Physico-Chemical Characteristics of Khesari Dhal (Lathyrus sativus): Changes in α-Galactosides,Monosaccharides and Disaccharides during Food Processing

Lathyrus sativus , also known as Khesari dhal in India, is a good source of protein and also desirable from the agricultural point of view as it is drought-resistant and can be grown with low agriculture inputs of fertiliser, irrigation and pesticide. In common with other legume seeds, Lathyrus contains the flatulence causing α-galactosides. This is a study of the physico-chemical properties of four selected Lathyrus sativus lines (Red flower, a high β-ODAP line; EC-242692, a medium β-ODAP line; L-1276, a low β-ODAP line; and Pusa-24, a very low β-ODAP line) to determine the most effective way of removing the flatulence factors by using simple processes such as soaking, dehusking and germination. Physico-chemical properties like swelling and hydration capacity of Lathyrus seeds were higher than other legumes, which suggests higher consumer preference in terms of cooking time and fuel consumption over other beans. Germination was the most effective procedure for removing α-galatosides (85–92%). Dehusking of pre-soaked seeds also achieved a high α-galactoside reduction (61–80%). Soaking in alkaline medium and thereafter boiling for 9 min brought about a lower reduction (33–74%), but these losses were still higher than those obtained by soaking in drinking water or soaking in boiled water for 2 h (11–49% and 7–44%, respectively).     

Solid-state fermentation of Lathyrus sativus seeds using Aspergillus oryzae and Rhizopus oligosporus sp T-3 to eliminate the neurotoxin β-ODAP without loss of nutritional value

The presence of a neurotoxic amino acid, 3-N-oxalyl-L-2,3-diaminopropanoic acid (β-ODAP), in the seeds of grass pea (Lathyrus sativus) is the cause of an irreversible spastic paraparesis, neurolathyrism, after over-consumption. This is a negative factor in an otherwise very tasty, nutritious, easily cultivated and hardy food crop in Asia and Africa. The reduction of β-ODAP to less than 10% of the original content has been reached by fermenting L sativas Jamalpur seeds with Aspergillus oryzae NRRL 1988 for 48 h, followed by fermentation with Rhizopus oligosporus sp T-3 for 48 h. Other nutritional qualities were also improved in the fermented seed meal: increased content of protein, higher amino acid scores for sulphur-containing and aromatic amino acids, better resistance to high temperature and to oxidation, and drastic reduction of the flatulence factors.     

β-GLUCANASE: THE SUCCESSFUL APPLICATION OF GENETIC ENGINEERING

The now well established principles of genetic engineering are described in relation to the solution of problems associated with β-glucans in beer. The application of these techniques has enabled the isolation of a Bacillus subtilis endo-1, 3–1, 4-β-D-glucanase gene which expresses a biologically active enzyme in yeast.^15,16 Although this enzyme is capable of hydrolysing beer β-glucans during fermentation, thereby enhancing beer filtration, insufficient β-glucanase is produced in yeast to enable successful commercial implementation. The requirements for the efficient production of β-glucanase in genetically manipulated brewing yeast are described.     

Effect of processing on the content of β-N-oxalyl-α, β-diaminopropionic acid (gb-ODAP) in grass pea (Lathyrus sativus) seeds and flour as determined by flow injection analysis

The effect of cooking, roasting, autoclaving and fermentation on the content of β-ODAP in the whole seeds and flour of grass pea (Lathyrus sativus) were studied at different levels of temperature, time, pH, degree of soaking and moisture content. The method of determination used was flow injection analysis, with immobilised glutamate oxidase and horseradish peroxidase. The whole seeds flour was found to contain about 922 mg 100g⁻¹ β-ODAP in dry weight basis. The reduction of β-ODAP content, in samples which were cooked for 60min and roasted (150 °C for 60min) was 57% and 82%, respectively. The content of β-ODAP in dry seeds autoclaved for 30 min also showed a significant (p = 0.05) reduction by 39%, as compared to that of raw whole seeds. Similarly, by cooking of presoaked seeds the content of β-ODAP was reduced by up to 67%. Neither the back-slopped fermentation process nor the spontaneous fermentation were effective in reducing the content of β-ODAP. Whereas roasting and autoclaving of the milled samples caused significant (p = 0.05) reduction in the content of β-ODAP up to 30% and 50%, respectively, compared to that of raw whole seeds.     

