Changes in quantities of inositol phosphates during maturation and germination of legume seeds

 The changes in the quantities of inositol phosphates during the maturation and germination of pea, faba bean and lupin seeds were determined in two consecutive (1993 and 1994) years of differing weather conditions. Irrespective of the year, all seeds accumulated predominantly inositol hexaphosphate (IP₆). The weather conditions influenced the accumulation of inositol phosphates in maturing seeds, but they did not influence the total content. Gradual degradation of inositol phosphates occurred during seed germination. After 8 days of germination, IP₆ was degraded by some 80% in peas, 78% in faba beans and 42% in lupin seeds. The enzymic hydrolysis of higher forms of inositol phosphates (IP₆ and IP₅) in germinating seeds was assumed to yield inositol tetraphosphate (IP₄) and inositol triphosphate (IP₃), because the quantities of these compounds increased during seed germination. Received: 1 August 1997 / Revised version: 13 October 1997     

Natural Fermentation of Lentils: Influence of Time,Concentration and Temperature on the Kinetics of Hydrolysis of Inositol Phosphates

Lentil (Lens culinaris var vulgaris) flour was naturally fermented for 96 h at various conditions of concentration (79, 150 and 221 g litre⁻¹) and temperature (28°C, 35°C and 42°C). The content of total inositol phosphates (IP-total) and individual inositol phosphates (hexa- (IP₆), penta- (IP₅), tetra- (IP₄) and tri- (IP₃) phosphates) were analysed to establish the changes of these compounds during natural fermentation of lentils. The preparation of the lentil suspension brought about 16–27% reduction of the total inositol phosphates. At the end of 96 h of natural fermentation maximum IP loss (70–75%) was achieved for an experiment carried out at minimum concentration. For IP₆, the largest decrease was achieved at the highest temperature, the fermentation condition that also brought about the highest IP₅ content.     

Effect of Several Germination Treatments on Phosphatases Activities and Degradation of Phytate in Faba Bean (Vicia faba L.) and Azuki Bean (Vigna angularis L.)

Abstract: Two assays were conducted to investigate the changes of faba bean (Vicia faba L.) and azuki bean (Vigna angularis L.) phosphatases (phytase [Phy] and acid phosphatase [AcPh]) and the degradation of its substrates (inositol phosphate esters) during seed germination. The 1st assay was to establish the optimal germination conditions of faba bean and azuki bean to improve the endogenous phosphatases and increase the hydrolysis of phytate and, in the second assay, to determine the different lower phosphate esters of myo‐inositol produced during the germination process. In the 1st assay, seeds were soaked for 12 and 24 h and germinated for 3 and 5 d with and without the addition of gibberellic acid (GA₃). In the second assay, seeds were soaked for 12 h and germinated for 1, 3, and 5 d with GA₃. Phy (up to 3625 and 1340 U/kg) and AcPh (up to 9456 and 2740 U/g) activities, and inositol hexaphosphate (IP6) (8.23 and 7.46 mg/g), inositol pentaphosphate (IP5) (0.55 and 0.82 mg/g), and inositol tetraphosphate (IP4) (0.26 and 0.01 mg/g) were detected in ungerminated faba bean and azuki bean, respectively. The germination process caused a significant increase of Phy and AcPh activities in faba bean (up to 147% and 210%) and azuki bean (up to 211% and 596%) and a reduction in the phytate phosphorus content (up to 81% and 63%, respectively). Phytate phosphorus content was affected only by soaking time in the case of faba bean. Finally, during the course of germination, IP6 and IP5 were rapidly degraded in faba bean (88% and 39%) and azuki bean (55% and 56%), and IP4 was only a short‐living intermediate, which was increased during hydrolysis and degraded to inositol triphosphate. In this manner we could obtain a low‐phytate, endogenous phosphatase‐rich ingredient for enhancing human nutrition.     

New functional legume foods by germination: effect on the nutritive value of beans, lentils and peas

The effect of different conditions of germination at a semi-pilot scale on the content of available soluble sugars, alpha-galactosides, vitamins B₁ and B₂, and inositol phosphates of beans, lentils and peas have been studied. Results obtained indicated that germination modified the nutritional composition of legumes depending on the type of legume and germination conditions. The storage compounds present in dry seeds (alpha-galactosides and higher forms of inositol phosphates) decreased because they were hydrolysed to glucose, fructose, IP₄ and IP₃, compounds that can serve as a source of energy for the new plant. Vitamin B₂ suffered an important increase after germination whereas vitamin B₁ did not change significantly. To achieve legume flours with enhanced nutritive value, 6 days of germination in the presence of light for beans and lentils, and in darkness for peas can be suggested.     

