Gelation of chicken pectoralis major myosin and heat-denatured beta-lactoglobulin

Thermal, rheological, and microstructural properties of myosin (1 and 2% protein) were compared to mixtures of 1% myosin and 1% heat-denatured beta-lactoglobulin aggregates (myosin/HDLG) and 1% myosin and 1% native beta-lactoglobulin (myosin/beta-LG) in 0.6 M NaCl and 0.05 M sodium phosphate buffer, pH 6.0, 6.5, and 7.0 during heating to 71 degrees C. Thermal denaturation patterns of myosin and myosin/HDLG were similar except for the appearance of an endothermic peak at 54-56 degrees C in the mixed system. At pH 7.0, 2% myosin began to gel at 48 degrees C and had a storage modulus (G') of 500 Pa upon cooling. Myosin/HDLG (2% total protein) had a gel point of 48 degrees C and a G' of 650 Pa, whereas myosin/beta-LG had a gel point of 49 degrees C but the G' was lower (180 Pa). As the pH was decreased, the gel points of myosin and myosin/HDLG decreased and the G' after cooling increased. The HDLG was incorporated within the myosin gel network, whereas beta-LG remained soluble.     

Light-scattering study of the structure of aggregates and gels formed by heat-denatured whey protein isolate and beta-lactoglobulin at neutral pH

The structure of aggregates and gels formed by heat-denatured whey protein isolate (WPI) has been studied at pH 7 and different ionic strengths using light scattering and turbidimetry. The results were compared with those obtained for pure beta-lactoglobulin (beta-Lg). WPI aggregates were found to have the same self-similar structure as pure beta-Lg aggregates. WPI formed gels above a critical concentration that varied from close to 100 g/L in the absence of added salt to about 10 g/L at 0.2 M NaCl. At low ionic strength (<0.05 M NaCl) homogeneous transparent gels were formed, while at higher ionic strength the gels became turbid but had the same self-similar structure as reported earlier for pure beta-Lg. The length scale characterizing the heterogeneity of the gels increased exponentially with increasing NaCl concentration for both WPI and pure beta-Lg, but the increase was steeper for the former.     

Enterocyte and M-cell transport of native and heat-denatured bovine beta-lactoglobulin: significance of heat denaturation

The three-dimensional structure, digestibility, and immunological properties of bovine beta-lactoglobulin (beta-lg) are modified by heat treatments used in processing of liquid milk products. Because it is not known if such treatments also modify the intestinal transport properties of beta-lg, the transport of native and heat-denatured bovine beta-lg was investigated in experimental cell models using Caco-2 cells and M cells. Transport of beta-lg labeled with a fluorescent marker was followed with fluorometric measurements, electrophoretic analyses, and fluorescence microscopy. The data show that both cell types transported native beta-lg more efficiently than they did heat-denatured beta-lg. In addition, M cells transported native beta-lg more than Caco-2 cells. Transport of native and heat-denatured beta-lg was transcellular. The electrophoretic data also suggest that heat-denatured beta-lg may have degraded more than native beta-lg during the transport.     

Riboflavin-sensitized photochemical changes in beta-lactoglobulin in an aqueous buffer solution as affected by ascorbic acid

The effects of ascorbic acid on the riboflavin-sensitized photochemical changes in beta-lactoglobulin in an aqueous buffer solution as determined by high performance gel permeation liquid chromatography (HPGPLC), insoluble protein content, and individual amino acid content during fluorescent light illumination were studied. The riboflavin-sensitized photochemical degradation of beta-lactoglobulin was effectively inhibited by ascorbic acid, and its inhibitory effectiveness was concentration dependent. The 0.1% ascorbic acid treatment showed 74.4% inhibition of beta-lactoglobulin degradation as determined by a HPGPLC during 6 h light illumination. Insolubility of beta-lactoglobulin in a buffer solution during light illumination was also effectively decreased by ascorbic acid treatment. The riboflavin-sensitized photochemical reduction of cysteine, histidine, lysine, methionine, and tryptophan in beta-lactoglobulin was high during 6 h fluorescent light illumination. The 0.1% ascorbic acid treatment exhibited 20.8% inhibition of total amino acid degradation in beta-lactoglobulin during 6 h light illumination, showing strong inhibitory activity against the degradation of arginine, aspartic acid, cystein, glycine, histidine, phenylalanine, proline, serine, and tryptophan.     