IMPROVED EXTRACTION AND ASSAY OF β-AMYLASE BROM BARLEY AND MALT

β-Amylase was extracted from barley or malt using four physical techniques to break up grists which had been prepared using a Moulinex coffee grinder. Grinding with a Polytron homogeniser apparently completely disrupted all cells, as determined by transmission electron microscopy, and increased the efficiency of extraction of β-amylase from barley by more than 30%. The other treatments tested were without value . The β-amylase activity in extracts of barley or malt was assayed by measuring the production of reducing sugars from reduced soluble starch, using a PAHBAH reagent. α-Amylase, which interferes with the quantitation of β-amylase in extracts of malt, was not totally inactivated by the chelating buffer used for enzyme extraction or by several other chelating agents. α-Amylase activity was quantified specifically using Phadebas. Using purified α-amylase a calibration was developed which related activity, as determined using Phadebas, to reducing power units. Thus the α-amylase activity present in an extract containing β-amylase could be determined using Phadebas and the reducing power equivalent activity subtracted from the total “apparent” activity to give the actual β-amylase activity. α-Glucosidase and limit dextrinase activities are believed to be too low to have a significant effect on the apparent β-amylase . The soluble and bound β-amylase activities were measured in samples taken from micromalting barley (Alexis). Dry weight losses increased to over 10% after 8 days germination. Antibiotics, applied during steeping, were used to control microbes in one experiment. However, their use checked germination and reduced malting losses to 8.4% in 8 days germination. The soluble enzyme present in extracts from steeped barley and early stages of germination was activated (20–40%) by additions of the reducing agent DTT .     

Screening grape seeds recovered from winemaking by‐products as sources of reducing agents and mammalian α‐glucosidase and α‐amylase inhibitors

Grape seeds collected from vinification of various grape varieties were extracted by supercritical CO₂ for oil recovery. The defatted residues thus obtained were considered as a re‐utilisable co‐product and assessed for phenolic content, reducing capacity and inhibitory activities against mammalian α‐amylase and α‐glucosidase enzymes. Supercritical CO₂ treatment led to higher recovery of anthocyanins. Reducing capacity of phenolic extracts reached up to ~2200 mmolFe(II) kg^?1, much higher than that of various natural phenolic sources. The anthocyanin‐rich extracts showed the highest inhibitory effectiveness towards α‐glucosidase (I₅₀ value equal to ~40 μg gallic acid equivalents (GAE)/mL ~ half than acarbose). Inhibitory effectiveness towards α‐amylase activity was similar among grape varieties, with I₅₀ values comparable to that of acarbose and correlated with proanthocyanidin contents. These results could pave the way for an efficient processing of grapes, including cascade processes, namely: winemaking, oil extraction from recovered grape seeds and phenolic extraction from defatted grape seeds as potential cost‐effective nutraceuticals.     

ENZYMIC QUANTIFICATION OF (1→3) (1→4)-β-D-GLUCAN IN BARLEY AND MALT

A simple and quantitative method for the determination of (1→3) (1→4)-β-D-glucan in barley flour and malt is described. The method allows direct analysis of β-glucan in flour and malt slurries. Mixed-linkage β-glucan is specifically depolymerized with a highly purified (1→3) (1→4)-β-D-glucanase (lichenase), from Bacillus subtilis, to tri-, tetra- and higher degree of polymerization (d.p.) oligosaccharides. These oligosaccharides are then specifically and quantitatively hydrolysed to glucose using purified β-D-glucosidase. The glucose is then specifically determined using glucose oxidase/peroxidase reagent. Since barley flours contain only low levels of glucose, and maltosaccharides do not interfere with the assay, removal of low d.p. sugars is not necessary. Blank values are determined for each sample allowing the direct measurement of β-glucan in maltsamples.α-Amylasedoes not interfere with the assay. The method issuitable for the routineanalysis of β-glucan in barley samples derived from breeding programs; 50 samples can be analysed by a single operator in a day. Evaluation of the technique on different days has indicated a mean standard error of 0–1 for barley flour samples containing 3–8 and 4–6% (w/w) β-glucan content.     

EFFECT OF PROCESSING ON NUTRITIONAL AND ANTINUTRITIONAL FACTORS OF MOONGBEAN CULTIVARS

Ordinary cooking of presoaked and unsoaked seeds of six cultivars of moongbean (Vigna radiata L.) lowered phytic acid, polyphenols, trypsin inhibitors and improved in vitro digestibility of protein and starch, and availability of Ca, Fe and Zn when compared to unprocessed seeds. However, the effect was more pronounced on ordinary cooking of presoaked seeds followed by cooking of unsoaked seeds and presoaked seeds alone. A significant increase in vitro digestibility of protein (34–48%), and starch (31–62%) may be ascribed to the reduced content of phytic acid (45–65%) polyphenols (34–70%) and trypsin inhibitor activity (42–55%) after ordinary cooking of presoaked seeds of moongbean cultivars. Therefore, ordinary cooking of soaked seeds was the most effective treatment in improving the nutritional quality of moongbean.     