Changes in chemical composition of lupin seeds (Lupinus angustifolius) after selective α‐galactoside extraction

A selective procedure for the extraction of α‐galactosides has been employed in two sweet lupin seeds (Lupinus angustifolius var. Troll and var. Emir) in order to reduce flatulence‐causing factors. Different nutritional parameters (proteins, fat, ash, dietary fibre, starch, sucrose, vitamins B₁, B₂, E and C) and antinutritional factors (α‐galactosides, trypsin inhibitor activity and inositol phosphates) were studied in raw and processed seeds. The α‐galactoside content in both varieties was reduced by 87–100%. The extracted lupins seeds presented a high retention in protein and fat (109–136% and 95–104%, respectively). Sucrose and soluble dietary fibre, however, decreased significantly as a result of processing and retentions ranged between 5 and 29%. The vitamin B₁, B₂, and E contents decreased during selective extraction, the retentions being in the ranges 25–47%, 38–40%, and 48–54%, respectively, for var. Troll and Emir. However extracted lupin seeds still contained important amounts of vitamins and insoluble dietary fibre, compounds with nutritional importance. Raw and processed lupins did not contain starch. TIA and vitamin C were not detected, and total inositol phosphates were modified slightly after extraction. In conclusion, the lupin seeds obtained by the extraction of α‐galactosides can be an adequate proteic ingredient to be incorporated in functional foods. Copyright © 2005 Society of Chemical Industry     

Functional lupin seeds (Lupinus albus L. and Lupinus luteus L.) after extraction of α-galactosides

Functional lupin seeds from two different cultivars of white (Lupinus albus L.) and yellow lupin (Lupinus luteus L.) each, were obtained by extraction of α-galactosides. The effect of extraction of α-galactosides from lupin seeds on different nutritional parameters (protein, fat, ash, dietary fibre, starch, sucrose, and vitamins B₁, B₂, E and C) and antinutritional factors (α-galactosides, trypsin inhibitor activity and inositol phosphates) were studied. In lupin seeds, α-galactosides were effectively removed and processed seeds contained very low amounts of flatulence causing factors (∼0.5–1%). Protein, fat and starch contents showed high retention in processed seeds (up to ∼130%). Sucrose and soluble dietary fibre, however, decreased significantly as a result of processing and retentions ranged from 10% to 60%, depending on the variety studied. Vitamins B₁, B₂, E and C were also reduced. Trypsin inhibitor activity was detected only in yellow lupin cultivars and inositol phosphate content was modified slightly after extraction. In summary, the functional lupin seeds, with low contents of α-galactosides, are a product of nutritional importance due to their high protein content, dietary fibre and fat contents as well as acceptable levels of thiamin, riboflavin and vitamin E. They can be incorporated as a proteic source, not only in animal feeding but also in a wide range of foods.     

Effect of Germination,under Different Environmental Conditions,on Saponins,Phytic Acid and Tannins in Lentils (Lens culinaris)

The effect of germination conditions on some antinutrients of Lens culinaris var Magda 20 seeds were studied. The seeds were germinated at 20°C under variable conditions of time, water and light. Quantitative analyses of the soyasapogenols, inositol phosphates and tannins were carried out by capillary gas chromatography, high-performance liquid chromatography and spectrophotometric techniques respectively. Germinated seeds at day 6 contained higher levels soyasapogenol B than the controls, whereas in general the tannin content was reduced. Total phytic acid amounts did not decrease after 3 days of germination but was greatly reduced after 6 days. This work shows that the optimal conditions to reduce some antinutritional factors (tannins and phytic acid) in lentils were 6 days of seed germination in dark and with alternate watering. Therefore, germination conditions offer a good opportunity to improve the nutritional quality of lentils. © 1997 SCI.     

Effect of instant controlled pressure drop on the oligosaccharides, inositol phosphates, trypsin inhibitors and lectins contents of different legumes

Well-controlled technologies for seed treatment have become a necessity for the food industry. Instant controlled pressure drop treatment (DIC®) is a new and highly controlled process that combines steam pressure (up to 8 bar) with heat (up to 170 °C) for a short time (up to 3 min). The end-product is a whole seed with a porous texture. The aim of this work was to evaluate the effect of this new (DIC) process on the contents of nutritionally active factors (NAF_s) in soybean, lupin, lentil, chickpea and roasted peanut. Unprocessed (control) and processed (DIC treatment under different pressure and time conditions) ground samples were analysed for oligosaccharides, inositol phosphates, trypsin inhibitors and lectins. The effect of DIC treatment on NAF_s in legume seeds has shown that this process considerably reduces most of these components; the optimum condition for DIC treatment in all the seeds was DIC-3 (6 bar, 1 min). The main advantages of DIC are its short processing time and the possibility of treating whole seeds for industrial applications.     