Identification of peptides in aggregates formed during hydrolysis of beta-lactoglobulin B with a Glu and Asp specific microbial protease

The purpose of the present study was to identify the peptides responsible for aggregate formation during hydrolysis of beta-lactoglobulin by BLP at neutral pH. Hydrolysates taken at various stages of aggregate formation were separated into a precipitate and a soluble phase and each was analyzed by CE and mass spectrometry. The aggregates consisted of six to seven major peptides of which four were tentatively identified. The peptides were positively charged at neutral pH and had a high charge-to-mass ratio at low pH. The fragment f135-158 seemed to be the initiator of aggregation, since it was present at high concentration in the aggregates at all stages, and the concentration of this peptide remained low in the supernatant. F135-158 contains several basic and acid amino acids alternating with hydrophobic amino acids, which is in accordance with formation of noncovalently linked aggregates, as previously shown.     

Heat-induced aggregation of beta-lactoglobulin as a function of pH.

The effect of pH in the range 6.0-8.0 on the denaturation and aggregation of beta-lactoglobulin (beta-lg) was investigated. Results were interpreted in terms of the reaction scheme for the denaturation and aggregation of beta-lg proposed by Roefs and De Kruif (Eur. J. Biochem. 1994, 226, 883-889). The rate of conversion of native beta-lg increased strongly at higher pH values, whereas the molecular mass of the aggregates decreased strongly. In the pH range 6.4-8.0 aggregates were formed mainly by intermolecular disulfide bonds, but even at pH 6.0, thiol/disulfide exchange reactions were involved, although to a lesser extent. The time course of the exposure of the thiol group in native beta-lg upon heating and the subsequent disappearance of this group through the formation of disulfide-linked aggregates was investigated by reaction with 5,5'-dithiobis(2-nitrobenzoic acid) and varied strongly with pH. These observations could be used, in combination with the reaction steps of the reaction scheme, to describe qualitatively the strongly pH-dependent isothermal calorimetry curves, measured at 65 degrees C.     

Denaturation of beta-lactoglobulin in pressure-treated skim milk

The kinetics of beta-lactoglobulin (beta-LG) denaturation in pressure-treated reconstituted skim milk samples over a wide pressurization range (100-600 MPa) and at various temperatures (10-40 degrees C) was studied. Denaturation was extremely dependent on the pressure and duration of treatment. At 100 MPa, no denaturation was observed regardless of the temperature or the holding time. At higher pressures, the level of denaturation increased with an increasing holding time at a constant pressure or with increasing pressure at a constant holding time. At 200 MPa, there was only a small effect of changing the temperature at pressurization. However, at higher pressures, increasing the temperature from 10 to 40 degrees C markedly increased the rate of denaturation. The two major genetic variants of beta-LG (A and B) behaved similarly to pressure treatment, although the B variant appeared to denature slightly faster than the A variant at low pressures (< or =400 MPa). The denaturation could be described as a second-order process for both beta-LG variants. There was a marked change in pressure dependence at about 300 MPa, which resulted in markedly different activation volumes in the two pressure ranges. Evaluation of the kinetic and thermodynamic parameters suggested that there may have been a transition from an aggregation-limited reaction to an unfolding-limited reaction as the pressure was increased.     

