yeast and mycelial polymer I had similar uronic acid contents but differed in their neutral sugar compositions and molecular weights. Polymer II from both cell types contained largely D-glucuronic acid and had similar molecular weights. On partial acid hydrolysis, both polymers I and II gave rise to insoluble glucuronans which appeared to be homopolymeric. One-third of the total uronosyl residues of polymer I, and almost all of the uronosyl residues of polymer II, were present in homopolymeric segments. However, homopolymers derived from polymers I and II may not be identical.

Cell Wall↗

The polyuronides.

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Uronic Acids↗