Acceptability,storage stability and costing of α‐amylase‐treated maize–beans–groundnuts–bambaranuts complementary blend

The effects of α‐amylase treatment on physical properties, acceptability to mothers, and cost of roasted and extruded maize–beans–groundnuts–bambaranuts complementary porridge recipes were assessed prior to their industrial production. Storage stability of the extruded α‐amylase‐treated fortified blend was assessed at 2 weeks and 6 months by sensory evaluation, peroxide value, water activity and microbiological load. The use of α‐amylase at 0.04% w/w enhanced porridge acceptability and resulted in 88% and 122% increase in flour concentration for roasted and extrusion‐cooked porridge flour, respectively, while maintaining porridge viscosity at 1000‐fold lower than that of traditionally used porridges. The extrusion cooked blend was stable for 6 months. α‐Amylase application increased the unit cost of the developed blend by only 1.4%. The total cost was less than US $ 2 kg^?1, half the minimum price of commercially available complementary foods. Further work on marketing and the efficacy of this inexpensive food on growth of infants is warranted. Copyright © 2007 Society of Chemical Industry     

Chemical,biological and sensory evaluation of pasta products supplemented with α‐galactoside‐free lupin flours

α‐Galactoside‐free lupin flour has been used to supplement durum wheat semolina flour in order to increase the nutritive value of pasta products. Supplemented pasta products had a shorter cooking time, higher cooking water absorption, cooking loss and protein loss in water than control pasta prepared with only semolina. Sensory evaluation of cooked pastas showed that products supplemented with 80 g kg^?1 of α‐galactoside‐free Lupinus angustifolius var. Emir flour or with 100 g kg^?1 of α‐galactoside‐free Lupinus angustifolius var. Troll flour showed the same acceptability by panellists as the semolina pasta. These levels of supplementation were selected for further studies. The cooked α‐galactoside‐free lupin/semolina pastas showed higher amounts of protein, dietary fibre, calcium, phosphorus, magnesium, zinc and antioxidant capacity than control pasta and a reasonable level of vitamin B₁, vitamin B₂ and vitamin E. Biological assessment of cooked pastas indicated that the true protein digestibility did not change after the fortification of semolina but protein efficiency ratio increased sharply in the pasta supplemented with α‐galactoside‐free lupin flours (2.07 and 1.92 for Emir and Troll lupin varieties, respectively) in comparison with the control pasta (1.11). It is concluded that the α‐galactoside‐free lupin flours are an adequate ingredient to improve the nutritional quality of pasta products without adding flatulent oligosaccharides. Copyright © 2006 Society of Chemical Industry     

Effects of germination on the composition and nutritive value of proteins in Pisum sativum,L

The effects of germination for 2, 4 or 6 d, with and without light, on the proteolytic activity, the contents of soluble protein and non protein nitrogen, and the amount of available starch of Pisum sativum, L, as well as their nutritive utilisation by growing rats were studied. Food intake increased significantly when the peas were germinated for 2 or 4 d. This improvement was correlated with the reduction of factors responsible for flatulence. Digestive utilization of nitrogen was similar (among all the groups fed germinated-pea flour) to raw-pea flour. The values for nitrogen balance, percentage of retained to absorbed nitrogen, protein efficiency ratio, and index of available carbohydrates were significantly higher among the animals that consumed peas allowed to germinate for 2 or 4 d than among the animals given the raw-pea or 6-day-germinated pea diets. We conclude that germination of peas for 2 d would be sufficient to significantly improve the palatability and nutritive utilisation of protein and carbohydrates from Pisum sativum, L. The presence or absence of light during the germination process did not affect the results achieved.     

Investigation of the interaction between (−)‐epigallocatechin‐3‐gallate with trypsin and α‐chymotrypsin

Tea polyphenol (TP) inhibits digestive enzymes and reduces food digestibility. To explore the interaction between TP with digestive enzymes, bindings of ‐epigallocatechin‐3‐gallate (EGCG) to trypsin and α‐chymotrypsin were studied in detail using fluorescence, resonance light‐scattering, circular dichroism, fourier transform infrared spectroscopy methods and protein‐ligand docking. The binding parameters were calculated according to Stern–Volmer equation, and the thermodynamic parameters were determined by the van't Hoff equation. The results indicated that EGCG was capable of binding trypsin and α‐chymotrypsin with high affinity, resulting in a change of native conformation of these enzymes. EGCG had a greater influence on the structure of α‐chymotrypsin than trypsin. This study can be used to explain the binding interaction mechanism between TP and digestive enzymes.     

