Fate of mucilage cell wall polysaccharides during coffee fermentation.

Effects of a 20-h fermentation on cell wall polysaccharides from the mucilage of pulped coffee beans were examined and compared to those of unfermented beans, on alcohol insoluble residues (AIRs), their hot-water-soluble crude pectic substances (PECTs), and their hot-water-insoluble residues (RESs). Yields and compositions were very similar: AIRs, which consisted of approximately 30% highly methylated pectic substances, approximately 9% cellulose, and approximately 15% neutral noncellulosic polysaccharides, exhibited no apparent degradation. However, PECTs from fermented beans were shown to have undergone a slight reduction of their intrinsic viscosity and weight-average molecular weight by capillary viscosimetry and high-performance size-exclusion chromatography. After fermentation, hot-water-insoluble pectic substances of RES exhibited partial de-esterification. Removal of coffee bean mucilage by natural fermentation seems to result from a restricted pectolysis, the mechanism of which remains to be elucidated.     

Turnover of white asparagus cell wall polysaccharides during postharvest storage.

The main polysaccharides involved in asparagus cell wall turnover have been identified. Homogalacturonans are lost from both the apical and basal sections of the spear. Galactans are mobilized from the cellulose residue of the apical section and recovered in the KOH-soluble fractions while they are lost from the cellulose residue of the basal section. Xyloglucans are incorporated in the apical region and degraded from the basal one. Cellulose is incorporated in the basal region and lost from the apical one, and acidic xylans are incorporated in high amounts in the basal section of the spear.     

Demonstration of pectic polysaccharides in cork cell wall from Quercus suber L

Scanning electron microscopy (SEM) and chemical analysis were used to observe the cell wall changes that occur in cork with "mancha amarela", when compared to a standard cork. To mimic the microbial attack exhibited in cork with mancha amarela, the standard cork was treated enzymatically with commercial pectinase and hemicellulase preparations. The tissues treated with pectinase were comparable with those attacked with mancha amarela. Both were composed by deformed and wrinkly cells and exhibited cell wall separation at the middle lamella level, which suggests solubilization/removal of the pectic polysaccharides. The cork cell wall material, prepared as alcohol-insoluble residue, was fractionated by hot water (Pect(H)()2(O)) and hot dilute acid (Pect(acid)). The relatively large amount of hexuronic acid and the occurrence of Ara in the SPect(H)()2(O) and SPect(acid) allow to confirm, as far as we know, for the first time the presence of pectic polysaccharides in the cell walls of cork from Quercus suber L. They accounted for ca. 1.5% of the cork and may consist of polymers with long side chains of arabinosyl residues. These polymers have to be taken into account in any realistic model of the cork cell wall. Cork with mancha amarela contained a smaller amount of pectic polysaccharides (ca. 0.5%), which confirms that the cellular separation observed by SEM is related to the degradation/removal of the middle lamella pectic polysaccharides.     

Perspectives of folate biofortification of cereal grains

One sixth of the world’s population is suffering from hidden hunger that indicates a gross malnutrition particularly among children and women of third world countries. The deficiency of micro nutrients, especially iron (Fe) causes a number of ailments such as megaloblastic anemia and neural tube defects in poor population. There is a dire need to supplement iron in the diet. Current efforts implicate fortification of wheat flour and other grains with different iron formulations such as ethylenediaminetetraacetic acid (EDTA), FeSO₄ and elemental iron. However, all such interventions are not sustainable due to logistic and quality assurance problems in resource-limited settings. For a long term solution, development of crop plants with increased micronutrients and iron bioavailability is essential. Therefore, biofortification of cereal grains using translational genomics approaches for enhancement of folate through genome editing in cereals is inevitable to mitigate the folate deficiency in poor remote population in a cost effective manner.     

Enzymatic degradation of cell wall polysaccharides from mango (Mangifera indica L.) puree

Ripe mango puree (Smith cultivar) was treated with fungal polysaccharidases containing pectinolytic, hemicellulolytic, and cellulolytic activities for 2 h at 50 degrees C. A loss of 30% of the cell wall material (CWM) was measured. CWM polysaccharides were hydrolyzed to varying degrees: 88, 65, and 65% of, respectively, galacturonic acid-, arabinose-, and rhamnose-containing polymers were hydrolyzed, whereas 50% of cellulose was degraded. After 30 min of treatment, the ethanol precipitation test on the serum was negative, indicating that pectic substances were rapidly hydrolyzed. Oligogalacturonic acids (degree of polymerization, 1-12) were observed in the serum. A viscosity drop of 90% was measured after 2 h, confirming the dominant role of pectic substances in puree viscosity.     

