Effect of transglutaminase-catalyzed polymerization of beta-casein on its emulsifying properties

beta-Casein was polymerized to various extents by the transglutaminase-catalyzed cross-linking reaction. Under the reaction conditions used, dimer, trimer, and tetramer of beta-casein were produced as predominant species at short reaction times (20-90 min) and large polymers at longer reaction times (3-24 h). Examination of the emulsifying properties of these cross-linked beta-casein polymers showed that although the emulsifying activity index decreased, the storage stability of the emulsions increased with increasing degree of polymerization. Experiments with mixtures of native and polymerized beta-casein also showed that increasing the fraction of polymerized beta-casein in the mixture increased the emulsion stability. This enhancement of emulsion stability might be due either to an enhancement in steric stabilization, attributable to the branched nature of the trasgutaminase-catalyzed polymers, or to stronger cohesive interactions between polymerized beta-casein molecules than between monomeric beta-casein molecules in the film.     

Protein Composition‐Functionality Relationships for a Set of Argentinean Wheats

Relationships between flour functional properties and protein composition were studied using a set of 138 Argentinean wheat samples. Among different protein groups, the incremental increase of gliadin with increasing grain protein content was highest followed by polymeric protein with albumin‐globulin content much lower. Functional properties could be divided into two groups based on dependence on protein composition. Properties such as dough extensibility and bake test loaf volume correlated highly with the percentage of polymeric protein in the grain. Properties such as mixograph dough development time were best correlated with the percentage of polymeric protein in the protein (PPP). Alveograph tenacity showed no significant dependence on PPP. as found previously for extensigraph maximum resistance, but it was correlated with the percentage of unextractable polymeric protein in the protein. Energy (W) appeared to be a more useful alveograph parameter for predicting flour quality.     

Relationships between dairy powder surface composition and wetting properties during storage: importance of residual lipids

The relationships between powder surface composition and powder rehydration properties under variable conditions of storage are investigated in this paper. A rheological approach was used to evaluate the modifications induced by storage on the rehydration properties of native phosphocaseinate powder. Concurrently, the powder surface composition (i.e., lactose, proteins, and lipids) was evaluated by X-ray photoelectron spectroscopy (XPS). A strong correlation was found between the powder wetting time lengthening and the migration of lipids on the powder surface during storage. XPS studies indicated also an over-representation of lipids on the powder surface (6%) in comparison with total lipids (0.4%) even on fresh powder before storage. Detailed investigation of powder lipids revealed the presence of high levels of polar lipids (66% compared with <1% in milk lipids). Their amphiphilic nature and their melting points could explain the extensive enrichment of lipids observed at the powder surface during processing and storage.     

The spatial heterogeneity of chemical properties in acid forest soils and its importance for tree nutrition

For two sites in the Black Forest, presence, extent and ecological significance of structure-dependent chemical. heterogeneity in the rhizosphere of acid forest soils is discussed. The heterogeneity is demonstrated by two independent methods. First, by comparison of soil solution composition in (quasi-) equilibrium with undisturbed soil cores and sieved soil samples. Water soluble K, Ca and Mg show a selective depletion on aggregate surfaces. Second, by comparison of cumulated amounts of exchangeable ions obtained from percolated homogenized soil samples and undisturbed soil cores. The release of exchangeable K, Ca and Mg is less, if soil structure is not destroyed. The finding that aggregate surfaces are more depleted of exchangeable and water-soluble K, Ca and Mg is common in aggregated acid forest soils. As aggregate surfaces bind soil water with low suction, that is where the roots preferentially grow. Consequently, a considerable underestimation of chemical stress in the rhizosphere is possible, if soil structure is destroyed for analysis.     

Dynamics of competitive adsorption of alphas-casein and beta-casein at planar triolein-water interface: evidence for incompatibility of mixing in the interfacial film