TOTAL β-GLUCAN CONTENT OF SOME BARLEY CULTIVARS

The use of cellulase preparations from Trichoderma reesii for measuring the total β-glucan content of barley was examined. The activities of amyloglucosidase and β-glucanase in the cellulase varied considerably between batches, and different heat treatments were necessary to ensure that amyloglucosidase was reduced to an insignificant level while adequate β-glucanase activity was retained. After suitable treatment the cellulase was used to study variation of total β-glucan concentration in some barley cultivars. Significant varietal variation was found in the fifteen genotypes examined. These had β-glucan concentrations in the range 2.7% to 5.2% dry weight.     

In vitro inhibitory effects of cranberry bean (Phaseolus vulgaris L.) extracts on aldose reductase, α‐glucosidase and α‐amylase

The in vitro inhibitory activities of different seed extracts prepared from cranberry bean mutant SA‐05 and its wild‐type variety Hwachia against aldose reductase, α‐glucosidase and α‐amylase were examined. The results indicated that the polyphenolics‐rich extracts obtained using 800 g kg^?1 methanol and 500 g kg^?1 ethanol demonstrated inhibitory activities against aldose reductase (IC₅₀ of 0.36–0.46 mg mL^?1) and α‐glucosidase (IC₅₀ of 1.32–1.94 mg mL^?1). The 500 g kg^?1 ethanol extracts also showed α‐amylase inhibitory activities (IC₅₀ of 70.11–80.22 μg mL^?1). Subsequent extracts, prepared further with NaCl and H₂O from precipitates of 800 g kg^?1 methanol or 500 g kg^?1 ethanol extracts, exhibited potent α‐amylase inhibitory activities (IC₅₀ of 17.68–38.68 μg mL^?1). A combination of 500 g kg^?1 ethanol extraction plus a subsequent H₂O extraction produced highest polyphenolics and α‐amylase inhibitors. The SA‐05 α‐amylase inhibitor extracts showed greater inhibitory activities than that of Hwachia. Thus, cranberry bean mutant SA‐05 is an advantageous choice for producing anti‐hyperglycaemic compounds.     

ESTIMATION OF α- AND β-ACIDS IN HOP EXTRACTS BY HPLC

A procedure relying on high performance liquid chromatography for the estimation of α-acids and β-acids in hop extracts has been collaboratively tested by members of a Sub-Committee of the Institute of Brewing Analysis Committee and is recommended for use. No significant differences were found between precision values obtained using peak height and peak area measurements. For α-acids, the mean repeatability (r₉₅) and reproducibility (R₉₅) values were 1-3 and 2-4% m/m respectively over the range 41–62% m/m. For β-acids they were 0-9 and 2-0% m/m respectively over the range 11 to 35% m/m. The precision values were judged to be independent of concentration.     

Effect of Mashing Temperatures and Endo-β-Glucanase on β-Glucan Content of Malt Worts

Similar basal levels of β-D-glucans were released into worts produced at 45°C from enzymically active or inactivated flours of milled malts. In contrast, significantly higher levels of β-D-glucans were found in worts derived from either enzymically active or inactivated malt flours mashed at 65°C. In general, mashing temperature may play a more important role in releasing β-D-glucans during mashing than enzymes described as β-glucan-releasing. In this context, the physical release of β-D-glucan during mashing should be separated from the enzymic release and degradation of β-D-glucan which occur during malting.     

KINETICS OF β-GLUCAN DEGRADATION IN WORT BY EXOGENOUS β-GLUCANASES TREATMENT

Three commercial β-glucanases, one bacterial (Cereflo 200L from Novo) and two fungal (Biobeta from Gist-Brocades and Filtrase from Biocon) have been studied with regard to the hydrolysis of β-glucan in sweet and hopped wort. At temperatures below 70°C these processes follow first order kinetics with rate constants being directly proportional to the enzyme concentrations. The rate constant for bacterial β-glucanase Cereflo 200L shows a negative dependence on temperature but positive with wort pH, whereas the reverse is the case for the two fungal β-glucanases. Within the ranges of pH and temperature tested the bacterial β-glucanase has 2–5 times the activity of the fungal ones. No evidence for synergic or competitive effects between bacterial and fungal β-glucanases have been found.     

α‐Glucosidase and α‐amylase inhibitory activities of phloroglucinal derivatives from edible marine brown alga,Ecklonia cava