Effect of ripening stage on thiamin and riboflavin levels in lupin, pea and faba bean seeds

 Seeds of yellow pea lupin (Lupinus luteus L., cv. Juno), pea (Pisum sativum L., cv. Ergo) and faba bean (Vicia faba sp. minor Harz., cv. Tibo) were harvested at different stages of seed maturity. Changes in thiamin and riboflavin levels during growth and maturation were studied. The results obtained showed that thiamin and riboflavin were found in high abundance early in the development of lupins and peas, but that their content decreased during maturation. The thiamin content of faba bean seeds increased from 33 to 47 days after flowering (DAF), then a decrease was observed until 61 DAF; however, no significant differences between these levels in faba bean seeds harvested from 61 to 89 DAF were found. The riboflavin content of faba beans increased after 54 DAF, and no significant changes were observed between then and 89 DAF. The thiamin and riboflavin contents in lupin, pea and faba bean seeds during development were fitted to modelling curves in order to predict the vitamin content at different stages of maturity. Received: 17 July 1997     

Effect of ripening stage on thiamin and riboflavin levels in lupin, pea and faba bean seeds

 Seeds of yellow pea lupin (Lupinus luteus L., cv. Juno), pea (Pisum sativum L., cv. Ergo) and faba bean (Vicia faba sp. minor Harz., cv. Tibo) were harvested at different stages of seed maturity. Changes in thiamin and riboflavin levels during growth and maturation were studied. The results obtained showed that thiamin and riboflavin were found in high abundance early in the development of lupins and peas, but that their content decreased during maturation. The thiamin content of faba bean seeds increased from 33 to 47 days after flowering (DAF), then a decrease was observed until 61 DAF; however, no significant differences between these levels in faba bean seeds harvested from 61 to 89 DAF were found. The riboflavin content of faba beans increased after 54 DAF, and no significant changes were observed between then and 89 DAF. The thiamin and riboflavin contents in lupin, pea and faba bean seeds during development were fitted to modelling curves in order to predict the vitamin content at different stages of maturity. Received: 17 July 1997     

Evolution of soluble carbohydrates during the development of pea, faba bean and lupin seeds

Seeds of pea (Pisum sativum L. cv. Ergo), faba bean (Vicia faba ssp. minor Harz., cv. Tibo) and yellow pea lupin (Lupinus luteus L. cv. Juno) were sampled at different days after flowering (DAF) and their content of soluble carbohydrates was determined: Analysis of samples showed thatmyo-inositol, fructose, glucose, galactose and sucrose were found in high abundance early in development and their content decreased gradually during maturation. -Galactosides, which includes the content of raffinose, stachyose and verbascose, started to appear later in seed development, at 37 DAF in peas, 40 DAF in faba beans and 45 DAF in lupins. Their accumulation increased considerably during seed growth, and the maximum content was obtained in mature seeds; 3.8% in peas, 4.5% in faba beans and 10.4% in lupins. Results obtained for these sugars during seed development were fitted to modelling curves in order to predict sugar content at different development stages.     

Extraction of nitrogenous constituents from Leguminosae spp. Comparison of solubility characteristics for different species and the effect of processing conditions on protein concentrate yield

Studies were made on the solubility of nitrogen from vetch (Vicia sativa), horse bean (Vicia faba minor), pea (Pisum sativum) and lupin (Lupinus albus) seeds. With a fixed meal:solvent ratio, N-solubility is greatly increased when the pH of dispersion increases, until a pH value between 6.1-7.9 is reached. Using a fixed quantity of alkali (g NaOH kg^?1 seed meal) the pH of the suspension was kept constant. This pH was lowest with lupins and highest with vetch. The same held for solubilisation of nitrogen. For all four species, there seems to be a critical quantity of alkali below which N-solubility is poor, while above it there is little improvement. With a fixed pH of dispersion, increasing meal: solvent ratio from 10 to 50% resulted in a small decrease in N-solubility. For protein concentrate production, alkali solubilised substances were separated from the residual meal by centrifugation. The oven-dried concentrates had protein contents of 63-75%. Total protein and dry matter yield of protein concentrates (both on dry seed meal basis) ranged from 10.9 to 30.9 and from 22.6 to 49.6%, respectively, depending on seed species and conditions of extraction. In all cases yield was higher in concentrates from lupin seeds. The mineral content of protein concentrates was approximately twice that of their raw material, except for sodium.     