Competitive adsorption of alpha(s)-casein and beta-casein from a bulk solution mixture to the triolein-water interface has been studied. Although the binding affinity of alpha(s)-casein to the triolein-water interface was lower than that of beta-casein in single-component systems, in a 1:1 mixture of alpha(s)-casein and beta-casein in the bulk solution the ratio of interfacial concentrations of alpha(s)-casein to beta-casein at equilibrium was about 2:1, indicating that alpha(s)-casein was preferentially adsorbed to the triolein-water interface. Furthermore, the equilibrium composition of alpha(s)-casein and beta-casein in the interfacial film at various bulk concentration ratios did not follow a simple Langmuir adsorption model. This deviation from ideal behavior was mainly due to thermodynamic incompatibility of mixing of these caseins in the interfacial region. The value of the incompatibility parameter, X(12), for these caseins at the triolein-water interface was much greater than that at the air-water interface. Displacement experiments showed that while alpha(s)-casein could dynamically displace beta-casein when the latter was in an unsaturated monolayer state at the interface, it could not do so when beta-casein was in a saturated monolayer film state. It is hypothesized that, because of thermodynamic incompatibility of mixing, the alpha(s)-casein and beta-casein mixed film at the oil-water interface may undergo two-dimensional phase separation.     

Proteolytic activities of suparen and rennilase on buffalo, cow, and goat whole casein and beta-casein.

The proteolytic specificity and activity of Mucor miehei protease (Rennilase) and Endothia parasitica protease (Suparen) on buffalo, cow, and goat whole casein and beta-casein (CN) were studied by analyzing the degradation products. The results suggest that Rennilase hydrolyzes casein of the three species in a manner similar to that of chymosin, resulting in the formation of alpha(s1)-I and beta-I, -II, -III as initial degradation fragments of alpha(s1)- and beta-CN. alpha(s1)-I was also the initial breakdown product of alpha(s1)-CN by Suparen. Contrary to Rennilase, Suparen showed a higher affinity toward beta-CN and hydrolyzes beta-CN, giving rise to degradation products characterized by mobility lower than that of beta-CN. Increasing NaCl concentration (>3%) reduced the proteolysis of beta-CN of the three species by Rennilase but not by Suparen. The hydrolysis of alpha(s1)-CN and alpha(s1)-I by the two enzymes was enhanced in the presence of NaCl.     

Physical properties of homoionic montmorillonite and kaolinite complexed with amino acids and peptides

X-ray diffractometry was used to investigate whether various amino acids and peptides expanded montmorillonite (M) homoionic to different cations, and the electrophoretic mobility (EPM) was measured to determine whether the binding of these amino acids and peptides influenced the net surface charge of M and kaolinite (K). Neither aspartic acid, adsorbed at equilibrium at concentrations of 20–250 nM mg^?1 M homoionic to Ca or Zn, nor cysteine, at 50–830 nM mg^?1 M homoionic to Zn or Al, caused expansion, whereas proline, at 90–870 nM mg^?1 clay, caused expansion of M homoionic to Ca or Zn, and both proline and arginine, at 60–1150 nM mg^?1 clay, caused expansion of M homoionic to H or Al. The extent of expansion caused by proline and arginine was related to the concentration of the amino acids and the exchangeable cation on the homoionic M. Only M-A1 complexed with arginine remained expanded after extensive washing of the clay-amino acid complexes. The expansion of M was apparently dependent on both the type of exchangeable cation on the clay and the characteristics of the amino acid (i.e. the functional groups, dielectric constant increment, dipole moment and concentration). M homoionic to H, Na, Zn, Al or La was not expanded by a series of di- to tetrapeptides bound on these clays and none of the homoionic K-amino acid or peptide complexes was expanded. The EPM of the homoionic M and K and of the corresponding clay-amino acid or peptide complexes was identical, indicating that the concentration of the amino acids or peptides bound was not sufficient to cover enough of the surface of the clays to alter significantly their net charge. The lack of differences in the bulk pH of the clays after binding with amino acids or peptides (with the exception of complexes with arginine) also indicated limited coverage of the clays by these organics.     

Analytical Techniques for Understanding Nixtamalized Corn Flour: Particle Size and Functionality Relationships in a Masa Flour Sample

Instant masa flour finds extensive use in the food industry for making tortillas, taco shells, tamales, corn chips, and tortilla chips, and as an ingredient in extruded snacks. Due to lack of standard techniques for measuring masa functionality, processors and end‐users use masa flour particle‐size distribution and rheological characteristics in an attempt to predict its end use. In this study, a commercial masa flour sample was characterized by fractionating on the basis of particle size. Physicochemical and functional properties of masa flour fractions were investigated to establish structure‐composition and functionality relationships. It was observed that Rapid Visco Analyser (RVA) pasting profiles of flour fractions and textural properties of dough prepared on rehydration were related to particle size, yet, upon regrinding, RVA profiles did not change as markedly as expected. Differences in RVA measurements of the sized fractions could not be explained on the basis of hydration rate or total starch content. It was concluded that masa dough textural and RVA characteristics may be influenced by the status of starch polymer structures formed during nixtamalization.     

