Oligosaccharides Raffinose Research Articles

Influence of Extraction Solvent and Temperature on the Quantitative Determination of Oligosaccharides from Plant Materials by High-Performance Liquid Chromatography

The effect of extraction solvents and temperatures on extraction yields of monosaccharides, sucrose, and raffinose oligosaccharides from plant materials was investigated. Toasted soybean meal, cott...

Differential Accumulation of Carbohydrates in Alfalfa Cultivars of Contrasting Winterhardiness

Soluble sugar composition and starch reserves are significantly altered during cold hardening of alfalfa (Medicago sativa L.). We characterized the relationship between carbohydrate composition of the crown and freezing tolerance in cultivars of contrasting winterhardiness during their acclimation to low temperature under controlled (two cultivars) and natural hardening conditions (three cultivars in 1991–1992 and six cultivars in 1992–1993). During hardening under environmentally controlled conditions, freezing tolerance and crown levels of soluble sugars increased significantly with a concomitant decrease in starch. Differences in freezing tolerance between a cold‐tolerant and a cold‐sensitive cultivar were closely associated to the accumulation of the oligosaccharides raffinose and stachyose but were not related to the levels of sucrose. Incubation of prehardened plants at subzero temperatures (−2°C) increased freezing tolerance and promoted higher accumulations of sucrose, raffinose, and stachyose and further reduction in starch levels. Under natural hardening conditions, sucrose concentration in crowns was poorly related to the differences in midwinter levels of freezing tolerance between cultivars. Stachyose and raffinose accumulated later in fall than sucrose and reached higher levels in winterhardy than in nonhardy cultivars. Our results show that the accumulation of sucrose, stachyose, and raffinose and the decrease in glucose, fructose, and starch levels are temporally related to the development of freezing tolerance in alfalfa. However, differences in the maximum level of freezing tolerance between nonhardy and winterhardy cultivars are better related to the capacity of the plants to accumulate stachyose and raffinose than to accumulate sucrose.

Studies on the Carbohydrate Content of Breadfruit (Artocarpus communis Forst) From South‐Western Nigeria

AbstractThe proximate composition of the peel, pulp and core of breadfruit revealed that the highest moisture, ash, protein, fat and crude fibre contents can be found in the core while the pulp contained the highest levels of carbohydrate, starch, nitrogen free extract and organic matter. Total free and reducing sugars were highest in the core and lowest in the peel. Sucrose, glucose and fructose followed a similar pattern as the reducing sugars while the flatus‐producing oligosaccharides raffinose (0.1%) and stachyose (0.05%) were present in the core. Only raffinose was present (0.05%) in both the peel and the pulp. Extracted starch from the breadfruit pulp was 58% of the total starch content on dry weight basis with minimal levels of ash, fat, protein and 98.6% starch. The extracted starch was 98% pure and contained only 2.3% damaged starch. The starch swelling and solubility properties showed a two‐stage pattern while the Brabender amylograms showed patterns very typical of starches from most normal non‐waxy cereals. The kinetics of carbohydrate breakdown under different storage conditions showed a biphasic first order, slow in the first 8h and thereafter accelerated. Carbohydrate breakdown as an indicator of fruit deterioration was highest when breadfruit was stored at room temperature, while there was only a little difference in those stored in water, formaldehyde solution or in the refrigerator.

Oligosaccharides, Antinutritional Factors, and Protein Digestibility of Dry Beans as Affected by Processing

ABSTRACTDry beans (Phaseolus vulgaris) were subjected to soaking, cooking or a combination of both prior to fermentation, and then assessed for oligosaccharides, antinutritional factors and in‐vitro protein digestibility. Results showed an important decrease in raffinose oligosaccharides and antinutritional factors. However, an increase of trypsin inhibitor and tannin contents occurred respectively in cooked or soaked‐cooked fermented beans and in raw or soaked fermented beans. Appreciable improvement in in‐vitro protein digestibility was only observed in cooked or soaked‐cooked beans. After fermentation, the largest decreases were observed in soaked‐cooked beans (92.75%) for raffinose, in cooked beans (31.57%) for phytic acid, in soaked beans (90.86%) for stachyose, and in raw beans for trypsin inhibitor (38.77%). The highest increase due to fermentation was observed in raw beans for in‐vitro protein digestibility (1.73%).

