Heat-induced gelation of chicken Pectoralis major myosin and beta-lactoglobulin

The denaturation, aggregation, and rheological properties of chicken breast muscle myosin, beta-lactoglobulin (beta-LG), and mixed myosin/beta-LG solutions were studied in 0.6 M NaCl, 0.05 mM sodium phosphate buffer, pH 7.0, during heating. The endotherm of a mixture of myosin and beta-LG was identical to that expected if the endotherm of each protein was overlaid on the same axis. The maximum aggregation rate (AR(max)) increased, and the temperature at the AR(max) (T(max)) and initial aggregation temperature (T(o)) decreased as the concentration of both proteins was increased. The aggregation profile of <0.5% myosin was altered by the presence of 0.25% beta-LG. Addition of 0.5-3.0% beta-LG decreased storage moduli of 1% myosin between 55 and 75 degrees C, but increased storage moduli (G') when heated to 90 degrees C and after cooling. beta-LG had no effect on the gel point of > or =1.0% myosin, but enhanced gel strength when heated to 90 degrees C and after cooling. After cooling, the G' of 1% myosin/2%beta-LG gels was about 1.7 times greater than that of gels prepared from 2% myosin/1% beta-LG.     

Effect of combined heat and high-pressure treatments on the texture of chicken breast muscle (pectoralis fundus)

Commercially supplied chicken breast muscle was subjected to simultaneous heat and pressure treatments. Treatment conditions ranged from ambient temperature to 70 degrees C and from 0.1 to 800 MPa, respectively, in various combinations. Texture profile analysis (TPA) of the treated samples was performed to determine changes in muscle hardness. At treatment temperatures up to and including 50 degrees C, heat and pressure acted synergistically to increase muscle hardness. However, at 60 and 70 degrees C, hardness decreased following treatments in excess of 200 MPa. TPA was performed on extracted myofibrillar protein gels that after treatment under similar conditions revealed similar effects of heat and pressure. Differential scanning calorimetry analysis of whole muscle samples revealed that at ambient pressure the unfolding of myosin was completed at 60 degrees C, unlike actin, which completely denatured only above 70 degrees C. With simultaneous pressure treatment at >200 MPa, myosin and actin unfolded at 20 degrees C. Unfolding of myosin and actin could be induced in extracted myofibrillar protein with simultaneous treatment at 200 MPa and 40 degrees C. Electrophoretic analysis indicated high pressure/temperature regimens induced disulfide bonding between myosin chains.     

Gelation of whey protein concentrate-cassava starch in acidic conditions.

Whey protein concentrate (WPC)-cassava starch (CS) gels were prepared by heating WPC-CS dispersions (0-12.5% protein-0-4.2% starch, w/w; pH 3.75 and 4.2). Gels were characterized by measures of water-holding capacity (WHC), estimation of the relative size and/or density distribution of the gel particles, and light microscopy. Differential scanning calorimetry (DSC) of WPC-CS dispersions was also performed. Results show that CS increased the WHC of gels. Mixed gels presented separate zones of gelatinized starch and aggregated protein and a higher proportion of large/high-density particles. DSC assays showed that starch gelatinization preceded protein denaturation during heating. Starch gelatinization shifted to higher temperatures in dispersions containing WPC, due to the presence of whey proteins, lactose, and calcium.     

Polymerization and gelation of whey protein isolates at low pH using transglutaminase enzyme

Dynamic and steady shear rheology is used to examine the synthesis of low-pH (approximately 4) whey protein gels obtained through a two-step process. The first step involves cross-linking of whey proteins at pH 8 and 50 degrees C using transglutaminase enzyme, while the second step entails cold-set acidification of the resulting solution using glucono-delta-lactone (GDL) acid. During the first step, the sample undergoes enzyme-catalyzed epsilon-(gamma-glutamyl)lysine bond formation with a substantial increase in viscosity. Acidification in the second step using GDL acid leads to a rapid decrease in pH with a concomitant increase in the elastic (G') and viscous (G' ') moduli and formation of a gelled network. We examine the large strain behavior of the gel samples using a relatively new approach that entails plotting the product of elastic modulus and strain (G'gamma) as a function of increasing dynamic strain and looking for a maximum, which corresponds to the yield or fracture point. We find the enzyme-catalyzed gels to have significantly higher yield/fracture stress and strain compared to cold-set gels prepared without enzyme or conventional heat-set gels. In addition, the elastic modulus of the enzyme-catalyzed gel is also higher than its non-enzyme-treated counterpart. These results are discussed in terms of the gel microstructure and the role played by the enzyme-induced cross-links.     