Semi-mechanistic partial buffer approach to modeling pH, the buffer properties, and the distribution of ionic species in complex solutions

In many biological science and food processing applications, it is very important to control or modify pH. However, the complex, unknown composition of biological media and foods often limits the utility of purely theoretical approaches to modeling pH and calculating the distributions of ionizable species. This paper provides general formulas and efficient algorithms for predicting the pH, titration, ionic species concentrations, buffer capacity, and ionic strength of buffer solutions containing both defined and undefined components. A flexible, semi-mechanistic, partial buffering (SMPB) approach is presented that uses local polynomial regression to model the buffering influence of complex or undefined components in a solution, while identified components of known concentration are modeled using expressions based on extensions of the standard acid-base theory. The SMPB method is implemented in a freeware package, (pH)Tools, for use with Matlab. We validated the predictive accuracy of these methods by using strong acid titrations of cucumber slurries to predict the amount of a weak acid required to adjust pH to selected target values.     

Glycation and phosphorylation of beta-lactoglobulin by dry-heating: effect on protein structure and some properties

Beta-lactoglobulin (beta-Lg) was glycated with maltopentaose and subsequently phosphorylated by dry-heating in the presence of pyrophosphate to investigate the structural and functional properties of phosphorylated beta-Lg. The circular dichroism spectra showed that the change of the secondary structure in the beta-Lg molecule by glycation and subsequent phosphorylation was small. The differential scanning calorimetry thermograms of beta-Lg showed that the denaturation temperature of the most stable domain was only slightly affected, whereas the retinol-binding activity of beta-Lg was somewhat reduced by glycation and subsequent phosphorylation. These results indicated that the conformational changes of the beta-Lg molecule by glycation and subsequent phosphorylation were mild. The anti-beta-Lg antibody response was somewhat reduced by glycation, but significant changes were not observed by phosphorylation. Although the stability of beta-Lg against heat-induced insolubility was improved by glycation alone, it was further enhanced by phosphorylation. The calcium phosphate solubilizing ability of beta-Lg was enhanced by phosphorylation following glycation.     

Effects of high-pressure processing at low temperature on the molecular structure and surface properties of beta-lactoglobulin

High-pressure processing (HPP) was utilized to induce unfolding of beta-lactoglobulin (beta-LG). beta-Lactoglobulin solutions at concentrations of 0.5 mg/mL, in pH 7.5 phosphate buffer, were pressure treated at 510 MPa for 10 min at either 8 or 24 degrees C. The secondary structure, as determined by circular dichroism (CD), of beta-LG processed at 8 degrees C appeared to be unchanged, whereas beta-LG processed at 24 degrees C lost alpha-helix structure. Tertiary structures for beta-LG, as determined by near-UV CD, intrinsic protein fluorescence spectroscopy, hydrophobic fluorescent probe binding, and thiol group reactivity, were changed following processing at either temperature. The largest changes to tertiary structure were observed for the samples processed at 24 degrees C. Model solutions containing the pressure-treated beta-LG showed significant decreases in surface tension at liquid-air interfaces with values of 54.00 and 51.69 mN/m for the samples treated at 24 and 8 degrees C, respectively. In comparison, the surface tension for model solutions containing the untreated control was 60.60 mN/m. Changes in protein structure during frozen and freeze-dried storage were also monitored, and some renaturation was observed for both storage conditions. Significantly, the sample pressure-treated at 8 degrees C continued to display the lowest surface tension.     

Earthworm effects on selected physical and chemical properties of soil aggregates

Summary Some physical and chemical properties of 1-to 2-mm aggregates obtained from casts and the burrow-wall material of the earthworm species Lumbricus terrestris, Aporrectodea longa, and Aporrectodea caliginosa were determined in order to show the effects of earthworms on the stabilization of soil aggregates. The results were compared with those of the natural soil from the Ap horizon of a Parabraunerde (Luvisol, FAO). Both the tensile strength and the water stability of aggregates from casts and burrow-wall material were reduced compared with those of the natural aggregates but were increased compared with those of remoulded aggregates. These results indicate that to a great extent existing bonds are destroyed by earthworm ingestion. Nevertheless, earthworm activities are advantageous for the stabilization of reformed aggregates. The coarse sand fraction is reduced by selective ingestion by earthworms. The organic C content is increased by 4.1–21.0% for burrow-wall material and by 21.2–43.0% for casts. The carbonate content of aggregates from casts and burrow-wall material of L. terrestris was reduced by more than 50%, while that of A. longa showed no noticeable changes and that of A. caliginosa was increased by more than 60%. The total content of polysaccharides was increased by 35–87% for casts and by 33–46% for the burrow-wall material of all earthworm species. The most frequently detected monosaccharides were glucose, galactose, and glucosamine. L. terrestris appeared to have the strongest effect on the interparticle bonding of the reformed aggregates, measured both as tensile strength and water stability, followed by A. longa and A. caliginosa.     

