Isolation, fractionation and characterisation of proteins from Mucuna bean

The chemical composition and fractional distribution of protein isolates prepared from species of Mucuna bean were studied. Using six different extraction media, the yield of protein based on the Kjeldahl procedure varied from 8% to 34%, and the protein content varied from 75% to 95%. When the yields were high, the colour of the isolates generally tended to be dark and unsatisfactory. Hence, the use of chemical treatments and high pressure processing were explored. The solubility maxima for the protein isolates in water were found to occur at pH values of 2.0 and 11.0, while the pH corresponding to minimum solubility (i.e. isoelectric region) occurred at pH values of 4.0 and 5.0. The total essential amino acid in the isolates ranged from 495 to 557 mg g(-1) protein, which compares favourably with the recommended level for pre-school and school children. Methionine and cysteine were the limiting amino acids. A key nutritional attribute of the protein isolates was its high lysine content. The isolate can therefore complement cereal-based foods which are deficient in lysine. The proteins mainly consisted of albumins, glutelins and globulins. Prolamins were only present in trace concentration (< 0.3%). Gel filtration chromatograms of the isolates indicated the presence of major protein fractions with molecular weights of 40 and 15 kDa, while gel electrophoresis (SDS-PAGE) indicated a major broad zone with molecular weights of 36 +/- 7 and 17.3 +/- 13 kDa. (c) 2006 Elsevier Ltd. All rights reserved.

Heat-inactivated peroxidases and the role of calcium abstraction as a cause of their enhanced lipid oxidation activity: potential effects on the flavour quality of heat-processed vegetables

Cationic swede and anionic turnip peroxidases were partially purified by ion-exchange and gel-filtration chromatography, respectively. Heat treatment of these enzymes and of a commercial high purity horseradish peroxidase (HRP) caused a loss of enzyme activity and a corresponding increase in linoleic acid hydroperoxide formation activity. The hydroperoxide levels in model systems increased only in the early stages of the oxidation reaction and then declined as degradation became more significant. The presence of a dialysed blend of cooked swede markedly lowered the hydroperoxide level formed. Analysis of volatile compounds formed showed that hexanal predominated in a buffer system and in a blend of cooked turnip. In dialysed blends of cooked swede, hexanol was the primary volatile compound generated. After inactivation under mild conditions in the presence of EDTA, the peroxidases showed hydroperoxide formation activity and patterns of volatile compounds from linoleic acid that were similar to those found on heat-inactivation. This suggested that calcium abstraction from the peroxidases was critical for the enhancement of lipid oxidation activity. (C) 2002 Elsevier Science Ltd. All rights reserved.

Purification and functional characterisation of Rhinocerase, a novel serine protease from the venom of Bitis gabonica rhinoceros

Background: Serine proteases are a major component of viper venoms and are thought to disrupt several distinct elements of the blood coagulation system of envenomed victims. A detailed understanding of the functions of these enzymes is important both for acquiring a fuller understanding of the pathology of envenoming and because these venom proteins have shown potential in treating blood coagulation disorders. Methodology/Principal Findings: In this study a novel, highly abundant serine protease, which we have named rhinocerase, has been isolated and characterised from the venom of Bitis gabonica rhinoceros using liquid phase isoelectric focusing and gel filtration. Like many viper venom serine proteases, this enzyme is glycosylated; the estimated molecular mass of the native enzyme is approximately 36kDa, which reduces to 31kDa after deglycosylation. The partial amino acid sequence shows similarity to other viper venom serine proteases, but is clearly distinct from the sequence of the only other sequenced serine protease from Bitis gabonica. Other viper venom serine proteases have been shown to exert distinct biological effects, and our preliminary functional characterization of rhinocerase suggest it to be multifunctional. It is capable of degrading α and β chains of fibrinogen, dissolving plasma clots and of hydrolysing a kallikrein substrate. Conclusions/Significance: A novel multifunctional viper venom serine protease has been isolated and characterised. The activities of the enzyme are consistent with the known in vivo effects of Bitis gabonica envenoming, including bleeding disorders, clotting disorders and hypotension. This study will form the basis for future research to understand the mechanisms of serine protease action, and examine the potential for rhinocerase to be used clinically to reduce the risk of human haemostatic disorders such as heart attacks and strokes.

