Interactions of soluble peptides and proteins from skeletal muscle on the release of volatile compounds

The ability of skeletal dipeptides (carnosine and anserine) and a sarcoplasmic protein (myoglobin) to interact with key flavor compounds (hexanal, octanal, methional, 2-pentanone, 2-methylbutanal, and 3-methylbutanal) has been studied using the solid phase microextraction (SPME) technique. Conditions for SPME analysis (fiber coating, sampling time, and linearity of detection) were optimized. The effect of pH on the binding was also investigated. Thermodynamic models were applied to evaluate the binding parameters n (number of binding sites), K (affinity constant), and DeltaG (Gibb's free energy) to all of the flavor compounds studied, and they showed an absence of cooperative effect. Carnosine was the peptide with the highest affinity for all of the volatile compounds except 2-pentanone. Its interaction with hexanal and methional was significantly affected by pH. Anserine showed a lower level of interactions with hexanal, methional, 2-methylbutanal, and 3-methylbutanal, whereas myoglobin interacted with only hexanal and 2-methylbutanal. Differences in aroma retention can thus result in different sensory perceptions of muscle foods.     

Autoxidation of packed almonds as affected by maillard reaction volatile compounds derived from roasting

Shelled almonds of two Italian varieties, Romana and Pizzuta, peeled and unpeeled, were roasted and packed under different conditions: air (control), vacuum, and Maillard reaction volatile compounds (MRVc) derived from the roasting process. Samples were stored for approximately 8 months at room temperature, without light, and, at regular intervals, were collected and analyzed to evaluate the progress of lipid oxidation. Peroxide values, triglyceride oligopolymers, and oxidized triglycerides were evaluated during the storage time. Results showed that, although the MRVc atmosphere did not protect the lipid fraction of almonds as well as the vacuum condition; nevertheless, it was more protective than the control atmosphere, showing an antioxidant effect. The effect of the natural coating was a strong protection against lipid oxidation; in fact, only the unpeeled samples showed peroxide values lower than the threshold of acceptability (25 milliequiv of O(2)/kg of oil). Moreover, at the end of the storage period, Pizzuta almonds showed a greater deterioration than those of the Romana variety.     

Furan formation from lipids in starch-based model systems, as influenced by interactions with antioxidants and proteins

The formation of furan upon sterilization of a lipid-containing starch gel was investigated in the presence of various antioxidants, namely, α-tocopherol, β-carotene, and ascorbic acid, with and without proteins. Results indicated that α-tocopherol did not significantly influence furan formation from oxidized lipids. β-Carotene, suggested previously to be a furan precursor itself, did influence the generation of furan in a concentration-dependent manner, although to a limited extent. Surprisingly, the presence of lipids seemed to limit the furan generation from β-carotene. Interestingly, the addition of ascorbic acid to the emulsions containing soybean or sunflower oils considerably enhanced the formation of furan from these oils. This was also the case when fresh oils were applied, shown previously to be nearly unable to generate furan. This observation can be explained by an intensified ascorbic acid degradation stimulated by the presence of lipids.     

Myoglobin oxidation in a model system as affected by nonheme iron and iron chelating agents

A model system was used to study the effect of nonheme iron on myoglobin oxidation at pH 5.6 and pH 7.2 at 23 degrees C. The addition of ferrous iron significantly (p < 0.05) increased the rate of myoglobin oxidation in the absence of lipid, demonstrating that iron promoted myoglobin oxidation independent of the effect of lipid oxidation. The addition of the type II, iron chelating antioxidants sodium tripolyphosphate (at pH 7.2) or milk mineral (at pH 5.6) negated the effect of added iron, slowing oxidation of myoglobin. A clear concentration dependence was seen for iron-stimulated myoglobin oxidation, based on both spectral and visual evidence. Further investigation is needed to determine the possible role for nonheme ferrous iron on myoglobin oxidation in vivo or in meat.     