alpha-Casein improves the gel properties of dried egg white

The effects of addition of alpha-casein (alpha-CN) to dried egg white (DEW) were investigated by measuring transparency, hardness, and water-holding capacity (WHC) of the heat-induced gels. A DEW concentration of 8% (w/w) was required for formation of a self-supporting gel following heating at 80 degrees C for 20 min at pH 7. Solutions of alpha-CN, even up to a protein concentration of 12% (w/w), did not gel under the same conditions. The addition of alpha-CN (0.5-4%) to 8% DEW caused the increase in gel hardness gels, as compared with DEW gels alone at a total amount of protein concentrations, and the mixed gels became transparent with the increase of added alpha-CN concentrations. The 10% mixed protein solutions of alpha-CN (3-6%) and DEW (4-7%) formed transparent gels, although each protein did not gel individually at their protein concentrations. Mixture with 2:8 mixing ratio of alpha-CN to DEW at a total protein concentration of 10% showed synergistic effects in improving DEW gel properties above pH 7 and below 25 mM NaCl. The improvements (hardness, transparency, and WHC) of DEW gel by alpha-CN seem to be caused mainly by the inhibition of alpha-CN against heat coagulation of DEW protein.     

Effect of electron beam irradiation and storage at 5 degrees C on thiobarbituric acid reactive substances and carbonyl contents in chicken breast meat infused with antioxidants and selected plant extracts

This study evaluated the effectiveness of synthetic and natural antioxidants, green tea, commercial grape seed extracts/combinations, and TBHQ, with varying concentrations of lipid oxidation of nonirradiated and irradiated chicken breast meats stored at 5 degrees C for 12 days. Fresh boneless and skinless chicken breast meats were vacuum-infused with varying concentrations of antioxidants: green tea, grape seed extracts alone/in combination, and TBHQ. The irradiation dosage was 3.0 kGy. Carbonyl values of raw chicken meat and thiobarbituric acid reactive substances (TBARS) values of raw and cooked chicken meat were determined for 0-12 days at 5 degrees C storage. TBARS values for 0-12 days of storage at 5 degrees C ranged from 1.21 to 7.3 and 1.22 to 8.51 mg malondialdehyde/100 g chicken for nonirradiated and irradiated raw chicken, respectively. TBARS values of cooked chicken ranged from 2.19 to 35.83 and 2.45 to 45.72 mg malondialdehyde/100 g chicken for nonirradiated and irradiated chicken, respectively. Irradiation increased TBARS values of both controls and plant extracts. The carbonyl content in meat lipid ranged from 1.7 to 2.9 and 1.7 to 4.41 micromol acetophenone/10 g of nonirradiated and irradiated chicken meat, respectively, and meat protein ranged from 1.4 to 2.07 and 1.41 to 2.72 micromol/10 g meat. Infusion of chicken meat with selected plant extracts is an effective method to minimize lipid oxidation and volatiles developments caused by irradiation.     

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.     

Environmental factors that affect the ability of amylose to contribute to retrogradation in gels made from rice flour

Retrogradation in rice is a trait that describes the hardening of cooked rice after storage or cooling, and it has significant implications for many consumers of rice, since many people cook rice in the morning and consume it several hours later or the next day. Tools to select against retrogradation in breeding programs are yet to be described. Here, we aim to determine the effect on retrogradation of storage time and temperature and the role of starch, protein, and lipids using gels made from Koshihikari grown in either Australia or Japan. Immediately after cooking, cooling from 60 to 40 degrees C had a minimal effect on firmness, but cooling to 20 degrees C led to significantly firmer gels. Storing the gels at low temperatures did not have an additional effect on the firmness as compared with storing the gels at 20, 40, or 60 degrees C. The removal of proteins led to significantly softer gels at all storage treatments but did not affect the change in firmness on cooling. The removal of lipids increased the rate of retrogradation and the firmness of gels significantly for all treatments. Koshihikari grown in Japan retrograded much less than Koshihikari grown in Australia. The amount of amylose that could be washed from gels made from Australian flour was much greater than for gels made from Japanese flour. After storage, only low molecular weight amylose chains were released from the gel and only after rewarming them to 60 degrees C. Despite the fact that flours from both origins were 18% amylose, the amount of long amylose chains that were complexed with lipids was much greater for the Japanese rice, and the unavailability of the complexed long amylose chains explained the lack of retrogradation in the Japanese rice. Once the long chains were released from amylose-lipid complexes, the Japanese rice retrograded. Thus, the environmental factor affecting retrogradation in this variety is type or amount of lipids synthesized, and the degree of retrogradation was determined by the availability of long chains of amylose.     