Production of porcine hemoglobin peptides at moderate temperature and medium pressure under a nitrogen stream. Functional and antioxidant properties

A new hydrolysis method for producing peptides from porcine hemoglobin has been developed. Current processes are based on the use of expensive enzymes or high hydrostatic pressures. In the present study, a cheap and effective process has been assayed to produce peptides from purified porcine hemoglobin. A solution of purified hemoglobin is heated at different temperatures and pressurized at 4 MPa while a stream of nitrogen is injected into the reactor. A total of 82% of initial hemoglobin was transformed into peptides presenting an average size of 3.2 kDa. Some preferential hydrolyzed bonds have been detected. The peptide size distribution was evaluated at different times and temperatures. It has been demonstrated that this technique produces large amounts of peptides possessing good antioxidant properties. Furthermore, functional properties are conserved, and a desirable decrease in color (80%) is achieved.     

Quantitative analysis of pyroglutamic acid in peptides.

A simplified and rapid procedure for the determination of pyroglutamic acid in peptides was developed. The method involves the enzymatic cleavage of an N-terminal pyroglutamate residue using a thermostable pyroglutamate aminopeptidase and isocratic HPLC separation of the resulting enzymatic hydrolysate using a column switching technique. Pyroglutamate aminopeptidase from a thermophilic archaebacteria, Pyrococcus furiosus, cleaves N-terminal pyroglutamic acid residue independent of the molecular weight of the substrate. It cleaves more than 85% of pyroglutamate from peptides whose molecular weight ranges from 362.4 to 4599.4 Da. Thus, a new method is presented that quantitatively estimates N-terminal pyroglutamic acid residue in peptides.     

Effect of simulated gastrointestinal digestion on the antihypertensive properties of ACE-inhibitory peptides derived from ovalbumin

Food-derived bioactive peptides with ACE-inhibitory properties are receiving special attention due to their beneficial effects in the treatment of hypertension. In this work we evaluate the impact of a simulated gastrointestinal digestion on the stability and activity of two bioactive peptides that derive from ovalbumin by enzymatic hydrolysis, YAEERYPIL and RADHPFL. These peptides possess in vitro ACE-inhibitory activity and antihypertensive activity in spontaneously hypertensive rats (SHR). The results showed that YAEERYPIL and RADHPFL were susceptible to proteolytic degradation after incubation with pepsin and a pancreatic extract. In addition, their ACE-inhibitory activity in vitro decreased after the simulated digestion. The antihypertensive activity on SHR of the end products of the gastrointestinal hydrolysis, YAEER, YPI, and RADHP, was evaluated. The fragments YPI and RADHP significantly decreased blood pressure, 2 h after administration, at doses of 2 mg/kg, but they probably did not exert their antihypertensive effect through an ACE-inhibitory mechanism. It is likely that RADHP is also the active end product of the gastrointestinal digestion of the antihypertensive peptides FRADHPFL (ovokinin) and RADHPF (ovokinin 2-7).     

Iron tissue storage and hemoglobin levels of deficient rats repleted with iron bound to the caseinophosphopeptide 1-25 of beta-casein.

Caseinophosphopeptides (CPP) issued from enzyme digestion of caseins bind cations and keep them soluble in the digestive tract. They could be used as ligands to improve iron (Fe) bioavailability. Fe-deficient young rats were repleted with Fe (40 or 200 mg/kg of diet) bound either to the beta-CN (1-25) of beta-casein or to whole beta-casein or as FeSO(4). A control pair-fed group was given 200 mg of Fe (FeSO(4))/kg of diet for 6 weeks. After repletion, hemoglobin concentration of the control group was reached only by the ) animals fed 200 mg of Fe/kg; beta-CN (1-25) bound Fe (40 and 200 mg) produced higher Fe liver and spleen stores than FeSO(4). Binding Fe to the whole, nonhydrolyzed beta-casein gave results intermediate between the other experimental groups. Binding Fe to phosphoserine residues of low molecular weight CPP improved its ability to cure anemia and to restore iron tissue stores, as compared to Fe bound to the whole casein and to inorganic salts.     

