Activity and process stability of purified green pepper (Capsicum annuum) pectin methylesterase

Pectin methylesterase (PME) from green bell peppers (Capsicum annuum) was extracted and purified by affinity chromatography on a CNBr-Sepharose-PMEI column. A single protein peak with pectin methylesterase activity was observed. For the pepper PME, a biochemical characterization in terms of molar mass (MM), isoelectric points (pI), and kinetic parameters for activity and thermostability was performed. The optimum pH for PME activity at 22 degrees C was 7.5, and its optimum temperature at neutral pH was between 52.5 and 55.0 degrees C. The purified pepper PME required the presence of 0.13 M NaCl for optimum activity. Isothermal inactivation of purified pepper PME in 20 mM Tris buffer (pH 7.5) could be described by a fractional conversion model for lower temperatures (55-57 degrees C) and a biphasic model for higher temperatures (58-70 degrees C). The enzyme showed a stable behavior toward high-pressure/temperature treatments.     

Effect of germination on the carbohydrate composition of the dietary fiber of peas (Pisum sativum L.)

The effect of different conditions of pea germination on dietary fiber (DF) composition was studied. Insoluble dietary fiber (IDF) and soluble dietary fiber (SDF) were subjected to acid hydrolysis, and the resultant neutral sugars, uronic acids, and Klason lignin were quantified. Germinated peas exhibited significantly higher contents of total dietary fiber (TDF) than the raw sample, due to the increases of both DF fractions. Under darkness conditions, germination exhibited the highest contents of IDF and SDF. Decreasing IDF/SDF ratios showed that the carbohydrate changes did not take place to the same extent during germination, the SDF fraction being the most affected. The detailed chemical composition of fiber fractions reveals increases of cellulose in the IDF of germinated samples, whereas SDF exhibits a decrease of pectic polysaccharides and also increases of polysaccharides rich in glucose and mannose. The DF results were corroborated by a comparative examination of the cell wall carbohydrate composition.     

Influence of disaccharide structure on prebiotic selectivity in vitro

To obtain structure-function information of a range of carbohydrates, which are available only in very small quantities, an in vitro fermentation method using 7 mg of carbohydrate, 0.7 mL of basal medium, and 1% (w/v) of fecal bacteria was validated against a pH-controlled batch culture with 150 mL of basal medium and 1.5 g of test carbohydrate. This method was used to determine the influence of different glycosidic linkages and monosaccharide compositions of disaccharides on the selectivity of microbial fermentation. A prebiotic index (PI) was calculated for each disaccharide. Generally, disaccharides with linkages of 1-2, 1-4, and 1-6 generated a high PI score, with kojibiose and sophorose showing the greatest values (21.62 and 18.63, respectively). Apart from 6alpha-mannobiose, mannose-containing disaccharides gave a low PI due to low numbers of bifidobacteria and lactobacilli and an increase in bacteroides. The structure-function information obtained in this study may lead to a predictive understanding of how specific structures are fermented by the human gut microflora.     

Nutritional evaluation of pea (Pisum sativum L.) protein diets after mild hydrothermal treatment and with and without added phytase

The effect of mild hydrothermal treatment and the addition of phytase under optimal conditions (pH 5.5, 37 degrees C) on the nutritive utilization of the protein of pea (Pisum sativum L.) flour was studied in growing rats by examining the chemical and biological balance. Mild hydrothermal treatment produced reductions of 83, 78, and 72%, respectively, in the levels of alpha-galactosides, phytic acid, and trypsin inhibitors and also produced a significant increase in the digestive utilization of protein. The additional fall in the levels of phytic acid caused by the addition of phytase did not lead to a subsequent improvement in the digestive utilization of protein. The mild hydrothermal treatment of pea flour produced a significant increase in the metabolic utilization of protein and carbohydrates, which was reflected in the protein efficiency ratio and food transformation growth indices. These effects were not observed in the phytase-supplemented pea diet.     

2-Furoylmethyl amino acids and hydroxymethylfurfural as indicators of honey quality

Determination of changes in 2-furoylmethyl amino acids and hydroxymethylfurfural during the storage of four honey samples at 25 and 35 degrees C during 12 months was achieved to assess the potential use of both parameters, singly or in combination, as quality indicators. 2-Furoylmethyl amino acids increased during storage at both temperatures, whereas hydroxymethylfurfural only presented slight variations during storage at 25 degrees C but increased noticeably at 35 degrees C. The study of 2-furoylmethyl amino acids in 49 commercial honeys revealed that 2-furoylmethyl lysine (furosine) was present in all samples, whereas 2-furoylmethyl derivatives of arginine, GABA, and proline were only present in seven samples. Hydroxymethylfurfural can be considered as a good indicator of heat treatments applied to honey samples, whereas 2-furoylmethyl amino acids can be used as suitable markers of the storage period. The use of both parameters can be useful to detect adulteration with invert syrups, excessive heat treatments, or prolonged storage of honey samples.     

Organophosphorus pesticide residues in Mexican commercial pasteurized milk

A study was conducted to measure residues of 13 organophosphorus (OP) pesticides, widely used as dairy cattle ectoparasiticides or in crops used for animal feed, in homogenized and pasteurized Mexican milk samples. Four different milk brands with high distribution were collected biweekly during a 12 month period (n = 96) in supermarkets. OP pesticide residues were measured by gas chromatography with a flame photometric detector. Approximately 39.6% of the samples contained detectable levels of OP pesticide residues. Eight samples contained residues exceeding established maximum residue limits (MRL), and the OP pesticides present in these samples were dichlorvos (five samples), phorate, chlorpyrifos, and chlorfenvinphos (one sample, respectively). Average residues of 13 OP pesticides measured were below established MRLs ranging between 0.0051 and 0.0203 ppm.     