The intermediary cell: Minor-vein anatomy and raffinose oligosaccharide synthesis in the Scrophulariaceae

Minor-vein anatomy, sugar content, sugar synthesis, and translocation were studied in mature leaves of nine members of the Scrophulariaceae to determine if there is a correlation between companion-cell type and class of sugar translocated. Three types of companion cell were found: intermediary cells with extensive plasmodesmatal connections to the bundle sheath; transfer cells with wall ingrowths and few plasmodesmata; and ‘ordinary’ companion cells with few plasmodesmata and no wall ingrowths. Alonsoa warscewiczii Regal., Verbascum chaixi Vill., and Mimulus cardinalis Dougl. ex. Benth. have intermediary cells and ordinary companion cells in the minor veins. These plants synthesize large amounts of raffinose and stachyose as well as sucrose. Nemesia strumosa Benth., and Rhodochiton atrosanguineum Zucc. have both intermediary cells and transfer cells and make proportionately less raffinose oligosaccharide than the species above. In N. strumosa, a single sieve element may abut both an intermediary cell and a transfer cell. The minor veins of Asarina scandens (Cav.) Penn. have transfer cells and what appear to be modified intermediary cells that have fewer plasmodesmata than other species, and occasional wall ingrowths. Asarina scandens synthesizes little raffinose or stachyose. Cymbalaria muralis P. Gaertn et al. and Linaria maroccana Hook.f. have only transfer cells and Digitalis grandiflora Mill. has only ordinary companion cells; these species make a trace of galactinol and raffinose, but no stachyose. Translocation experiments indicate that there is long-distance movement of raffinose oligosaccharide in these plants, even when it is synthesized in very small quantities in the leaves. We conclude that intermediary cells are as distinct a cell type as the transfer cell. In contrast to transfer cells, which are specialized for uptake of solute from the apoplast, intermediary cells are specialized for symplastic transfer of photoassimilate from the mesophyll and for synthesis of raffinose oligosaccharide. This supports our contention that raffinose oligosaccharide synthesis and symplastic phloem loading are mechanistically linked (Turgeon and Gowan 1990, Plant Physiol. 94, 1244–1249). Minor-vein anatomy and sugar synthesis may be useful characters in determining the phylogenetic relationships of plants in this family.

Changes in the carbohydrate composition of legumes after soaking and cooking

Using a high-performance liquid chromatography (HPLC) method, researchers analyzed the sugar composition of chick-peas, kidney beans, and lentils at various points in the preparation and cooking process: after soaking, after “normal” cooking (ie, boiling), after pressure-cooking, and after cooked legumes had been held at 35°C for 5 hours. There was a considerable decrease in the amount of monosaccharides, disaccharides, and raffinose oligosaccharides in chick-peas and kidney beans after soaking and cooking. This change in carbohydrate composition was less pronounced when the cooking water was not drained before analysis, which was the method used when analyzing the lentils. Method of cooking (either boiling or pressure-cooking) did not have different effects on the sugar composition of chick-peas and lentils, but loss of oligosaccharides was slightly higher when kidney beans were boiled than when they were pressure-cooked. Loss of α-galactosides occurred in chick-peas and kidney beans that had been boiled and then held at 35°C for 5 hours. The HPLC analysis showed that manninotriose was not one of the oligosaccharides present in these legumes.

Stachyose and mannitol transport in olive (Olea europaea L.)

Source leaves of Olea europea L. (olive) were photosynthetically labelled with 14CO2 and then transferred to 20 mM EDTA solutions to allow exudation of phloem sap. Label in phloem sap was recovered predominantly as stachyose (50–60% of the total label) while only small amounts of label were recovered in the sugar alcohol mannitol (less than 5%). In contrast, in leaf tissues stachyose accounted for only 5% of the total leaf label while mannitol accounted for 30%. Vacuum-infiltration of exuding leaves with 1 mM p-chloromercuriphenylsulfonic acid had no effect on subsequent exudation or on the distribution of label in stachyose or mannitol in either leaf tissues or phloem exudates. In contrast, a similar treatment inhibited the uptake of exogenously supplied [14C]stachyose and [14C]sucrose into leaf discs. Following short-term pulse-chase experiments with 14CO2 (15s pulse, 5 s-20 min chase), sucrose, mannitol and galactinol were rapidly labelled within the first 2 min following the pulse. Stachyose and raffinose, on the other hand, were not appreciably labelled until 10 min after the pulse. Taken together, the data indicate that phloem loading of raffinose oligosaccharides or of mannitol may not require an apoplastic step. Additionally, it appears that there may be a spatial separation of the synthesis of these sugars within the leaf, with mannitol synthesis occurring within the photosynthetic mesophyll tissues and raffinose oligosaccharide synthesis occurring closer to, and probably within, the minor veins.