Reclamation of a fine clayey montmorillonitic soil with high-salt water under laboratory conditions

An impermeable fine clayey montmorillonitic soil (Aquic Xerofluvent) was reclaimed using high-salt water and the addition of a constant quantity of Ca at each step in successive dilutions. The soil attained the theoretically expected values of ESP and the depth of water which passed through the soil was close to the theoretical level. The leachate was studied throughout the process of reclamation and the following results obtained: At the end of the reclamation all ions were seen to be decreasing. The percolation rate of the soil and the ?valence-dilution”? effect of the reclaiming water permitted a satisfactory exchange of Na by Ca and Mg keeping the quantity of water close to the theoretically expected levels. The mean hydraulic conductivity (MHC) is strongly depended on the electrolyte concentration and the sodium/total concentration (Na/C) or Ca+Mg/total concentration (R) values of the leachate.     

Dielectric study of heat-denatured ovalbumin in aqueous solution by time domain reflectometry method

The dielectric behavior of native and heat-denatured ovalbumins (OVAs) from three avian species in aqueous solution was examined over a frequency range of 100 kHz to 20 GHz, using the time domain reflectometry (TDR) method. For the native OVA solutions, three kinds of relaxation processes were observed at around 10 MHz, 100 MHz, and 20 GHz, respectively; these could be assigned to the overall rotation of protein molecules, the reorientations of the bound water, and the free water molecules, respectively. For the heat-denatured samples, three relaxation processes were also observed. However, the relaxation process at approximately 100 MHz originated via a different mechanism other than the reorientation of bound water, namely, the micro-Brownian motion of peptide chains of heat-denatured protein. From the observed relaxation process at approximately 100 MHz, the relaxation strength of heat-denatured OVA solution for duck was higher than that of OVA solutions for hen and guinea fowl and showed the pH dependency from pH 7.0 to 8.0 for OVAs obtained from all three species. Furthermore, the results demonstrated that the relaxation strength was closely related to surface hydrophobicity of protein molecules and gel rheological properties. It was suggested that the difference in the surface hydrophobicity of protein influenced the dielectric behavior of water around denatured protein, whereas the dielectric behavior of denatured protein could be an indication of the gel rheological properties. Such studies can aid in the understanding of the different network structures of OVA gels from three avian species.     

横跨中国崇明岛河岸湿地缓冲区土壤中溶解性有机碳的空间分布及其荧光特征

The migration of soil dissolved organic carbon(DOC) from terrestrial to aquatic environments has important impacts on the adjacent water quality and the transport of organic and inorganic contaminants.However,few studies have investigated the sources and properties of DOC in riparian zones.A total of 34 soil samples were collected across four riparian buffer zones(Zones A-D) on Chongming Island,China.The vertical distributions of soil organic carbon(SOC) and DOC,fluorescence excitation-emission matrix(EEM) spectra of DOC and the optical indices,including fluorescence index(FI),index of recent autochthonous contribution(BIX),and humification index(HIX),were measured across the riparian environment to investigate the sources and fluorescence properties of DOC.The results showed that SOC stored in the surface soil(0-30 cm) accounted for 40%of the total soil profile SOC.The DOC accumulated in Zones A-C,which accounted for 5%of the SOC.The fluorescence EEM spectra of DOC showed that DOC contained humic-like and protein-like substances,which were mainly derived from recent plant debris by microbes.A large amount of humic-like substances were sorbed to minerals in the surface soil(0-30 cm).In addition,the riparian topography and soil physico-chemical properties(pH,EC and moisture) dictated the transformation and transport of DOC.The results suggested that EEMs could reveal the source of DOC in riparian soil systems,and that optical indices were complementary tools that revealed the characteristics of soil DOC and provided supplemental evidence about DOC sources.     