Ozone Biomonitoring with Bel-W3 Tobacco Plants in the City of Valencia (Spain)

A biomonitoring study using the ozone-sensitive bioindicator plant Nicotiana tabacum cv. Bel-W3 was conducted in the city of Valencia (eastern Spain) and surrounding areas in 2002. Plants were exposed to ambient air at seven sites, including four traffic-exposed urban sites, a large urban garden and a suburban and a rural station, for six consecutive 2-week periods using highly standardised methods. Foliar injury was registered at all stations in at least one of the exposure periods. The urban stations submitted to intense traffic showed lower ozone injury than the less traffic-exposed stations. Strong changes in the intensity of ozone injury were observed for the different exposure periods. Leaf injury was significantly related to both mean ozone values (24 and 12 h means) and cumulative exposure indices (AOT20, AOT40). However, correlation strength was moderate (r _s?=?0.39 to 0.58), suggesting that the plant response to ozone was modified by environmental factors. The use of sensitive bioindicators like tobacco Bel-W3 in cities provides complementary information to that of continuously operating air quality monitors, as the impact of ambient ozone levels is directly measured.     

Genetic diversity of some accessions of Cucurbita maxima from Spain using RAPD and SBAP markers

Increasing the knowledge of the molecular diversity of a crop is essential for extending its genetic base, identifying cultivars and selecting parental varieties for breeding programs. In this sense, Cucurbita maxima Duch. is poorly characterised. Nineteen accessions of this species and 8 related Cucurbita accessions were included in a genetic diversity analysis. For this purpose, Random Amplified Polymorphic DNA markers (RAPDs), which analyse neutral variability, and Sequence-Based Amplified Polymorphism (SBAPs), which preferentially amplify coding regions of the genome, were used. While the UPGMA cluster and the principal coordinates analysis obtained using RAPDs did not group the different accessions according either to fruit morphological criteria or to passport data (origin and agro-climatic conditions), the principal coordinates analysis obtained using SBAPs grouped the different pumpkin accessions fundamentally according to the type of use (human consumption, animal fodder or ornamental). This passport trait is reported to be associated with agronomic breeding characters of interest. The usefulness of both types of markers for discriminating accessions of breeding interest is discussed.     

Improvement of table grapes quality and safety by the combination of modified atmosphere packaging (MAP) and eugenol, menthol, or thymol

Table grape is a nonclimacteric fruit that shows a rapid loss of quality during storage and is very susceptible to colonization by fungi, especially Botrytis cinerea, which is considered the most important disease of this commodity. To solve this problem, synthetic fungicides have been used, although legal restrictions and consumer's concern demand the search of other safe means. In the present paper, and as an alternative of synthetic fungicides, an active packaging to improve MAP effectiveness on preserving table grape (cv. Crimson Seedless) quality and safety was developed by the addition of 0.5 mL of eugenol, thymol, or menthol inside the packages. Packages were stored at 1 degree C for 35 days. The final gas composition inside the packages was 1.4-2.0 and 10.0-14.5 kPa of CO(2) and O(2), respectively, with no significant slight differences among treatments. Results showed that the addition of eugenol, thymol, or menthol improved the beneficial effect of MAP in terms of delaying weight loss and color changes, retarding degrees Brix/acidity ratio evolution, and maintaining of firmness. Thereafter, these treatments showed additional benefit in terms of delayed rates of rachis deterioration and berry decay. Finally, the total viable counts for both mesophilic aerobics and especially yeast and molds were significantly reduced in the grapes packaged with the natural antimicrobial compounds. All of the above effects led to maintenance of table grape quality and safety for longer storage periods (3 additional weeks as compared to controls under MAP only).     

Changes in the chemical composition of water percolating through the soil profile in a moderately polluted catchment

Throughfall (TF), stemflow (SF), soil solution below the organic layer (SS_org) and at 50 cm depth (SS₅₀), and output with stream water (SW) were measured and analyzed for four years in a moderately polluted forest catchment in southern Poland. The input of water with stemflow was ca. 6% of input with TF. However, due to higher concentrations of most ions in SF, the input of most elements with SF was from 8% to 9%. Sulphate (SO₄ ^2–), chloride (Cl^–) and magnesium (Mg²⁺) were the only ions steadily increasing in concentrations in water percolating through the soil profile. Nitrogen reached the forest floor mainly as ammonium (NH₄ ⁺). In the soil organic layer the NH₄ ⁺ concentration decreased, while concentrations of nitrate (NO₃ ^–) and hydrogen (H⁺) increased, probably due to nitrification. For NO₃ ^–, sodium (Na⁺) and calcium (Ca²⁺), the highest concentrations were found in SS_org and SW. This indicates both efficient cycling in the biotic pool of the ecosystem and intensive weathering processes in the mineral soil below the plant rooting zone. The latter was especially pronounced for Mg and Ca. Concentrations of zinc (Zn), lead (Pb) and cadmium (Cd) were the highest in SS_org and SS₅₀. As this was accompanied by a low pH and constant input of H⁺, NH₄ ⁺ and heavy metal ions to the catchment area, it may pose a serious threat to forest health.