Permeate flux modeling of membrane distillation

This paper concerns the modeling of membrane distillation. The model developed has been used to predict permeate fluxes using different initial operating conditions. PVDF and PTFE membranes were successfully used in a flat plate module to experimentally confirm the theoretical results. The correlation between theory and experiment was close for both membranes. The PTFE membranes produced higher fluxes than PVDF. A Versapor membrane was also used for this work. This membrane is a composite, with a thin porous layer on a support layer. It was found not to be suitable for membrane distillation. A comparison of the heat flux was also carried out. Again, there was good correlation between theory and experiment

Corticotropin-releasing factor binding protein dimerizes after association with ligand.

We have suggested recently that the fall in plasma CRF-binding protein (BP) during the last few weeks of pregnancy is a direct effect of association with its ligand because of the rapid decrease in plasma BP concentration seen in normal males reaching a nadir some 15 min after a bolus injection of synthetic CRF. In the present study, we have investigated the physicochemical properties of both natural and recombinant BP by gel filtration under physiological conditions and have shown that association of human CRF to this BP results in an increase in molecular weight consistent with the formation of a dimer form of the BP ligand complex. The dimer is more stable when the interaction occurs in the presence of serum or if a peptide with a higher affinity for the BP is substituted as ligand. Experimental evidence would also suggest that the dimer BP has a higher affinity for ligand than the monomeric form. We suggest that this dimerization occurs in vivo when CRF is released into the bloodstream and provides the trigger that causes the uptake of the complex at specific receptor sites.

Adoption of green revolution services and poverty reduction in Ghana

In Sub-Saharan Africa (SSA) the technological advances of the Green Revolution (GR) have not been very successful. However, the efforts being made to re-introduce the revolution call for more socio-economic research into the adoption and the effects of the new technologies. The paper discusses an investigation on the effects of GR technology adoption on poverty among households in Ghana. Maximum likelihood estimation of a poverty model within the framework of Heckman's two stage method of correcting for sample selection was employed. Technology adoption was found to have positive effects in reducing poverty. Other factors that reduce poverty include education, credit, durable assets, living in the forest belt and in the south of the country. Technology adoption itself was also facilitated by education, credit, non-farm income and household labour supply as well as living in urban centres. Inarguably, technology adoption can be taken seriously by increasing the levels of complementary inputs such as credit, extension services and infrastructure. Above all, the fundamental problems of illiteracy, inequality and lack of effective markets must be addressed through increasing the levels of formal and non-formal education, equitable distribution of the 'national cake' and a more pragmatic management of the ongoing Structural Adjustment Programme.

The impact of climate change on the treatability of dissolved organic matter (DOM) in upland water supplies: A UK perspective

Climate change in the UK is expected to cause increases in temperatures, altered precipitation patterns and more frequent and extreme weather events. In this review we discuss climate effects on dissolved organic matter (DOM), how altered DOM and water physico-chemical properties will affect treatment processes and assess the utility of techniques used to remove DOM and monitor water quality. A critical analysis of the literature has been undertaken with a focus on catchment drivers of DOM character, removal of DOM via coagulation and the formation of disinfectant by-products (DBPs). We suggest that: (1) upland catchments recovering from acidification will continue to produce more DOM with a greater hydrophobic fraction as solubility controls decrease; (2) greater seasonality in DOM export is likely in future due to altered precipitation patterns; (3) changes in species diversity and water properties could encourage algal blooms; and (4) that land management and vegetative changes may have significant effects on DOM export and treatability but require further research. Increases in DBPs may occur where catchments have high influence from peatlands or where algal blooms become an issue. To increase resilience to variable DOM quantity and character we suggest that one or more of the following steps are undertaken at the treatment works: a) ‘enhanced coagulation’ optimised for DOM removal; b) switching from aluminium to ferric coagulants and/or incorporating coagulant aids; c) use of magnetic ion-exchange (MIEX) pre-coagulation; and d) activated carbon filtration post-coagulation. Fluorescence and UV absorbance techniques are highlighted as potential methods for low-cost, rapid on-line process optimisation to improve DOM removal and minimise DBPs.