Phytate content and phytate degradation by endogenous phytase in pea (Pisum sativum)

In order to rapidly reduce the content of inositol tri–hexaphosphates in pea flour by action of the endogenous phytase, raw materials as well as incubation conditions have been evaluated. The phytate (inositol hexaphosphate) content was analysed in 27 pea varieties; the influence of storage time and the difference in phytate content between the germ and the cotyledon were determined. Furthermore, degradation of inositol phosphates by the endogenous phytase enzyme was studied in pea flour, germ and cotyledon. To find the maximum phytate degradation, the effects of temperature and pH during pea flour incubation were investigated. The most efficient phytate degradation in pea flour incubation was achieved at pH 7.5 and 45 °C. At this condition an almost complete degradation of phytate and a 66% reduction in the sum of inositol hexa‐, penta‐, tetra‐ and triphosphates were reached in 10 h. The storage time of pea seeds or removal of the germ did not have a major effect on the phytate content. Since several inositol pentaphosphate isomers were produced during phytate degradation, it can be concluded that peas contain several phytate‐degrading enzymes, or one phytate‐degrading enzyme with unspecific initial hydrolysation pattern. © 2001 Society of Chemical Industry.     

Significance of healthy viscous dietary fibres on the performance of gluten‐free rice‐based formulated breads

The impact of associated viscous dietary fibres (hydroxypropylmethylcellulose semi‐firm – SFE‐ and weak – NE‐gel forming, and barley ß‐glucan, BBG) incorporated at different amounts (1.6–7.5%, flour basis) into gluten‐free rice‐based dough formulations on the breadmaking performance and staling behaviour of hydrated (70–110%, flour basis) fibre‐flour composite blends has been investigated. Single BBG addition fails to mimic gluten visco‐elasticity properly, but simultaneous incorporation of either SFE or NE contributes to bread improvement in terms of bigger volume and smoother crumb. 3.3 g of BBG (70% purity) and 104 mL of water addition to 100 g rice flour provided sensorially accepted breads (7.6/10) with a theoretical ß‐glucan content of 1.24 g per 100 g GF bread that would allow a daily ß‐glucan intake of 3 g provided a bread consumption of 240 g day^?1. Complementary tests should be carried out to know the amount and molecular weight of ß‐glucan in the final bread before assuring the nutritional benefit of this addition.     

Hydrophobicity‐modulating self‐assembled morphologies of α‐zein in aqueous ethanol

Protein fibrillation serves a broad range of biological functions from surface colonising to mechanically reinforcing structures; it is also associated with the development of neurodegenerative disorders. Although fibrillation is considered to be an inherent ability of polypeptides to form backbones, relevant studies have long concentrated on aqueous‐soluble proteins rather than on insoluble structures. In this study, the self‐assembly of hydrophobic proteins into amyloid nanofibrils was studied using α‐zein as a model protein. The self‐assembled morphologies of α‐zein were determined by hydrophobic–hydrophilic characters of both the protein and the solvent. Upon thermally incubating in aqueous ethanol, α‐zein formed amyloid nanofibrils with lower ethanol compositions and near‐neutral pHs, while acidic conditions and high ethanol compositions result in the formation of spherical aggregations. The fibrillation of α‐zein shows a great potential in serving as bio‐based gels or reinforced fillers in food, cosmetic and pharmaceutical applications.     

Mango wine aroma enhancement by pulp contact and β‐glucosidase

This study determined the influence of pulp maceration and β‐glucosidase on mango wine physico‐chemical properties and volatile profiles. The distinction in pH, sugars and organic acids among different treatments was not statistically significant. All wine samples reached around 8% (v/v) ethanol from about 16% (w/v) of sugars. The wine with pulp contact contained about ten times higher α‐terpinolene and up to three times higher acetate esters than the wine without pulp contact, but mitigated the production of medium‐chain fatty acids and relative ethyl esters by up to six times. β‐glucosidase enhanced terpenols by up to ten times and acetate esters by up to three times. Furthermore, enzyme treatment mitigated, by up to five times, the formation of medium‐chain fatty acids and ethyl esters to moderate levels. Sensory evaluation showed pulp contact, and β‐glucosidase not only improved the intensity and complexity of wine aroma but balanced odour attributes.     