Kinetics of formation and functional properties of conjugates prepared by dry-state incubation of beta-lactoglobulin/acacia gum electrostatic complexes

The formation of conjugates between beta-lactoglobulin and acacia gum based on electrostatic complexes formed at pH 4.2 was investigated upon dry-state incubation for up to 14 days at 60 degrees C and 79% relative humidity (RH). By means of SEC-HPLC and RP-HPLC, it was shown that the beta-lactoglobulin incubated alone was able to form polymers with molecular masses higher than 200 kDa until 50% of the initial monomeric protein disappeared after 14 days. In the presence of acacia gum at initial protein to polysaccharide weight mixing ratios of 2:1 and 1:2, only 35% of the initial beta-lactoglobulin monomers disappeared after 14 days. Using RP-HPLC, an apparent reaction order of 2 was found for the disappearance of monomeric beta-lactoglobulin both in the presence or absence of acacia gum. However, the reaction rate was faster in the absence of acacia gum. SDS-PAGE electrophoresis with silver staining confirmed the formation of beta-lactoglobulin/acacia gum conjugates. The solubility curves of the incubated beta-lactoglobulin showed a minimum around pH 4-5. By contrast, the minimum of solubility of the beta-lactoglobulin/acacia gum incubated mixtures shifted to lower pH values compared to initial mixtures. The conjugates exhibited higher foam capacity than the incubated protein as well as lower equilibrium air/water surface tension. Conjugation at ratio 1:2 led to increased interfacial viscosity (300 mN s m(-1) at 0.01 Hz) compared to beta-lactoglobulin alone (100 mN s m(-1) at 0.01 Hz), but similar interfacial elasticity (30-40 mN m(-1)). The foam capacity of the conjugates was significantly higher than that of the incubated beta-lactoglobulin as well as foam expansion and drainage time, especially at pH 5.3, i.e., higher than the pH of formation of the conjugates.     

Use of catalyst in a 3D-QSAR study of the interactions between flavor compounds and beta-lactoglobulin

This paper reports a 3D-QSAR study using Catalyst software to explain the nature of interactions between flavor compounds and beta-lactoglobulin. A set of 35 compounds, for which dissociation constants were previously determined by affinity chromatography, was chosen. The set was divided into three subsets. An automated hypothesis generation, using HypoGen software, produced a model that made a valuable estimation of affinity and provided an explanation for the lack of correlation previously observed between the hydrophobicity of terpenes and the affinity for the protein. On the basis of these results, it appears that aroma binding to beta-lactoglobulin is caused by both hydrophobic interactions and hydrogen bonding, which plays a critical role. Catalyst appears to be a reliable tool for the application of 3D-QSAR study in aroma research.     

Structure and hydraulic properties of the boreal clay soil under differently managed buffer zones

Vegetated buffer zones (BZs) between arable fields and bodies of water are commonly established to reduce erosion and run‐off of particle‐bound nutrients. Functioning of a BZ depends on soil structure, as it is important for water infiltration. Therefore, it is vital to understand how varying management practices affect soils of BZs. We studied the structural and hydraulic properties of three differently managed BZs established in a boreal Vertic Stagnic Cambisol (clay, 51%). The three management practices for vegetation were as follows: natural with no treatment, harvested yearly and grazed by cattle. We used bulk density and macroporosity, together with a pore geometry index (air permeability per unit air‐filled porosity), to describe the soil structural properties. Hydraulic properties were measured at different length scales by means of an aggregate sorptivity test, saturated hydraulic conductivity of the core samples and field‐saturated hydraulic conductivity. Vegetation management markedly affected the physical properties in the top 5 cm of the soil. Properties were least favourable for infiltration at the grazed site, with the greatest bulk density, least macroporosity and hydraulic conductivity or greatest pore tortuosity. In general, spatial variation in zones with restricted and good hydraulic conductivity together with reduced aggregate sorptivity in the deeper horizons made the soil prone to preferential flow when initially dry. Prolonged wetness, on the other hand, reduced saturated hydraulic conductivity significantly, resulting in surface run‐off. Harvesting was considered the best management practice due to its inherent capacity for reducing the soil nutrient content and because it has minor implications for soil physical properties.     