Olive fruit cell wall: degradation of pectic polysaccharides during ripening

Olive fruits at three stages of ripening (green, cherry, and black) have been studied. After cell wall isolation, the compositions of the cell wall and that of the phosphate-soluble polysaccharides were determined. In cell walls, decreases in arabinose, xylose, glucose, and uronic acid levels were observed, together with a slight increase in mannose on ripening. At the beginning of ripening, fragments of pectic polymers were the major constituents of the phosphate-soluble fraction, with the hemicellulosic ones increasing toward the end of the process. The molecular weight of the fragments solubilized was approximately 6 kDa. After cell wall fractionation, the pectic polysaccharides soluble in imidazole and sodium carbonate were also studied. In both fractions, between the green and cherry stages of ripening, a significant loss of homogalacturonans took place. Between the cherry and black stages of ripening, rhamnogalacturonan side chains were also released in addition to homogalacturonans. In any of the pectic fractions, changes in apparent molecular weight were quantified.     

Anatomy and cell wall polysaccharides of almond (Prunus dulcis D. A. Webb) seeds

The anatomy of Prunus dulcis was analyzed by applying several differential staining techniques and light microscopy. Prunus dulcis seed has a thin and structurally complex seed coat, with lignified cellulosic tissue. The embryo has two voluminous cotyledons. Cotyledon cells have a high number of protein and lipid bodies, some of which have phytin. The provascular tissue, located in the cotyledons, is oriented in small bundles perpendicular to the transverse embryonic axis. Prunus dulcis cell wall material is very rich in arabinose (45 mol %). Glucose (23%), uronic acids (12%), and xylose (12%) are also major sugar components. The polymers obtained from the imidazole and Na(2)CO(3) extracts contain mainly pectic substances rich in arabinose, but the sugar content of these extracts was very low. The majority of the pectic substances (also rich in arabinose) was recovered with the KOH extracts. These extracts, with high sugar content, yielded also xyloglucans and acidic xylans. The 4 M KOH + H(3)BO(3) extracts yielded polysaccharides rich in uronic acids and xylose and very rich in arabinose, accounting for 27% of the cell wall material.     

Combined enzymatic and high-pressure processing affect cell wall polysaccharides in berries

The effect of high-pressure processing (HPP) on cell wall polysaccharides in berries was investigated. HPP decreased the degree of methyl esterification (DM), probably by activation of pectin methyl esterase (PME), and improved the extractability of pectins. When commercial enzyme mixtures were added to mashed berries, a synergistic effect was observed between treatment with commercial enzymes and HPP. Compared to treatment at atmospheric pressure, pectic polysaccharides were degraded to a larger extent when HPP was used. In contrast, hemicelluloses were hardly affected by the added enzymes when HPP was included, although they were degraded during similar treatment at atmospheric pressure. Additionally, the activity of rhamnose-releasing enzymes present in minor quantities might be enhanced after HPP, resulting in a decrease of rhamnose in the polymeric cell wall material. These results exploring the effect of HPP at representative conditions clearly point out the potential of HPP for polysaccharide modification.     

Olive fruit cell wall: degradation of cellulosic and hemicellulosic polysaccharides during ripening

Cellulose and hemicelluloses obtained from the cell walls of partially depectinated olives have been studied at three stages of ripening (green, cherry, and black). Hemicelluloses were fractionated into two groups, the amounts of which diminished during ripening: those soluble in 4% KOH diminished between the cherry and black stages, whereas those soluble in 24% KOH did so between the green and cherry stages. Arabinoxylans, xyloglucans, and homo- and/or rhamnogalacturonans to a lesser extent were present in these fractions. After ion exchange and size exclusion chromatographies, decreases in the molecular weights of hemicelluloses, mainly in the neutral fractions, were observed. The amount of cellulose also decreased, but at the second stage of the ripening process. Approximately 2 mg/fruit of glucose was lost from cellulose, and the amount of uronic acids increased (0.23 mg/fruit).     

Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria

New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied. Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple. All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested. Bacteroides spp. are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides. Bifidobacterium spp. are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides. Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro. These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains.     