Formation of odorants in Maillard model systems based on l-proline as affected by pH

Formation of the odorants acetic acid, 4-hydroxy-2,5-dimethyl-3-(2H)-furanone (HDMF), 6-acetyl-1,2,3,4-tetrahydropyridine (ATHP), and 2-acetyl-1-pyrroline (AP) was monitored by isotope dilution assays at pH 6, 7, and 8 in Maillard model reactions containing glucose and proline (Glc/Pro) or the corresponding Amadori compound fructosyl-proline (Fru-Pro). In general, higher yields were obtained at pH 7 and 8. Acetic acid was the major odorant with up to 40 mg/mmol precursor followed by HDMF (up to 0.25 mg/mmol), the formation of which was favored in the Fru-Pro reaction systems. On the contrary, ATHP (up to 50 microg/mmol) and AP (up to 5 microg/mmol) were more abundant in Glc/Pro. However, the sensory relevance of the two N-heterocycles was more pronounced on the basis of odor activity values, confirming their contribution to the overall roasty note of the reaction samples. It was also found that formation and decomposition of Fru-Pro were faster at pH 7 as compared to pH 6, explaining in part the preferred formation of the four odorants studied under neutral and slightly alkaline conditions. After 4 h of reaction at pH 7 in the presence of proline, about one-fourth of the glucose was consumed leading to acetic acid with a transformation yield of almost 40 mol %.     

Oxidation of skeletal muscle myofibrillar proteins in oil-in-water emulsions: interaction with lipids and effect of selected phenolic compounds

The effect of selected phenolic compounds, namely, gallic acid, cyanidin-3-glucoside, (+)-epicatechin, chlorogenic acid, genistein and rutin (50 and 200 microM), and alpha-tocopherol (50 microM) against the oxidation of oil-in-water emulsions (37 degrees C/10 days) containing 1% myofibrillar proteins (MPs), was investigated. Emulsions containing 1% bovine serum albumin (BSA) were also prepared for comparative purposes. Protein oxidation was assessed by measuring the loss of natural tryptophan fluorescence and the protein carbonyl gain by using fluorescence spectroscopy. Lipid oxidation was concurrently analyzed by measuring the increase of conjugated dienes (CDs) and hexanal. Proteins inhibited lipid oxidation in oil-in-water emulsions, and MPs showed a more intense antioxidant activity than BSA. MPs were also more resistant to oxidative deterioration than BSA. The different antioxidant capacity of MPs and BSA and their susceptibility to suffer oxidative reactions might be derived from their different amino acid composition and three-dimensional structures. The addition of the phenolic compounds resulted in a variety of effects, including both antioxidant and pro-oxidant effects. Gallic acid, cyanidin-3-glucoside, and genistein were the most efficient inhibitors of lipid and protein oxidation. The chemical structure of the phenolic compounds as well as the nature and conformation of the proteins were greatly influential on the overall effect against oxidative reactions.     

Influence of lambda-carrageenan on the release of systematic series of volatile flavor compounds from viscous food model systems

The effect of lambda-carrageenan addition level (0.1, 0.25, 0.4, and 0.5% w/w) and viscosity on the release of systematic series of aroma compounds (aldehydes, esters, ketones, and alcohols) was studied in thickened viscous solutions containing lambda-carrageenan and 10 wt % of sucrose. Air-liquid partition coefficients K (37 degrees C) of a total of 43 aroma compounds were determined in pure water and in the lambda-carrageenan solutions by static headspace gas chromatography. Mass transfer of the aroma compounds in water and in the thickened lambda-carrageenan solutions which had a wide viscosity range was assessed by dynamic headspace gas chromatography. K (37 degrees C) increased as the carbon chain increased within each homologous series. Esters exhibited the highest volatility, followed by aldehydes, ketones, and alcohols. Under equilibrium, no overall effect of lambda-carrageenan was found, except with the most hydrophobic compounds. Analysis of flavor release under nonequilibrium conditions revealed a suppressing effect of lambda-carrageenan on the release rates of aroma compounds, and the extent of decrease in release rates was dependent on the physicochemical characteristics of the aroma compounds, with the largest effect for the most volatile compounds. However, none of the effects was of a magnitude similar to the obtained changes in the macroscopic viscosity, and the suppressing effects are therefore attributable to the thickener and not the physical properties of the increasingly viscous systems.     