Effect of sulfated polysaccharides on heat-induced structural changes in beta-lactoglobulin

The mechanism that leads to a decreased aggregation of beta-lactoglobulin in the presence of dextran sulfate and lambda-carrageenan was investigated by assessing changes in the denaturation thermodynamics and protein structure. Differential scanning calorimetry results showed that the denaturation temperature (Tp) was about 4.6 degrees C higher in the presence of dextran sulfate, as compared with beta-lactoglobulin alone, whereas in the presence of lambda-carrageenan the difference in Tp was about 1.2 degrees C. Changes in protein structure studies using near-UV circular dichroism (CD) provided support for the calorimetric results. The transition midpoint (Tm) for denaturation of beta-lactoglobulin was about 5 degrees C higher in the presence of dextran sulfate than that found with beta-lactoglobulin alone and about 2 degrees C in the presence of lambda-carrageenan. Thermal modifications of the tertiary structure of beta-lactoglobulin were irreversible at temperatures above 67 degrees C; the addition of dextran sulfate reduced the extent of such modifications. Far-UV CD studies indicated that the addition of dextran sulfate or lambda-carrageenan did not affect secondary structure changes of beta-lactoglobulin upon heating. These studies indicate that dextran sulfate and lambda-carrageenan can enhance the stability of beta-lactoglobulin and thereby inhibit heat denaturation and aggregation.     

Antioxidant mechanisms of enzymatic hydrolysates of beta-lactoglobulin in food lipid dispersions

The antioxidant activities of aqueous phase beta-lactoglobulin (beta-Lg) and its chymotryptic hydrolysates (CTH) were compared in this study. Proteins and peptides have been shown to inhibit lipid oxidation reactions in oil-in-water emulsions; however, a more fundamental understanding of the antioxidant activity of these compounds in dispersed food lipid systems is lacking. CTH was more effective than an equivalent concentration of beta-Lg in retarding lipid oxidation reactions when dispersed in the continuous phase of Brij-stabilized oil-in-water emulsions (pH 7). Furthermore, it was observed that CTH had higher peroxyl radical scavenging and iron-binding values than beta-Lg. Liquid chromatography-mass spectrometry (LC-MS) was used to measure the rate of oxidation of three oxidatively labile amino acid residues (Tyr, Met, and Phe) in certain CTH peptide fragments. Significant oxidation of specific Tyr and Met residues present in two separate 12 amino acid peptide fragments was observed in the days preceding lipid oxidation (39 and 55% of Tyr and Met were oxidized, respectively, by day 4 of the study); however, no significant oxidation of the Phe residue present in a specific 14 amino acid peptide fragment could be observed during the same time period. These data could suggest that Met and Tyr residues are capable of scavenging radical species and have the potential to improve the oxidative stability dispersed food lipids.     

中亚热带四种森林土壤团聚体及其有机碳分布特征

选择中亚热带地区4种典型森林类型:杉木林、湿地松林、毛竹林和次生林4种森林土壤为研究对象,研究了森林类型对土壤不同发生层水稳性团聚体及其有机碳分布特征的影响。结果表明:不同森林类型对>5 mm和2~5 mm土壤团聚体含量影响显著(p<0.05),表现为次生林>杉木林>毛竹林>湿地松林,而在其他粒径无显著差异。0~30 cm土层内团聚体R0.25和MWD次生林显著高于其他人工林,杉木林次之,湿地松林和毛竹林最低,其他土层无显著差异。各森林类型同土层不同粒径团聚体中有机碳含量随粒径大小变化,团聚体粒径越小,有机碳含量越高。0~10 cm土层同粒径土壤团聚体有机碳含量从大到小依次是次生林、杉木林、湿地松林和毛竹林,而在其他土层各森林类型之间差异不显著。