Cell wall polysaccharides from chalkumra (Benincasa hispida) fruit. Part I. Isolation and characterization of pectins

Pectic polysaccharides were obtained from chalkumra (Benincasa hispida) fruit by sequential extraction with ammonium oxalate (fraction BOX), dilute acid (fraction BHCl), and cold dilute alkali (fraction BOH). The highest yield of polysaccharides was obtained with oxalate and HCl. BOX was enriched in partly methyl-esterified galacturonic acid, whereas BHCl and BOH contained mostly galactose. All of the extracts showed similar elution patterns in size exclusion chromatography although the intrinsic viscosities (eta) were different (132 +/- 6, 100 +/- 5, and 285 + 10 mL/g for BOX, BHCl, and BOH, respectively). From fractionation by anion exchange chromatography, homogalacturonan (as seen from sugar analysis and Fourier transform infrared spectrum) accounted for more than half of BOX and 11% of BHCl. Methylation analyses and hydrolysis of BHCl with endo-beta-(1-->4)-d-galactanase showed the presence of beta-(1-->4)-d-galactan. The neutral galactan represented more than 76% of BHCl and approximately 40% of BOH. The other polysaccharides were complex galactans in BOH and an acidic arabinan (<1%) in BOX and BHCl.     

Liquid chromatographic determination and stability of the Fusarium mycotoxin moniliformin in cereal grains

Moniliformin is a mycotoxin produced by Fusarium subglutinans and other Fusarium species. A rapid, liquid chromatographic method for its determination in corn and wheat is described. Samples are extracted in acetonitrile-water (95 + 5); following defatting with n-hexane, an aliquot of the extract is evaporated and cleaned up on small C18 and neutral alumina columns successively. Reverse-phase liquid chromatography (LC) is conducted on a C18 column with 10 or 15% methanol or acetonitrile in aqueous ion-pair reagent as mobile phase, with detection by ultraviolet absorption at 229 and 254 nm. Average recoveries of moniliformin (potassium salt) added to ground corn and wheat at levels of 0.05-1.0 micrograms/g were 80% (n = 20) and 85% (n = 12), respectively, and the limit of detection was ca 0.01-0.18 micrograms/g, depending on LC conditions. Analysis of 24 samples of wheat, 4 samples of rye, and 12 samples of corn showed moniliformin in only 2 corn samples (0.06 and 0.2 micrograms/g). Moniliformin was also detected in a sample of artificially damaged (slashed) corn (0.2 micrograms/g) and selected kernels of corn that were field-inoculated with F. subglutinans and F. moniliforme (50 micrograms/g and 0.5 micrograms/g, respectively). In stability studies, moniliformin (potassium salt, 1 microgram/g) in ground corn and ground wheat heated at 50, 100, and 150 degrees C for 0.5-2 h decomposed moderately, e.g., 55% remained in corn after 0.5 h at 100 degrees C.     

Deposition of cell wall polysaccharides in wheat endosperm during grain development: Fourier transform-infrared microspectroscopy study

The time course and pattern deposition of the cell wall polysaccharides in the starchy endosperm of wheat (Triticum aestivum cv. Recital) during grain development was studied using Fourier transform infrared microspectroscopy (micro-FTIR). Three stages of grain development identified as key stages for cell wall construction were retained as follows: the end cellularization, the beginning of cell differentiation, and the beginning of maturation. Micro-FTIR revealed that beta-(1-->3),(1-->4) glucans and arabinoglactan proteins are the main cell wall components of endosperm at the end of the cellularization stage, whereas arabinoxylans (AX) appeared only at the cell differentiation stage. Past the differentiation stage, FTIR spectra were dominated by AX features. Cell walls at the beginning of cell differentiation and at endosperm maturation could be distinguished by spectral features that were ascribed to AX substitution. AX appeared more substituted at the beginning of cell differentiation. Moreover, a difference in the degree of AX substitution was found between peripheral and central parts of the grain at the cell differentiation stage; AX in central cells was less substituted. Thus, dramatic changes in endosperm cell wall composition were detected during wheat grain development with respect to both the relative occurrence of individual constituents and the fine structure of the AX.     

Determination of ochratoxin A in animal feed and cereal grains by liquid chromatography with fluorescence detection

A liquid chromatographic (LC) method is described for determination of ochratoxin A in animal feeds and cereal grains. Samples are initially extracted with chloroform-ethanol (8 + 2) and 5% acetic acid in water. Extracts are purified using a silica gel cartridge followed by a cyano cartridge. The samples are evaporated, diluted to a known volume, and analyzed using a 10 cm column of 3 micron C18 and a fluorescence detector. The method was applied to a variety of animal feeds and cereal grains at levels of 1.0-0.005 ppm added ochratoxin A. The overall recovery was 90.6% +/- 3.6.