A critical review of methodologies used in determination of relative bio‐availability ratio of RRR‐α‐tocopheryl acetate and all‐rac‐α‐tocopheryl acetate

Bio‐availability of different α‐tocopherol forms in livestock animals is measured by the increase in plasma or tissue concentrations of α‐tocopherol after oral administration. It is generally accepted that RRR‐α‐tocopheryl acetate (natural source vitamin E derived from vegetable oil) has a higher bio‐availability compared to all‐rac‐α‐tocopheryl acetate (synthetic vitamin E, i.e. α‐tocopherol produced by chemical synthesis). However, different bio‐availability ratios have been reported in the literature. The major reason for conflicting results in literature studies was the inability to separate the proportion of α‐tocopherol originating from test materials, from the proportion of α‐tocopherol originating from basal dietary ingredients and pre‐feeding. This causes significant variability. For bio‐availability determination, a baseline or control treatment is essential. The estimation of bio‐availability without correction for basal vitamin E status will lead to incorrect interpretation of the results. When using proper methodologies, it is possible to correct for the impact of α‐tocopherol intake from basal ingredients and α‐tocopherol originating from pre‐feeding, therefore yielding results reflecting the true relative bio‐availability of different α‐tocopherol substances. When reviewing literature data a critical evaluation of the method used in determination of relative bio‐availability is recommended. Copyright © 2010 Society of Chemical Industry     

Validation of analytical method for α‐dicarbonyl compounds using gas chromatography–nitrogen phosphorous detector and their levels in alcoholic beverages

This study aimed to establish an analytical method for α‐dicarbonyl compounds (α‐DCs) including glyoxal, methylglyoxal and diacetyl, to determine the content of α‐DCs in 101 various alcoholic beverages using gas chromatography–nitrogen phosphorous detector (GC‐NPD) and to perform exposure assessment. The limit of detection and limit of quantification for α‐DCs were 0.05–0.22 and 0.15–0.70 μg g^?1, respectively. The accuracy and precision were validated in five matrices. The raspberry fruit wine had the highest value at 139.74 μg g^?1 total α‐DCs. The lowest α‐DC concentration among the beverages was detected in rice wine (Makgeolli) at 1.59 μg g^?1. The levels of α‐DCs in various samples were detected as follows: 1.59–56.68 μg g^?1 in rice wine (Makgeolli), 2.73–16.77 μg g^?1 in beer, 8.22–139.74 μg g^?1 in fruit wine and 8.17–91.56 μg g^?1 in rice wine (Cheongju). The estimated daily intake of α‐DCs in the intake‐only group and population group was calculated as 4.22–97.94 μg kg^?1 bw day^?1 and 0.28–7.13 μg kg^?1 bw day^?1, respectively.     

Edible foxtail millet flour stabilises and retain the in vitro activity of blueberry bioactive components

Blueberries, a functional food, are rich in bioactive polyphenols and anthocyanins. However, the shelf life is short and requires cold storage. This study provides evidence that edible foxtail millet flour (FMF) efficiently sorbs only blueberry bioactive components (polyphenols and anthocyanins) but not sugars, improves their stability and retains the activity. The concentration of blueberry polyphenols and anthocyanins sorbed to FMF ranged from 6 to 113 and 4 to 41 mg g^?1, respectively. The concentration of bioactive components in one serving of blueberries (73 g) is equivalent to those present in 1.2 g of blueberry‐enriched foxtail millet flour (BFMF). The blueberry bioactive sorbed onto FMF remained stable for at least 16 weeks at 40 °C. BFMF eluates inhibited α‐glucosidase enzyme activity and scavenged the free radicals conferring that blueberry bioactive components in BFMF retained the activity. The sorption process described here provides a practical way of creating low glycemic protein‐rich edible flour enriched with plant bioactive compounds without sugars.     

Purification and functional characterisation of an α‐l‐rhamnosidase from Penicillium citrinum MTCC‐3565

An extracellular α‐l ‐rhamnosidase from Penicillium citrinum MTCC‐3565 has purified to homogeneity from its culture filtrate using ethanol precipitation and cation‐exchange chromatography on carboxymethyl cellulose. The purified enzyme gave a single protein band corresponding to molecular mass of 45.0 kDa in SDS‐PAGE analysis showing the purity of the enzyme preparation. The native PAGE analysis showed the monomeric nature of the purified enzyme. Using p‐nitrophenyl α‐l ‐rhamnopyranoside as substrate, K_m and V_max values of the enzyme were 0.30 mm and 27.0 μm min mg^?1, respectively. The k_cat value was 20.1 s giving k_cat/K_m value of 67.0 mm s^?1 for the same substrate. The pH and temperature optima of the enzyme were 8.5 and 50 °C, respectively. The activation energy for the thermal denaturation of the enzyme was 29.9 KJ mol^?1. The α‐l ‐rhamnosidase was able to hydrolyse naringin, rutin and hesperidin and liberated l ‐rhamnose, indicating that the purified enzyme can be used for the preparation of α‐l ‐rhamnose and pharmaceutically important compounds by derhamnosylation of natural glycosides containing terminal α‐l ‐rhamnose. The α‐l ‐rhamnosidase was active at the level of ethanol concentration present in wine, indicating that it can be used for improving wine aroma.     