Bioavailability of allelochemicals as affected by companion compounds in soil matrices

Multicompound allelochemical interactions were studied using Centaurea maculosa as a model source to understand how the bioavailability of complex allelochemical mixtures is modified in soil-microbial systems. Litter decomposition of C. maculosa in sandy loam soil yielded five phenolic acids, namely, hydroxybenzoic, vanillic, protocatechuic, p-coumaric, and ferulic acids. The degradation studies were conducted by exogenous application of catechin, the primary allelochemical exuded by C. maculosa, and the phenolic acid cosolutes in a sandy loam and silt loam soil. Compared to a single-solute system, in a multisolute system the persistence of individual allelochemicals was significantly increased in both soils. Oxidation and sorption were primarily involved in the disappearance of allelochemicals. Mass spectrometric data showed that catechin rapidly underwent polymerization to form procyanidin dimer both in soil and in bioassay medium, resulting in reduced persistence and phytotoxicity. Hence, catechin phytotoxicity could occur only under conditions that would inhibit these condensation reactions. This study clearly demonstrates that various soil mechanisms including competitive sorption and preferential degradation would increase the persistence of allelochemical mixtures in a soil matrix.     

In vitro proteolysis of myofibrillar proteins from beef skeletal muscle by caspase-3 and caspase-6

The objective of the study was to investigate in vitro degradation of myofibrils by caspase-3 or -6. Myofibrillar proteins prepared from beef skeletal muscle were incubated with caspase-3 or -6 at 30 °C for 2 or 12 h, and subsequently, protein degradation was detected. Results showed that caspase-3 and -6 reproduced the degradation patterns of titin and nebulin observed during normal postmortem (PM) aging; however, they only reproduced the 28 kDa fragment derived from troponin-T. Caspase-3 induced only minor degradation of desmin. However, caspase-6 caused increasing degradation of desmin with extended incubation time and produced three degradation fragments (45, 29, and 27 kDa) of which only the 45 kDa fragment has been reported in aged beef. Therefore, caspase-3 or -6 could only reproduce a part of myofibrillar protein degradation or degradation fragments observed in naturally aged meat and may be involved in PM proteolysis of muscle proteins together with other endogenous proteases.     

Dynamic headspace analysis of the release of volatile organic compounds from ethanolic systems by direct APCI-MS

Static equilibrium headspace was diluted with a stream of nitrogen to study the stability of the volatile headspace concentration. The headspace dilution profile of 18 volatile compounds above aqueous and ethanolic solutions was measured in real time using atmospheric pressure chemical ionization-mass spectrometry. Under dynamic conditions the volatiles headspace concentration above water solutions decreased readily upon dilution. The presence of ethanol helped to maintain the volatile headspace concentration when the ethanol solution concentration was above 50 mL/L. This effect was such that under dynamic conditions the absolute volatile concentration above an ethanolic solution was higher than that above an aqueous solution, contrary to results observed in equilibrium studies. The ratio of the headspace concentration of volatiles above ethanolic 120 mL/L and water solutions was correlated to their air/water partition coefficient.     

Solubility improvement of shellfish muscle proteins by reaction with glucose and its soluble state in low-ionic-strength medium

When myofibrillar proteins of scallop striated adductor muscle were reacted with glucose through the Maillard reaction, the change in the solubility of myofibrillar proteins in 0.05-0.5 M NaCl solutions during glycosylation and their soluble states were investigated. The solubility in low-ionic-strength media increased greatly with the progress of the Maillard reaction. The solubility in 0.1 M NaCl reached 83% when more than 60% of lysine residues in myofibrillar proteins were modified by glucose. However, the excess progress of the Maillard reaction impaired the improved solubility of myofibrillar proteins in a low-ionic-strength medium. Myosin, actin, and paramyosin in glycosylated myofibrillar proteins were solubilized independently regardless of NaCl concentration. In addition, the glycosylated myosin lost its filament-forming ability and existed as a monomer in 0.1 M NaCl.     