BACKGROUND: Diabetes mellitus (DM) is a chronic metabolic disorder characterized by defects in insulin secretion and action, which can lead to damaged blood vessels and nerves. With respect to effective therapeutic approaches to treatment of DM, much effort has being made to investigate potential inhibitors against α‐glucosidase and α‐amylase from natural products. The edible marine brown alga Ecklonia cava has been reported to possess various interesting bioactivities, which are studied here. RESULTS: In this study, five phloroglucinal derivatives were isolated from Ecklonia cava: fucodiphloroethol G ( 1 ), dieckol ( 2 ), 6,6′‐bieckol ( 3 ), 7‐phloroeckol ( 4 ) and phlorofucofuroeckol A ( 5 ); compounds 1, 3 and 4 were obtained from this genus for the first time and with higher yield. The structural elucidation of these derivatives was completely assigned by comprehensive analysis of nuclear magnetic spectral data. The anti‐diabetic activities of these derivatives were also assessed using an enzymatic inhibitory assay against rat intestinal α‐glucosidase and porcine pancreatic α‐amylase. Most of these phlorotannins showed significant inhibitory activities in a dose‐dependent manner, responding to both enzymes, especially compound 2 , with the lowest IC₅₀ values at 10.8 µmol L^?1 (α‐glucosidase) and 124.9 µmol L^?1 (α‐amylase), respectively. Further study of compound 2 revealed a non‐competitive inhibitory activity against α‐glucosidase using Lineweaver‐Burk plots. CONCLUSION: These results suggested that Ecklonia cava can be used for nutritious, nutraceutical and functional foods in diabetes as well as for related symptoms. Copyright © 2009 Society of Chemical Industry     

SUBSTRATE SPECIFICITY AND NATURE OF ACTION OF BARLEY β-GLUCAN SOLUBILASE*

Barley β/glucan solubilase was shown to be active, to differing extends, towards hot water (65°C) soluble β-glucan, CM-cellulose and cellodextrins (DP 2–8). However, the enzyme did not affect the viscosity of CM-pachyman or appear to solubilise cotton cellulose. When β/glucan was treated with lichenase a mixture of small molecular weight products was obtained including a DP 9 dextrin. This dextrin was not obtained when the β-glucan was treated with β-glucan solubilase prior to hydrolysis by lichenase. It has been concluded, therefore, that this β-glucan solubilase is an endo-type glucanase, which appears to attack the small proportion of long blocks of (1→4)-β-linked glucosyl residues reputed to be present in barley β-glucan.     

THE DEGRADATION OF β-GLUCAN DURING MALTING AND MASHING: THE ROLE OF β-GLUCANASE

Significant β-glucanolysis takes place during mashing and is catalysed by a β-glucanase which is specific to mixed-linkage β-glucans. The enzyme develops during the germination of barley, but is rapidly and extensively destroyed in kilning. Partially-purified preparations of β-glucanase are protected from denaturation by heat if their solutions are adjusted to pH 4 or if bovine serum albumin is added. However the most effective stabiliser of the enzyme is reduced glutathione. Oligosaccharides containing three and four glucosyl units are produced by the action of β-glucanase and they are further converted during malting and mashing by a different enzyme(s) to disaccharides and glucose.     

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     

The Formation of β‐Damascenone in Sweet Potato Shochu

The formation of β‐damascenone during shochu manufacture was investigated by quantifying β‐damascenone at each stage of manufacturing. Steamed sweet potato has a low level of free β‐damascenone (0.02–0.1 μg/g). During fermentation, β‐damascenone was produced in small quantities that were degraded by yeast. Thus, the second mash accumulates little free β‐damascenone (approximately 17 μg/L). The concentration profile in the fractionated distillate showed that β‐damascenone was produced during heating. Most β‐damascenone in shochu was formed during distillation, not during steam heating and fermentation. It is suggested that the level of β‐damascenone in shochu could be increased by reducing the pH of the second mash and prolonging the distillation period. Sweet potato cultivars differed in total free and hydrolyzed β‐damascenone content and there was a strong association between each cultivar and its shochu β‐damascenone content. The selection of the sweet potato cultivar is important for determining the quantity of β‐damascenone in a shochu brew.     

LEVELS OF ALKALI-SOLUBLE β-D-GLUCANS IN CEREAL GRAINS

Extraction of β-D-glucans (primarily from barley grains) using sodium hydroxide solution, gave similar results to those obtained using hydrazine extraction, suggesting that the alkali is as effective as hydrazine for the extraction of β-D-glucans. Commercial growth conditions in different European countries caused limited shifts in the β-D-glucan levels of malting barley samples. Sorghum and wheat grains contained significantly less β-D-glucans than barley. Potassium bromate solution inhibited initial breakdown of β-D-glucans during the malting of barley grains. The breakdown of β-D-glucan in Sonja barley was much slower than in Golden Promise barley, during malting.     

PENICILLIUM FUNICULOSUM AS A SOURCE OF β-GLUCANASE FOR THE ESTIMATION OF BARLEY β-GLUCAN

The β-glucanase in commercial |cellulases prepared from Penicillium funiculosum is more stable to heating than is the equivalent enzyme system from Trichoderma reesei. Consequently the heat-labile amyloglucosidase can be destroyed more reliably in Penicillium cellulase, and it follows that this enzyme is to be preferred for use in the enzymic method for measuring β-glucan.