Biological response of broiler chickens fed peas (Pisum sativum L.) expressing the bean (Phaseolus vulgaris L.) α‐amylase inhibitor transgene

The nutritive value of transgenic peas expressing an α‐amylase inhibitor (α‐Ai1) was evaluated with broiler chickens. The effects of feeding transgenic peas on the development of visceral organs associated with digestion and nutrient absorption were also examined. The chemical composition of the conventional and the transgenic peas used in this study were similar. In the two feeding trials, that were conducted normal and transgenic peas were incorporated into a maize–soybean diet at concentrations up to 500 g kg^?1. The diets were balanced to contain similar levels of apparent metabolisable energy (AME) and amino acids. In the first trial, the birds were fed the diets from 3 to 17 days post‐hatching and with levels of transgenic peas at 250 g kg^?1 or greater there was a significant reduction in body weight but an increase in feed intake resulting in deceased feed conversion efficiency. In the second trial, in which the birds were fed diets containing 300 g kg^?1 transgenic peas until 40 days of age, growth performance was significantly reduced. It was also demonstrated that the ileal starch digestibility coefficient (0.80 vs 0.42) was significantly reduced in the birds fed transgenic peas. Determination of AME and ileal digestibility of amino acids in 5‐week‐old broilers demonstrated a significant reduction in AME (12.12 vs 5.08 MJ kg^?1 DM) in the birds fed the transgenic peas. The AME value recorded for transgenic peas reflected the lower starch digestibility of this line. Ileal digestion of protein and amino acids was unaffected by treatment. Expression of α‐Ai1 in peas did not appear to affect bird health or the utilisation of dietary protein. However, the significant reduction in ileal digestion of starch in transgenic peas does reduce the utility of this feedstuff in monogastric diets where efficient energy utilisation is required. Copyright © 2006 Society of Chemical Industry     

α‐l‐Rhamnosidase from Aspergillus flavus MTCC‐9606 isolated from lemon fruit peel

An α‐l ‐rhamnosidase producing fungal strain has been isolated from decaying lemon fruit. The fungal strain has been identified as Aspergillus flavus. The α‐l ‐rhamnosidase has been purified from the culture filtrate of the fungal strain using ultra filtration and cation exchange chromatography on carboxy methyl (CM) cellulose. The molecular mass of the purified enzyme determined by SDS–PAGE analysis was 41 kDa. The K_m values of the enzyme using p‐nitrophenyl‐α‐l ‐rhamnopyranoside and naringin as the substrates were 1.89 and 1.6 mm respectively. The pH and temperature optima of the enzyme were 11.0 and 50 °C respectively. The effects of various chemical species present in grape fruit juice and wine on the activity of the enzyme have been determined.     

The effect of pea (Pisum sativum L.)‐originated asparaginase on acrylamide formation in certain bread types

The aim of this study was to investigate the effect of pea (Pisum sativum L.)‐originated asparaginase on acrylamide formation in white wheat, wheat bran and whole‐grain wheat breads. Two‐day germinated pea flour was used at 0%, 1%, 3% and 5% levels for each bread type. Acrylamide analysis was performed with liquid chromatography–mass spectrometry. Besides, colour and sensory properties of the breads were evaluated to search out the effects of pea flour substitution on the consumer acceptance. Reduction of acrylamide in white wheat bread was not found significant and addition of pea flour decreased the acceptance. However, it was found that acrylamide level can be reduced by 57% and 68% with addition of 5% pea flour in wheat bran and whole‐grain breads, respectively, without any negative impact on colour and sensory properties.     