Control of final moisture content of food products baked in continuous tunnel ovens

There are well-known difficulties in making measurements of the moisture content of baked goods (such as bread, buns, biscuits, crackers and cake) during baking or at the oven exit; in this paper several sensing methods are discussed, but none of them are able to provide direct measurement with sufficient precision. An alternative is to use indirect inferential methods. Some of these methods involve dynamic modelling, with incorporation of thermal properties and using techniques familiar in computational fluid dynamics (CFD); a method of this class that has been used for the modelling of heat and mass transfer in one direction during baking is summarized, which may be extended to model transport of moisture within the product and also within the surrounding atmosphere. The concept of injecting heat during the baking process proportional to the calculated heat load on the oven has been implemented in a control scheme based on heat balance zone by zone through a continuous baking oven, taking advantage of the high latent heat of evaporation of water. Tests on biscuit production ovens are reported, with results that support a claim that the scheme gives more reproducible water distribution in the final product than conventional closed loop control of zone ambient temperatures, thus enabling water content to be held more closely within tolerance.

Enhancing the crystallization and orientation of electrospinning poly (lactic acid) (PLLA) by combining with additives

PLLA is a thermoplastic biopolymer and can be used in industrial applications for medical and filtration applications. The brittleness of PLLA is attributed to slow crystallization rates and its glass transition temperature (Tg) is high (60 °C); for this reason, its applications are limited. The orientation, morphology, and crystal structure of the electrospun fibers was investigated by SEM, POM, DSC, FTIR, XRD, and SAXS. Combining with additives leads to a large decrease of fiber diameter, viscosity, and changes of fiber morphology and crystal structure compared to pure PLLA. DSC showed that the Tg of PLLA decreased about 15 °C and there was no change in relaxation enthalpy by the addition of plasticizer. FT-IR indicate a strong interaction between PLLA and additives; a new band appears in the PLLA blend at 1,756 cm−1 at room temperature as a crystalline band without any annealing. In addition, WAXD indicated that the intensities of the two peaks at (200/110) and (203) increased for the blend at room temperature without any annealing in comparison with PLLA; this means that PHB crystallizes in the amorphous region of PLLA. The POM experiments agree with the results from DSC, FTIR, and WAXS measurements, confirming that adding PHB results in an increase in the number of nuclei with much smaller spherulites and enhances the crystallization behavior of this material, thereby improving its potential for applications.

Replacing fat and sugar with inulin in cakes: bubble size distribution, physical and sensory properties

The replacement of fat and sugar in cakes is a challenge as they have an important effect on the structural and sensory properties. Moreover, there is the possibility to incorporate an additional value using novel replacers. In this work, inulin and oligofructose were used as fat and sugar replacers, respectively. Different combinations of replacement levels were investigated: fat replacement (0 and 50 %) and sugar replacement (0, 20, 30, 40 and 50 %). Simulated microbaking was carried out to study bubble size distribution during baking. Batter viscosity and weight loss during baking were also analysed. Cake characteristics were studied in terms of cell crumb structure, height, texture and sensory properties. Fat and sugar replacement gave place to batters with low apparent viscosity values. During heating, bubbles underwent a marked expansion in replaced cakes if compared to the control cake. The low batter stability in fat-replaced samples increased bubble movement, giving place to cakes with bigger cells and less height than the control. Sugar-replaced samples had smaller and fewer cells and lower height than the control. Moreover, sugar replacement decreased hardness and cohesiveness and in- creased springiness, which could be related with a denser crumb and an easily crumbled product. Regarding the sensory analysis, a replacement up to 50 % of fat and 30 % of sugar, separately and simultaneously, did not change remarkably the overall acceptability of the cakes. However, the sponginess and the sweetness could be improved in all the replaced cakes, according to the Just About Right scales.

Functionality of lipase and emulsifiers in low-fat cakes with inulin

The functional effects of lipase (0.003 and 0.006 g/100 g of flour) and emulsifier (0.5 and 1 g/100 g of flour) on fat-replaced (0%, 50% and 70%) batters and cakes with inulin (0, 7.5 and 10 g/100 g/of flour, respectively) were studied. Emulsifier addition significantly lowered the relative density of the batter. Emulsifier incorporation increased the viscoelastic properties of the batter. In contrast, lipase incorporation decreased the degree of system structuring. The evolution of the dynamic moduli and complex viscosity with rising temperatures were studied. Batters with 1 g/100 g emulsifier displayed a significantly lower complex viscosity during heating, resulting in collapsed cakes. Differential scanning calorimetry results revealed that the thermal setting in the control cakes occurred at higher temperatures, and accordingly, greater cake expansion was observed. Cakes with 0.003 g/100 g lipase or 0.5 g/100 g emulsifier displayed volume and crumb cell structure that were similar to those of control cakes. Higher concentrations of both improvers gave rise to cakes with lower volume, higher hardness and lower springiness. During storage time, cakes with lipase displayed lower hardness. Both improvers, at low concentrations, could improve certain physical characteristics, such as crumb structure, of fat-replaced cakes with inulin.