Potassium budgets in grassland systems as affected by nitrogen and drainage

Abstract. The main inputs, outputs and transfers of potassium (K) in soils and swards under typical south west England conditions were determined during 1999/00 and 2000/01 to establish soil and field gate K budgets under different fertilizer nitrogen (N) (0 and 280 kg ha⁻¹ yr⁻¹) and drainage (undrained and drained) treatments. Plots receiving fertilizer N also received farmyard manure (FYM). Potassium soil budgets ranged, on average for the two years, from −5 (+N, drained) to +9 (no N and undrained) kg K ha⁻¹ yr⁻¹ and field gate budgets from +23 (+N, drained) to +89 (+N, undrained). The main inputs and outputs to the soil K budgets were fertilizer application (65%) and plant uptake (93%). Animals had a minor effect on K export but a major impact on K recycling. Nitrogen fertilizer application and drainage increased K uptake by the grass and, with it, the efficiency of K used. It also depleted easily available soil K, which could be associated with smaller K losses by leaching.     

Arylamidase and amidohydrolases in soils as affected by liming and tillage systems

This study investigated the long-term effect of lime application and tillage systems (no-till, ridge-till and chisel plow) on the activities of arylamidase and amidohydrolases involved in N cycling in soils at four long-term research sites in Iowa, USA. The activities of the following enzymes were assayed: arylamidase, -asparaginase, -glutaminase, amidase, urease, and -aspartase at their optimal pH values. The activities of the enzymes were significantly (P<0.001) and positively correlated with soil pH, with r values ranging from 0.42^* to 0.99^*** for arylamidase, 0.81^*** to 0.97^*** for -asparaginase, 0.62^*** to 0.97^*** for -glutaminase, 0.61^*** to 0.98^*** for amidase, 0.66^** to 0.96^*** for urease, and 0.80^*** to 0.99^*** for -aspartase. The Δactivity/ΔpH values were calculated to assess the sensitivity of the enzymes to changes in soil pH. The order of the sensitivity of enzymes was as follows: -

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-aspartase. The enzyme activities were greater in the samples of the 0–5 cm depth than those of the 0–15 cm samples under no-till treatment. Most of the enzyme activities were significantly (P<0.001) and positively correlated with microbial biomass C (C_mic) and N (N_mic). Lime application significantly affected the specific activities of the six enzymes studied. Results showed that soil management practices, including liming and type of tillage significantly affect soil biological and biochemical properties, which may lead to changes in nitrogen cycling, including N mineralization in soils.     

Photosynthesis,chlorophyll fluorescence and soluble carbohydrates in sunflower leaves as affected by boron deficiency

The effect of various boron levels in the nutrient solution on the growth, boron and chlorophyll content, photosynthesis and chlorophyll fluorescence in the leaves of young sunflower (Helianthus annuus L.) plants was studied under greenhouse conditions. Deficiency of boron decreased the dry matter yield of the roots, shoots and leaves. The content of boron in all analyzed plant parts increased with the increase of boron levels in the nutrient solution, more so in the shoots than in the roots. Leaf area was reduced under boron deficiency as well as the content of chlorophyll in the leaves. The content of analyzed sugars was increased in boron deficient plants, glucose content exhibited the highest increase under boron deficiency. Boron deficiency appreciably decreased photosynthetic oxygen evolution by leaves, the apparent quantum yield and quantum efficiency of photosystem two electron transport. The diminished rate of photosynthesis in boron deficient sunflower leaves could be correlated to the diminished efficiency of electron transport and to the increased content of sugars in the leaves.     