Purification and characterisation of a novel α‐L‐rhamnosidase exhibiting transglycosylating activity from Aspergillus oryzae

A novel α‐L‐rhamnosidase was isolated and purified from Aspergillus oryzae NL‐1. The enzyme was purified 13.2‐fold by ultrafiltration, ion exchange and gel filtration chromatography with an overall recovery of 6.4% and specific activity of 224.4 U/mg, and the molecular mass of its subunit was approximately 75 kDa. Its optimal temperature and pH were 65 °C and 4.5, respectively. The enzyme was stable in the pH range 3.5–7.0, and it showed good thermostability at higher temperatures. The K_M, kcat and kcat/K_M values were 5.2 mm , 1624 s^?1 and 312 s^?1 mm ^?1 using pNPR as substrates, respectively. Moreover, the enzyme exhibited transglycosylating activity, which could synthesise rhamnosyl mannitol through the reactions of transglycosylation with inexpensive rhamnose as the glycosyl donor. Our findings indicate that the enzyme has potential value for glycoside synthesis in the food industry.     

The Advantages of Using Natural Substrate‐Based Methods in Assessing the Roles and Synergistic and Competitive Interactions of Barley Malt Starch‐Degrading Enzymes

Existing methods of assay of malt starch‐degrading enzymes were critically appraised. New methods based on natural substrates, namely starch and its natural intermediate‐derivative, were developed for all the enzymes, except limit dextrinase for which pullulan was used. Thermostability, optimal temperatures and pHs were established. α‐Amylase and limit dextrinase were the most thermostable and β‐amylase, α‐glucosidase and maltase were the least stable while diastase occupied an intermediate position. The optimal temperatures were congruent with thermostability, β‐ amylase having the lowest (50°C) and α‐amylase the highest (65°C) with the remaining enzymes, including diastase, falling in between. In contrast, α‐amylase has the lowest optimal pH (pH 4.5) and β amylase the highest (pH 5.5) while the others have pHs in between the two values. The roles of the enzymes were evaluated taking into account the level of activity, thermostability, optimum pH, the nature of the product(s), and the relevance to brewing. β‐Amylase production of maltose was synergistically enhanced, mostly by α‐amylase but also limit dextrinase. α‐Glucosidase and maltase are unimportant for brewing, because of their low activity and the negative impact on β‐amylase activity and the negative effect of glucose on maltose uptake by yeast. The starch‐degrading enzymes (diastase) in a gram of malt were able to degrade more than 8 g boiled starch into reducing sugars in 10 min at 65°C. The latter, suggests that it will be possible to gelatinise most of the malt starch at a higher temperature and ensure its hydrolysis to fermentable sugars by mixing with smaller portions of malt and mashing at lower temperatures e.g. 50–60°C.     

Isolation of a Recombinant Bacillus sp. TS‐23 α‐Amylase by Adsorption‐Elution on Raw Starch

A Bacillus sp. TS‐23 α‐amylase produced by recombinant Escherichia coli was adsorbed onto raw starch and the adsorbed enzyme was eluted with maltose or maltodextrin in 50 mM Tris/HCl buffer (pH 8.5). The adsorption‐elution procedure resulted in a yield of 53% α‐amylase activity and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS/PAGE) analysis showed that the eluted α‐amylase had a molecular mass of approximately 64 kDa. Raw starch could be used repeatedly in the adsorption‐ elution cycle with good reproducibility. Scanning electron microscopy of the isolated corn starch exhibited a smooth appearance of the granules before adsorption and only a small change in appearance after three adsorption‐elution cycles. These results suggest that the raw starch adsorption‐elution technique has a great potential in the isolation of Bacillus sp. TS‐23 α‐amylase from the culture broth of recombinant E. coli.     

Inhibitory potential of phenylpropanoid glycosides from Ligustrum purpurascens Kudingcha against α‐glucosidase and α‐amylase in vitro

The leaves of Ligustrum purpurascens are used in a Chinese traditional tea called small‐leaved kudingcha, which is rich in phenylpropanoid glycosides (PPGs) and has many beneficial properties. Two critical exoacting glycoside hydrolase enzymes (glucosidases) involved in carbohydrate digestion are α‐glucosidase and α‐amylase. We investigated the properties of PPGs from L. purpurascens for inhibiting α‐amylase and α‐glucosidase activity in vitro and found IC₅₀ values of 1.02 and 0.73 mg mL^?1, respectively. The patterns of inhibiting both α‐amylase and α‐glucosidase were mixed‐inhibition type. Multispectroscopy and molecular docking studies indicated that the interaction between PPGs and α‐amylase and α‐glucosidase altered the conformation of enzymes, with binding at the site close to the active site of enzymes resulting in changed enzyme activity. Our studies may help in the further health use of small‐leaved kudingcha.