Nutrient composition, protein quality and antinutritional factors of some varieties of dry beans ( Phaseolus vulgaris) grown in Burundi

Four varieties of dry beans ( Phaseolus vulgaris) grown in four different areas of Burundi were analyzed for moisture, protein, fat, ash, mineral (potassium, calcium, magnesium, iron, copper, zinc, and phosphorus), essential amino acids (isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine) and antinutritional factors (raffinose oligosaccharide, trypsin inhibitor, hemagglutinin, tannin and phytic acid) contents. In-vitro digestibility of dry bean proteins was also evaluated. The concentrations observed for the proximate composition, protein quality, and antinutritional factors varied significantly ( P < 0·05) among the 13 ‘variety-locality’ combinations investigated in this study. Mean values for moisture, protein, fat, ash, and carbohydrate were 9·19%, 22·26%, 1·01%, 4·47%, and 72·33% respectively. They were 525, 55·2, 38·2, 7·63, 0·92, 7·33 and 456 mg/100 g for potassium, calcium, magnesium, iron, copper, zinc and phosphorus, respectively; and 7·35, 14·49, 13·49, 1·59, 9·85, 9·08 and 9·11 mg/g for isoleucine, leucine, lysine, methionine, phenylalanine, threonine and valine, respectively. Finally values for raffinose, stachyose, trypsin inhibitor, hemagglutinin, tannin and phytic acid were respectively 2·20 mg/g, 18·40 mg/g, 15·02 TUI × 10 −3/g, 2·15 HU × 10 −3/mg, 14·99 mg catechin equivalent/g and 16·50 mg/g. The levels obtained for different nutrients (except for some minerals) and antinutrients were in agreement with those found in dry beans in other areas of the world. Protein digestibility ranged from 67·47% (Calima variety from Kirimiro) to 71·99% (A410 variety from Imbo). These values were comparable to those obtained in other countries. Statistical analyses of data confirmed that nutrient content, protein digestibility and undesirable factors were influenced by both variety and locality. Relationships between some dry bean nutrients and between antinutritional factors and protein digestibility were also observed.

Nutritional characteristics of soybean seed flour after processing with sodium bicarbonate or trona

Flours were prepared from soybean seeds which had been blanched in water, sodium bicarbonate (0.5%) or trona (sodium sesquicarbonate; 0.5% or 1.0%), with or without prior soaking for 24h in same solutions. All treatments removed significant proportions of the undesirable oligosaccharides raffinose and stachyose, and resulted in flours of high protein digestibility (as measured in vitro). Soaking and blanching in trona resulted in a 6-7% decrease in protein content in comparison with blanching in water alone, but this was partially offset by increases in protein digestibility and available lysine. The use of 1% trona caused some darkening of the soybean seed flour. Trona is an inexpensive and familiar processing aid, readily available to rural and urban households in Nigeria. Its use in moderate quantities may be safely recommended to reduce soaking and cooking time in the production of nutritionally and organoleptically acceptable soybean products, thereby encouraging increased consumption of this valuabl...

Oligosaccharide Metabolism and Accumulation in Developing Soybean Seeds

Soluble sugar composition of developing soybean seed (Glycine max L. Merr.) vary with cultivar. This study was conducted to determine the relation between soluble sugar accumulation and associated enzyme activities in developing seed of two soybean cultivars, Williams 82 and Wolverine. Activities of sucrose synthase, invertase, galactinol synthase, and α‐galactosidase were compared to soluble sugar content (hexoses, galactinol, myo‐inositol, oligosaccharides) at seven different reproductive growth stages. A twofold higher dry matter content in mature seeds of Wolverine corresponded to a larger seed size and longer linear seed fill. Increases in sucrose synthase and alkaline invertase activities in both cultivars corresponded to accumulation of 90% total dry matter. Acid invertase activity remained below 45 nmol glucose seed−1 min−1 during seed development. Initial raffinose and stachyose formation occurred at beginning maturity in Williams 82 and full seed in Wolverine. This change in sugar partitioning during late seed development was accompanied by twofold and threefold increases in galactinol synthase activity and galactinol content, respectively, and a substantial decrease in the substrate for this enzyme, myo‐inositol. Highest levels of galactinol synthase activity at the end of linear seed fill in both cultivars, however, preceded the accumulation of the highest stachyose levels by two growth stages. Activity of α‐galactosidase increased linearly in both varieties with time. Relatively high activities of sucrose synthase and alkaline invertase during the accumulation of the majority of dry matter in both cultivars suggest that these enzymes may be associated with this process in soybean seeds. Regulation of raffinose oligosaccharide accumulation in developing soybean seeds also may depend on galactosyl transferase activity.

Content of raffinose oligosaccharides and sucrose in various plant seeds

Etude par chromatographie liquide, au cours de la dormance et de la germination, de l'evolution des oligosides dans les differentes parties anatomiques des graines de 29 especes de plantes cultivees (graines oleagineuses, legumineuses comestibles)

Monitoring of Maillard reactions in soy products.

Chemical mechanism and the analytical methods suggested in the literature for the analysis of early Maillard reactions are summarised and critically evaluated. The applicability of our method for the measurement of the Maillard reactions - the soluble raffinose oligosaccharides and the basic amino groups of proteins - are illustrated by following early Maillard reactions in differently denaturated soya bean samples. The quantitative relations of Maillard reactions taking place in natural matrices could be clarified by monitoring both of the reactants by means of methods not requiring correction factors.