Urease activity of microbial biomass in soils as affected by cropping systems

 The impacts of crop rotations and N fertilization on different pools of urease activity were studied in soils of two long-term field experiments in Iowa; at the Northeast Research Center (NERC) and the Clarion-Webster Research Center (CWRC). Surface soil samples (0–15 cm) were taken in 1996 and 1997 in corn, soybeans, oats, or meadow (alfalfa) plots that received 0 or 180 kg N ha^–1, applied as urea before corn and an annual application of 20 kg P and 56 kg K ha^–1. The urease activity in the soils was assayed at optimal pH (THAM buffer, pH 9.0), with and without toluene treatment, in a chloroform-fumigated sample and its nonfumigated counterpart. The microbial biomass C (C_mic) and N (N_mic) were determined by chloroform fumigation methods. The total, intracellular, extracellular and specific urease activities in the soils of the NERC site were significantly affected by crop rotation, but not by N fertilization. Generally, the highest total urease activities were obtained in soils under 4-year oats–meadow rotations and the lowest under continuous corn. The higher total activities under multicropping systems were caused by a higher activity of both the intracellular and extracellular urease fractions. In contrast, the highest values for the specific urease activity, i.e. of urease activity of the microbial biomass, were found in soils under continuous soybean and the least under the 4-year rotations. Total and extracellular urease activities were significantly correlated with C_mic (r>0.30* and >0.40**) and N_mic (r>0.39** and >0.44**) in soils of the NERC and CWRC sites, respectively. Total urease activity was significantly correlated with the intracellular activity (r>0.73***). About 46% of the total urease activity of the soils was associated with the microbial biomass, and 54% was extracellular in nature. Received: 25 May 1999     

Arylsulfatase activity of microbial biomass in soils as affected by cropping systems

 The impacts of crop rotations and N fertilization on different pools of arylsulfatase activity (total, intracellular, and extracellular) were studied in soils of two long-term field experiments in Iowa to assess the contibution of the microbial biomass to the activity of this enzyme. Surface-soil samples were taken in 1996 and 1997 in corn, soybeans, oats, or meadow (alfalfa) plots that received 0 or 180 kg N ha^–1 before corn, and an annual application of 20 kg P ha^–1 and 56 kg K ha^–1. The arylsulfatase activity in the soils was assayed at optimal pH (acetate buffer, pH 5.8) before and after chloroform fumigation; microbial biomass C (C_mic) and N (N_mic) were determined by chloroform-fumigation methods. All pools of arylsulfatase activity in soils were significantly affected by crop rotation and plant cover at sampling time, but not by N fertilization. Generally, the highest total, intracellular, and extracellular arylsulfatase activities were obtained in soils under cereal-meadow rotations, taken under oats or meadow, and the lowest under continuous cropping systems.Total, intracellular, and extracellular arylsulfatase activities were significantly correlated with C_mic (r>0.41, P<0.01) and N_mic (r>0.38, P<0.01) in soils. The averages of specific activity values, i.e., of arylsulfatase activity of the microbial biomass, expressed per milligram C_mic, ranged from 315 to 407 μg p-nitrophenol h^–1. The total arylsulfatase activity was significantly correlated with the intracellular activity, with r values >0.79 (P<0.001). In general, about 45% of the total arylsulfatase activity was extracellular, and 55% was associated with the microbial biomass in soils, indicating the importance of the microflora as an enzyme source in soils. Received: 23 April 1998     

Kinetics of interaction of vanillin with amino acids and peptides in model systems

Model systems were used to study the reaction kinetics of vanillin and pentalysine, lysine, glutathione, cysteine, aspartame, or phenylalanine (molar ratio 1:1) in phosphate buffer. The buffer pH was adjusted to the pK(a)(2) of the available alpha-amino group of each amino acid or peptide. Reductions of vanillin followed first-order kinetics at 55, 65, and 75 degrees C in the presence of each of the amino acids or peptides used. The reaction rates were accelerated as the temperature increased. The rate constants were highest for pentalysine followed by lysine, phenylalanine, glutathione/cysteine, and aspartame. The reduction of phenylalanine followed first-order kinetics, whereas the formation of its reaction product followed zero-order kinetics. The activation energy (E(a)) for the reaction ranged from 5.6 to 14.5 kcal/mol.     