Germination as a process to improve the antioxidant capacity of Lupinus angustifolius L. var. Zapaton

Antioxidant capacity, measured by glutathione (GSH), superoxide dismutase-like activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), trolox equivalent antioxidant capacity (TEAC) and inhibition of lipid peroxidation in unilamellar liposomes of egg yolk phosphatidylcholine (PC) has been evaluated in raw and germinated lupin seeds (Lupinus angustifolius L. var. Zapaton) for 2, 3, 4, 5, 6 and 9 days. The content of antioxidant vitamins E and C has been also studied. The tripeptide GSH kept invariable for the first 5 days of germination and suffered a decrease of 20 and 78% after 6 and 9 days, respectively. During lupin germination, SOD-like activity increased slightly whilst PRTC doubled the amount after 9 days. TEAC values changed slightly up to 5 days of germination but after 6 and 9 days a significant increase (25 and 28%, respectively) was found. The oxidation of PC was inhibited by germinated lupin extracts and 9-day germination seeds provided the highest inhibition. Furthermore, germinated lupins provided more vitamin C, vitamin E activity and polyphenols than raw seeds, and the largest amounts of these bioactive compounds were found after 6 days of germination. Therefore, germination of lupin seeds (Lupinus angustifolius L. var. Zapaton) seems to be a good process to enhance their antioxidant capacity.     

Germination alters the chemical composition and protein quality of lupin seeds

The chemical composition and protein quality of the kernels from Lupinus angustifolius seeds were compared with that for sprouts after 6 days germination. Germination resulted in an apparent increase in protein content from 395 g kg^?1 to 435 g kg^?1 DM. Fat and carbohydrate contents decreased. The oligosaccharide content of the sprouted lupin fell to a negligible level, while the phytate and alkaloid concentrations fell from 4.7 g kg^?1 to 1.6 g kg^?1 and from 0.72 g kg^?1 to 0.16 g kg^?1, respectively. The quality of lupin kernel protein was poor with a protein efficiency ratio (PER) of 1.45±0.15. Supplementation of kernel with DL-methionine (2.0 g kg^?1) increased the protein quality (PER = 2.87±0.17) to that of casein (PER = 2.86±0.18). Germination reduced protein quality (PER = 0.44±0.16), and did not improve apparent protein digestibility (APD kernel = 80.4%; APD sprout = 77.5%). Supplementation of sprout protein with DL-methionine (2.0 g kg^?1) increased the protein quality (PER = 2.57±0.20). The total sulphur-containing amino acid concentration of lupin kernel protein, 1.9 g per 16 g N was low, and decreased further to 1.3 g per 16 g N in the sprout, a drop of 32%. The results showed that germination of lupin seeds reduced the concentration of the anti-nutritive factors; however, it also reduced protein quality.     

Inositol phosphate degradation by the action of phytase enzyme in legume seeds

The kinetics of inositol phosphate degradation during the action of naturally occurring endogenous phytase for up to 90 min in pea and lentil flours has been studied, and compared with the addition of commercial phytase enzyme. In raw lentils IP₆, IP₅, IP₄ and IP₃ were present, whilst in peas only the presence of IP₆ and IP₅ was observed. Endogenous phytases were activated when legume flour was suspended in acidified water at pH 5.5 and 37 °C, and significant differences between lentils and peas were found. IP₆ suffered a sharp reduction in lentils and peas (81–91 and 73–93%, respectively), this reduction being slightly more pronounced after the addition of commercial phytase. The content of IP₅ decreased in lentils (48–69%), and increased in peas, except when commercial phytase acted for the first 30 min, after which a reduction was found (23%). The content of IP₄ generally decreased in lentils, except when endogenous phytase acted for 30 min when an increase was observed. However, in peas, IP₄ appeared in high concentrations up to 60 min by the action of both endogenous and exogenous phytases. The content of IP₃, on the other hand, did not change greatly in lentils. In peas it was not detected after the action of endogenous phytase enzyme and it appeared in a large amount after the action of commercial phytase. In order to obtain legume flour with low IP₆ and IP₅ contents and notable IP₄ and IP₃ contents, the action of naturally endogenous phytase for 30 min in lentils is recommendable, as well as the addition of commercial phytase enzyme for 60 min in peas.     