Antioxidant properties of casein calcium peptides and their effects on lipid oxidation in beef homogenates

The antioxidant activity of casein calcium peptides in several in vitro assay systems was investigated. Casein calcium peptides were prepared by the microbial enzymic hydrolysis of casein calcium. The main peak of the molecular mass distribution of the peptides was about 3 kDa. Casein calcium peptides showed strong antioxidant activity with the beta-carotene bleaching method, and they also showed scavenging activity against radicals such as superoxide radicals, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, and hydroxyl radicals. Antioxidant activity was increased with an increasing peptide concentration. Casein calcium peptides also showed strong antioxidant activity against lipid oxidation in ground beef homogenates. These results suggest that casein calcium peptides are a suitable natural antioxidant that prevents the lipid oxidation of meat and related food ingredients.     

Characterization of peptides formed during fermentation of cocoa bean.

Analysis by SDS-PAGE and GPC-MS of fermented cocoa extracts shows changes in the amount and composition of the major proteins, accompanied by formation of complex distributions of peptides. MS/MS studies and application of SEQUEST sequencing software have allowed identification of two related peptides, a hexapeptide and a nonapeptide, formed from vicilin, one of the cocoa storage proteins. Time course studies of the two peptides show different abundance profiles and indicate, in part, production of the hexapeptide from the nonapeptide.     

Use of palm-mat geotextiles for soil conservation: I. Effects on soil properties

Despite geotextile-mats having the potential for soil conservation, field studies on the effects of geotextiles on soil properties are limited. Hence, the utilization of palm-mat geotextiles as a potential soil conservation technique was investigated at Hilton, east Shropshire, U.K. (52°33′5.7″ N, 2°19′18.3″ W). Geotextile-mats constructed from Borassus aethiopum (Borassus palm of West Africa) and Mauritia flexuosa (Buriti palm of South America) leaves are termed Borassus mats and Buriti mats, respectively. Field experiments were conducted at Hilton during 2007–2009, to study the impacts of Borassus and Buriti mats on selected properties of the topsoil (0–5 cm). Ten fixed plots (10 × 1 m on a 15° slope) were established, with duplicate treatments. The treatments were: (i) bare soil; (ii) permanent grassed; (iii) bare soil with 1 m Borassus-mat buffer strips (area coverage ~ 10%) at the lower end of the plots; (iv) bare soil with 1 m Buriti mat buffer strips (area coverage ~ 10%) at the lower end of the plots; and (v) completely-covered with Borassus mats. Initial and final soil samples of the topsoil were collected and analysed for bulk density, aggregate stability, soil organic matter (SOM), total soil C (TSC), total soil N (TSN) and pH. Results indicate that, apart from Borassus completely-covered plots, soil bulk density increased and aggregate stability decreased in all plots after two years. Despite decreases in SOM contents in bare plots, SOM content did not change after two years in the grassed and geotextile treated plots. Treatments had no effects on changes in pH, TSC or TSN. Both Borassus and Buriti mat-covers within the buffer strip plots had little impact on SOM, TSC and TSN changes compared with bare soils within the same plots. Thus, Borassus buffer strip plots were very effective in maintaining some soil properties (i.e. SOM, TSC, and TSN) after two years of erosion by water. In summary, utilization of Borassus mats as buffer strips was very successful in conserving soil properties on a loamy sand soil.     

Use of Two Endoxylanases with Different Substrate Selectivity for Understanding Arabinoxylan Functionality in Wheat Flour Breadmaking

A Bacillus subtilis endoxylanase (XBS) with a strong selectivity for hydrolysis of water‐unextractable arabinoxylan (WU‐AX) and an Aspergillus aculeatus endoxylanase (XAA) with a strong selectivity for hydrolysis of water‐extractable arabinoxylan (WE‐AX) were used in straight‐dough breadmaking with two European wheat flours. Dough, fermented dough, and bread characteristics with different levels of enzyme addition were evaluated with a strong emphasis on the arabinoxylan (AX) population. The WU‐AX solubilized by XBS during breadmaking were mainly released during mixing and had higher molecular weight, in contrast to their counterparts solubilized by XAA, which were mainly released during fermentation and had lower molecular weight. This coincided with increased loaf volume with XBS and a negative to positive loaf volume response with XAA. Bread firmness and dough extract viscosity also were affected by endoxylanase addition. Results confirmed that WU‐AX are detrimental for breadmaking, while WE‐AX and solubilized AX with medium to high molecular weight have a positive impact on loaf volume.     