Effects of Germination on Proteins, Raffinose Oligosaccharides, and Antinutritional Factors in the Great Northern Beans (Phaseolus vulgaris L.)

ABSTRACTDuring 5 day germination, the major reserve proteins of the Great Northern bean (Phaseolus vulgaris L.) were substantially hydrolyzed. In vitro trypsin digestion of the extracted proteins up to 4 hr at 37°C indicated that the major storage proteins in germinated and the ungerminated seeds, although substantially hydrolyzed, could not be completely degraded. In vitro susceptibility of the extracted proteins was improved on moist heat treatment (97°C, 30 min). A substantial increase in soluble essential amino acids occurred during germination. Highest decrease in stachyose + raffimose (76.40%) was observed at the end of the third day of germination. Trypsin, chymotrypsin, and α‐amylase inhibitory activities and phytic acid were reduced respectively by 62.9, 73.4, 67.1, and 57.8% after 5 days of germination.

Evidence for Spider Mite (Acari: Tetranychidae) Injury-Induced Leaf Water Deficits and Osmotic Adjustment in Peppermint1

Studies were conducted to examine the physiological response of peppermint, Mentha piperita L., to leaf water stress induced by feeding of the twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). Levels of mite injury were estimated by a leaf injury index developed for these studies. Symptoms of water stress in mite-injured leaves were: a significant reduction in leaf fresh weight; reduced leaf specific weight (mg of fresh weight per cm2 of leaf area); and accumulation of soluble leaf carbohydrates. Soluble leaf carbohydrates of peppermint are sucrose (ca. 50%) and the galactosyl oligosaccharides raffinose and stachyose. Leaf starch content was unaffected by injury at all injury levels. Accumulation of soluble leaf carbohydrate in water-stressed leaves may help mite-injured leaves maintain cell turgor pressure through an osmotic adjustment (osmoregulation) mechanism. A new method for in situ starch hydrolysis for quantitative analysis of leaf starch also is presented.

Changes in starch, oligosaccharides and soluble sugars in developing pod wall and seed of chickpea

The accumulation of starch in the seed of chickpea accompanied by a decline in the pod wall during the early stages of development probably indicates that seed and pod wall did not compete with each other for starch accumulation. During the early stages of maturation, the reducing and non-reducing sugars showed a decline in seeds whereas non-reducing sugars decreased in the pod wall. The accumulation of the oligosaccharides raffinose, stachyose, and some other unidentified oligosaccharides was accompanied by a decline in the mono- and disaccharides in the developing seeds.

Distribution of α-Galactosidase in Cucurbita pepo

The distribution of alpha-galactosidase (alpha-d-galactoside galactohydrolase [EC 3.2.1.22]) in Cucurbita pepo has been determined in an attempt to assess its involvement in hydrolysis of transport sugars of the raffinose oligosaccharide series ([alpha-1-6-0-galactopyranosyl](n) sucrose). Extracts prepared from leaves and petioles at different stages of development, roots, flowers, dry and germinating seeds, all contained appreciable levels of alpha-galactosidase activity. Chromatography of these extracts on DEAE-Sephadex resolved the enzyme into three active isozymic forms. These isozymes were present in all regions of the plant analyzed but their relative proportions varied between tissues and changed within leaf and petiole tissues during development and in seeds during their germination. The level of total alpha-galactosidase activity in the leaf blade measured on a fresh weight or total protein basis remained constant at all developmental stages analyzed. The occurrence of these isozymes in mature exporting leaves indicates an effective intracellular compartmentation between their location and the sites of galactosyl oligosaccharide biosynthesis, accumulation and movement in the tissue. We have used these results to comment on the transport pathway of galactosyl oligosaccharides between the phloem and surrounding tissues in this plant.

The metabolism of galactose and the raffinose oligosaccharides by germinating bambarra groundnut seeds

Stachyose is present in the highest amount in the soluble sugar fraction of dry bambarra groundnut cotyledons, followed in descending order by raffinose, sucrose and verbascose. During germination in the dark, the stachyose and raffinose content decrease rapidly, but there is little change in the relatively small amount of verbascose present. The sucrose content increases rapidly during the first two weeks and decreases thereafter. Free glucose and fructose were present in the cotyledons after the 7th day and gradually increased in amount with time of germination. Free galactose and other galactose-containing oligosaccharides were not detected in either the dry or germinated bambarra seeds. During germination, galactose was the only identifiable sugar, aside from traces of sucrose, glucose and fructose, in the extracted soluble sugar fraction in the embryonic axes of all ages when the tissue was incubated with D-[1 14C] galactose. With the cotyledons, however, most of the radioactivity was in glucose and fructose during the early period of germination and in sucrose later. A small fraction of the radioactivity was lost as CO 2.