Changes in sewage sludge carbon forms along a treatment stream

The behaviour and fate of macronutrients and pollutants in sewage sludge applied to the land are affected by the chemical composition of the sludge organic matter, which in turn is influenced by both sewage source and by sewage treatment processes. In this study, 13C nuclear magnetic resonance (NMR) spectroscopy was used to characterise the organic matter of sludges collected at three different points along the treatment stream of a municipal sewage works with a domestic catchment. Sludge at the first point, an undigested liquid (UL) sludge, had a substantially different composition to the anaerobically digested (AD) and dewatered sludge cake (DC) materials, which were similar to each other. In particular, the UL sludge contained more alkyl C than the AD or DC sludges. All three sludges were found to contain mobile alkyl C that is poorly observed using the cross polarisation (CP) technique, necessitating the use of the less sensitive, but more quantitatively reliable direct polarisation (DP) technique to obtain accurate distributions of C types.

Some potential inaccuracies of the p -nitrophenyl phosphomonoesterase assay in the study of the phosphorus nutrition of soil borne fungi

The p-nitrophenol phosphomonoesterase assay (pNPPase) is commonly used to measure cell-wall-associated and extracellular phosphatase activity of soil fungi. pNPPases are usually assayed in the context of fungal nutrition, where inorganic P supply might be enhanced by the mineralisation of organic P sources in the soil. We report here on a series of experiments with the ectomycorrhizal basidiomycete Hebeloma cylindrosporum that highlight components of accepted methodology that might impinge on the reliability of the assay. These include the loss of pNPPase after filtration, inaccuracies in measuring wall-associated enzyme and the ample pool of intracellular pNPPase can be mistakenly measured as external pNPPase if cells are accidentally damaged.

Nitrogen dynamics under Lolium perenne after a single application of different sewage sludge types from the same treatment stream

Three sludge types from the same treatment stream (undigested liquid, anaerobically digested liquid and dewatered, anaerobically digested cake) were used in a field based tub study. Amendments (4, 8, and 16 Mg dry solid (ds)ha(-1)) were incorporated into the upper 15 cm of a sandy loam soil prior to sowing with rye-grass (Lolium perenne L.). Nitrogen transformations in the soil were determined for the 80 d period following incorporation. Nitrogen uptake and crop yield were measured in the cut sward 35 and 70 d after sowing. The study showed that application of sewage sludge at rates as low as 4 Mgha(-1) can have a nutritional benefit to rye-grass over the two harvests. Differences in N transformation, and hence crop nutritional benefit, between sludge types were evident throughout the experiment. In particular, the dewatering process changed the mineral N characteristics of the anaerobically digested sludge, which, when not dewatered, outperformed the other sludges in terms of yield and mineralisation rate at both harvests. The dewatered sludge produced the lowest yield of rye-grass. The undigested liquid sludge had the lowest foliar N and soil NO(3)-N concentrations, possibly immobilised as the large oxidisable C component of this sludge was metabolised by the microbial biomass. Correlation data support the concept of preferential uptake of NH(4)-N over NO(3)-N in Lolium perenne. Results are discussed in the context of managing sludge type and application for a plant nutrient source and NO(3)-N release.

Astringency reduction in red wine by whey proteins

Whey is a by-product of cheese manufacturing and therefore investigating new applications of whey proteins will contribute towards the valorisation of whey and hence waste reduction. This study shows for the first time a detailed comparison of the effectiveness of gelatin and β-lactoglobulin (β-LG) as fining agents. Gelatin was more reactive than whey proteins to tannic acid as shown by both the astringency method (with ovalbumin as a precipitant) and the tannins determination method (with methylcellulose as a precipitant). The two proteins showed similar selectivity for polyphenols but β-LG did not remove as much catechin. The fining agent was removed completely or to a trace level after centrifugation followed by filtration which minimises its potential allergenicity. In addition, improved understanding of protein–tannin interactions was obtained by fluorescence, size measurement and isothermal titration calorimetry (ITC). Overall this study demonstrates that whey proteins have the potential of reducing astringency in red wine and can find a place in enology.