Effect of pH, temperature, and moisture on the formation of volatile compounds in glycine/glucose model systems

Mixtures of glycine, glucose, and starch were extrusion cooked using sodium hydroxide at 0, 3, and 6 g/L of extruder water feed, 18% moisture, and 120, 150, and 180 degrees C target die temperatures, giving extrudates with pH values of 5.6, 6.8, and 7.4. Freeze-dried equimolar solutions of glucose and glycine were heated either dry or after equilibration to approximately 13% moisture at 180 degrees C in a reaction-tube system designed to mimic the heating profile in an extruder. Volatile compounds were isolated onto Tenax and analyzed by gas chromatography-mass spectrometry. For the extrudates, total yields of volatiles increased with decreasing pH at 180 degrees C, reached a maximum at pH 6.8 at 150 degrees C, and increased with increasing pH at 120 degrees C. Amounts increased with temperature at all pH values. Pyrazines were the most abundant class for all sets of conditions (54-79% of total volatiles). Pyrroles, ketones, furans, oxazoles, and pyridines were also identified. Yields of volatiles from the reaction-tube samples increased by >60% in the moist system. Levels of individual classes also increased in the presence of moisture, except pyrazines, which decreased approximately 3.5-fold. Twenty-one of the compounds were common to the reaction-tube samples and the extrudates.     

Soil compressibility and penetrability of an Oxisol from southern Brazil, as affected by long-term tillage systems

The precompression stress value defines the transition from the reloading curve to the virgin compression line in the stress–strain curve, which can be used to quantify the highest load or the most intense predrying previously applied to the soil. Thus, in soils with well-defined structured soil horizons, each layer can be characterized by such mechanical strength. Penetration resistance measurements, on the other hand, can be used to determine total soil strength profiles in the field. The effect of long-term tillage systems on physical and mechanical properties was determined in undisturbed and remolded samples collected at 5 and 15 cm depth, 6 months after applying no-till (NT), chisel plow (CP), and conventional tillage (CT) treatments, along with the application of mineral fertilizer and poultry litter. The compressibility tests were performed under confined conditions, with normal loads varying from 10 to 400 kPa after a defined predrying to −6 or −30 kPa. Penetration resistance was determined in the field, after seeding, in three positions: seeding row (SR), untrafficked interrow (UI), and recently trafficked interrow (TI). No-till system showed greater soil resistance to deformation than tilled treatments, as determined by the higher precompression stress and lower coefficient of compressibility. When original soil structure was destroyed (remolded samples), smaller differences were found. The application of extra organic matter (poultry litter) resulted in a reduction of precompression stress in undisturbed samples. Penetration resistance profiles showed greater differences among tillage treatments in the upper layer of the untrafficked interrow, where NT system showed the higher values. Smaller differences were found in the seeding row (with lower values) and in recently trafficked interrow (with higher values), showing that even traffic with a light tractor after soil tillage reduced drastically the effect of previous tillage by loosening up the soil. On the other hand, the tool used to cut the soil and to open the furrow for seeding, incorporated in the direct seeding machine, was sufficient to realleviate surface soil compaction.     

Changes of folates, dietary fiber, and proteins in wheat as affected by germination

Wheat kernels of the cultivar 'Tommi' were germinated for up to 168 h at 15, 20, 25, or 30 degrees C. Samples were taken at different stages of germination and were analyzed for the quantitative protein composition using an extraction/HPLC method, for folate vitamers using a stable isotope dilution assay, and for soluble, insoluble, and total dietary fiber using a gravimetric method. Gluten proteins were substantially degraded during germination. During the first stages of germination the degradation of glutenins was predominant, whereas longer germination times were required to degrade gliadins. The optimal temperature for gliadin degradation was 20 degrees C, and that for glutenin degradation was 25 degrees C. Omega5- and omega1,2-gliadins were less sensitive to proteolytic degradation than alpha- and gamma-gliadins, and LMW subunits of glutenin were less sensitive than HMW subunits. During germination a time- and temperature-dependent increase of total folate occurred. A maximum 3.6-fold concentration was obtained after 102 h of germination at 20 and 25 degrees C including 5-methyltetrafolate as the major vitamer. The concentration of dietary fiber remained constant or decreased during the first 96 h of germination. Prolonged germination times of up to 168 h led to a substantial increase of total dietary fiber and to a strong increase of the soluble dietary fiber by a factor of 3, whereas the insoluble fiber decreased by 50%.