Fractionation of beta-lactoglobulin tryptic peptides by ampholyte-free isoelectric focusing.

Solutions of tryptic hydrolysate of bovine beta-lactoglobulin were fractionated by liquid-phase IEF in a preparative Rotofor cell at constant power for 2 h without ampholytes in order to identify interactions between peptides. The 20 peptide fractions collected were analyzed by capillary electrophoresis and SDS-PAGE under native, denaturing, and reducing conditions. The hydrolysate was shown to be composed mainly of acidic peptides (pI 2-5, 62%) of molecular mass below 6 kDa, and numerous disulfide bonds were detected. Purified peptides (beta-LG 15-20, 71-75, 76-82, and 84-91) were also focused individually and in mixtures and matched to components of the IEF fractions obtained from the tryptic hydrolysate of beta-LG. The separation of acidic (beta-LG 84-91) and basic (beta-LG 76-82) peptides was achieved by IEF, whereas uncharged peptides (beta-LG 15-20 and 71-75) were poorly separated due to their low electrophoretic mobility. Because no peptide-peptide interaction could be identified by IEF fractionation, it is suggested that electrical fields may decrease electrostatic interactions between charged peptides.     

Spatial variability of Southeastern U.S. Coastal Plain soil physical properties: Implications for site-specific management

Our objectives were to describe the field-scale horizontal and vertical spatial variability of soil physical properties and their relations to soil map units in typical southeastern USA coastal plain soils, and to identify the soil properties, or clusters of properties, that defined most of the variability within the field. The study was conducted on a 12-ha field in Kinston, NC. A 1:2400 scale soil survey had delineated three soil map units in the field: Norfolk loamy sand, Goldsboro loamy sand, and Lynchburg sandy loam. These are representative of millions of hectares of farmland in the Coastal Plain of the southeastern USA. Sixty soil cores were taken to ∼ 1-m depth, sectioned into five depth increments, and analyzed for: soil texture as percentage sand, silt, and clay; soil water content (SWC) at − 33 and − 1500 kPa; plant available water (PAW); saturated hydraulic conductivity (Ksat); bulk density (BD); and total porosity. A penetrometer was used to measure cone index (CI) at each sample location. Variography, two mixed-model analyses, and principal components analysis were conducted. Results indicated that soil physical properties could be divided into two categories. The first category described the majority of the within-field variability and included particle size distribution (soil texture), SWC, PAW, and CI. These characteristics showed horizontal spatial structure that was captured by soil map units and especially by the division between sandy loams and finer loam soils. The second class of variables included BD, total porosity, and Ksat. These properties were not spatially correlated in the field and were unrelated to soil map unit. These findings support the hypothesis that coastal plain soil map units that delineate boundaries between sandy loams versus finer loam soils may be useful for developing management zones for site-specific crop management.     

Transboundary conservation in the greater Virunga landscape: Its importance for landscape species

Several of the protected areas within the Albertine Rift are contiguous with protected areas across international boundaries. This is particularly true for the Greater Virunga Landscape, which includes Virunga National Park in the Democratic Republic of Congo (DRC) and ten contiguous protected areas in Uganda and Rwanda. The larger bodied animal species that occur in this landscape have moved freely across what are now international borders for millennia and some species probably need this larger landscape if their populations are to remain viable. An analysis was carried out to identify these ‘landscape species’ and the importance of this cross-border movement is assessed in the light of civil wars in the region. For 13 years the International Gorilla Conservation Programme has been working in the Virunga Volcanoes and Bwindi Impenetrable National Park to foster transboundary collaboration. The results have shown that regular meetings and planning of activities by wardens can lead to better conservation even with countries at war with each other. More recently the Wildlife Conservation Society has started a programme to support transboundary collaboration further north in the landscape so that all contiguous protected areas are working together. The results show that mountain gorilla numbers have on the whole increased during the past 25 years despite civil wars in the region and this can largely be attributed to their ability to generate income from tourism but also to enhanced transboundary collaboration between Congo, Rwanda and Uganda. Ungulate numbers on the other hand have declined drastically since the 1960s but it is shown that the connectivity in the landscape has been important in reducing the impact of the civil war on elephants.