Nutrients and antinutritional factors in faba beans as affected by processing

 The influence on the nutrients content (soluble sugars, starch, dietary fibre and calcium) and antinutritional factors (α-galactosides and phytic acid) of faba beans (Vicia faba, L. major) of soaking in different solutions (distilled water, citric acid, and sodium bicarbonate solutions), cooking the presoaked seeds, dry-heating and germination have been studied. Soaking brought about a decrease in starch, sucrose, fructose, α-galactoside, dietary fibre and calcium content. Glucose was detected in soaked faba beans and soaking did not modify the phytic acid content. Cooking the presoaked faba beans produced a slight decrease in starch, and caused a general drop in α-galactosides, dietary fibre, calcium and phytic acid, with the exception of seeds presoaked in sodium bicarbonate in which cooking did not cause any appreciable changes in comparison with the unprocessed faba beans. Germination caused a sharp reduction in α-galactoside and phytic acid content after 6 days, whilst starch and dietary fibre decreased slightly. Calcium, however, enjoyed a slight increment during germination which was related to the decrease in the content of hemicellulose and phytic acid. Dry-heating caused a noticeable reduction in all the nutrients and antinutritional factors investigated. Of all the treatments studied, germination appears to be the best processing method to obtain nutritive faba bean flour, since it caused a minor decrease in starch content (15% loss), the largest α-galactoside and phytic acid removal (94% and 45%, respectively) and provided an appreciable amount of dietary fibre. Received: 21 December 1997 / Revised version: 19 February 1998     

Inositol phosphate content and trypsin inhibitor activity in ready-to-eat cruciferous sprouts

Four cruciferous seeds (small radish, radish, white mustard and rapeseed) were germinated in order to study the fate of inositol hexaphosphate (IP-6, phytic acid) and activity of trypsin inhibitor (TIA). Reduction in the content of phytic acid depended on the time of germination. After four days of germination, when sprouts were ready-to-eat, the amount of this compound was about 50% lower in three out of the four seeds evaluated. Next, a sharp reduction in phytic acid occurred during thermal treatments (pasteurization and sterilization) of germinated rapeseed and radish sprouts. In thermal processes, a decrease in inositol hexaphosphate content was accompanied by the appearance of lower forms of inositol phosphates: IP-5, IP-4 and IP-3.The analysis of TIA, in rapeseed and radish seeds, in four-day sprouts and in these sprouts after thermal treatment, showed that only thermal processes caused complete disappearance this activity.     

Effect of natural fermentation on the content of inositol phosphates in lentils

The effects of natural fermentation upon phytic acid and less phosphorylated inositol phosphates ofLens culinaris var vulgaris cultivar Magda-20 were investigated. Seven fermentation runs were made following a 2² complete factorial design with three replicated centre points to study the effect of different conditions of temperature (28, 35 and 42°C) and broth concentration (79, 150 and 221 g/l). Samples were taken for each of them at daily intervals (0, 24, 48, 72 and 96 h). The pH value declined sharply in the first 24 h of fermentation, becoming stabilized from this time. The relation between lactic acid and titratable acidity presented important differences between the different fermentations, ranging from 30–80%. Phytic acid (IP ₆), inositol pentakis (IP ₅), tetrakis (IP ₄) and tris-(IP ₃) phosphates were quantitatively determined. The content of total inositol phosphates showed a maximum reduction of 63% at 72 h under the fermentation conditions of 42°C and 79 g/l.     

Fat and fatty acids of white lupin (Lupinus albus L.) in comparison to sesame (Sesamum indicum L.)

The study was undertaken to compare fat and fatty acid profiles in white lupin (Lupinus albus ssp. albus) and sesame (Sesamum indicum L.), representing two different families, Fabaceae and Pedaliaceae. Fat levels were 10.74% and 55.44% in seeds of white lupin and sesame, respectively. The results indicated that oleic, linolenic and arachidic acids in seed fat were higher in white lupin than in sesame cultivars. Oleic acid was the predominant fatty acid in white lupin, whereas linoleic acid was predominant in sesame. Fat content (%) was statistically significantly correlated with linoleic, linolenic and arachidic acids at the genotypic level. The fatty acid composition of white lupin is useful for human consumption. Although oil content of white lupin was lower than that of sesame, white sweet lupin could be improved.     

The effect of phytic acid on oxidative stability of raw and cooked meat

The effects of phytic acid addition (0.1, 1 and 5 mM) to pork and beef homogenates on TBARS and metmyoglobin levels in raw meat, and TBARS and heme iron contents in cooked meat during 3 days of storage at 4 °C were investigated. Also, the role of inositol as a potential synergist of IP₆ (phytic acid) was examined. IP₆ effectively decreased the TBARS accumulation in raw and cooked meat homogenates. The metmyoglobin formation was inhibited in raw beef by phytic acid in a dose-dependent manner. The effect of IP₆ was more pronounced in cooked meat than in raw and in cooked beef homogenates more than pork. Inositol did not enhance antioxidant action of phytic acid in minced meat.