Proteomic analysis and differential expression in protein extracted from chicken with a varying growth rate and water-holding capacity

Chickens from a randomly bred genetic line were segregated into high and low growth rates and high and low water-holding capacities (WHCs). The objective of this study was to identify protein markers associated with slow and fast growth rates and low and high WHCs from water-soluble protein (WSP) and crude myofibrillar protein (CMP) extracts of chicken breast muscle. Proteins were fractionated using two-dimensional electrophoresis, and a total of 22 protein spots were selected, excised, and analyzed by in-gel tryptic digestion and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Proteins expressed in extracts from slow and fast growth rates and low and high WHCs included metabolic enzymes, such as creatine kinase, pyruvate kinase, triosephosphate isomerase, and ubiqitin; housekeeping proteins, such as heat shock protein; contractile proteins, such as myosin heavy chain; actin; and also MHC isoforms and actin isoforms. The mass spectra of 20 protein spots significantly matched (protein score >83; P < 0.05) an online database. In CMP, there were unique proteins that were present only in the fast-growth population: gi|118099530 , gi|20664362 , gi|71895043 , gi|114794125 , gi|297343122 , and gi|71895043 . This information identified protein markers associated with growth rate and water holding capacity. Some of those protein markers could be added to the chicken database.     

"Weak gel"-type rheological properties of aqueous dispersions of nonaggregated kappa-carrageenan helices

The rheological properties of kappa-carrageenan helices dispersed in an aqueous medium, which prevents aggregation of helices, were investigated. A dispersion of 1.5% w/w nonaggregated kappa-carrageenan helices exhibited gel-like dynamic mechanical spectra at 20 degrees C; that is, the storage modulus G' predominated over the loss modulus G' ' in the entire frequency range examined (0.5-100 rad/s). However, the observed slight frequency dependence of the moduli and the relatively large value of tan delta (= G' '/G' > 0.1) were typical of so-called weak gels. The magnitude of G' of the kappa-carrageenan weak gels was less than that of conventional gels formed by 0.15% w/w kappa-carrageenan in an aggregating condition at 20 degrees C. Under large deformation, enough for the conventional gels to rupture, the weak gel systems flowed but never ruptured, suggesting that the weak gel-type rheological properties of the kappa-carrageenan dispersions were due to a sufficiently long relaxation time of topological entanglements among double-helical conformers but not due to the formation of a three-dimensionally percolated permanent network.     

Mechanical and water-holding properties and microstructures of soy protein isolate emulsion gels induced by CaCl2, glucono-δ-lactone (GDL), and transglutaminase: influence of thermal treatments before and/or after emulsification

The mechanical properties, water-holding capacities (WHC), and microstructures of emulsion gels, induced by glucono-δ-lactone (GDL), CaCl(2), and microbial transglutaminase (MTGase) from unheated and heated soy protein isolate (SPI)-stabilized emulsions (at protein concentration 5%, w/v; oil volume fraction, 20%, w/v), were investigated and compared. The influence of thermal pretreatments (at 90 °C for 5 min) before and/or after emulsification was evaluated. Considerable differences in mechanical, water-holding, and microstructural properties were observed among various emulsion gels. The thermal pretreatment after emulsification increased the strength of the emulsion gels induced by GDL and CaCl(2), whereas in the case of MTGase, thermal pretreatments before and/or after emulsification on the contrary greatly inhibited gel network formation. The application of the enzyme coagulant exhibited much higher potential to form SPI-stabilized emulsion gels with higher mechanical strength than that of the other two coagulants. The WHC of the emulsion gels seemed to be not directly related to their gel network strength. Confocal laser scanning microscope analyses indicated that the network microstructure of the formed emulsion gels, mainly composed of aggregated protein-stabilized oil droplets and protein aggregate clumps, varied with the type of applied coagulants and emulsions. The differences in microstructure were basically consistent with the differences in mechanical properties of the gels. These results could provide valuable information for the formation of cold-set soy protein-stabilized emulsion gels.     

Resveratrol and a novel tyrosinase in Carignan grape juice

Polyphenol levels in wines are affected by the wine-making process. Resveratrol is one polyphenol which has been the subject of a commendable amount of recent research. In this work, we found that resveratrol is immediately degraded by tyrosinase. A novel tyrosinase was purified from Carignan grapes. The purification process included salting out and separation on a cation-exchange column, followed by gel filtration. Tyrosinase was purified in a homogeneous form by SDS-PAGE and was characterized: its specific activity toward 3-(3,4-dihydroxyphenyl)-L-alanine (DOPA) increased by a factor of 24 with an overall recovery of 3% of initial activity. The apparent molecular mass of the purified tyrosinase was 40 kDa as determined by SDS-PAGE, and 42 kDa as determined by gel filtration. Its activity was optimal at pH 6 and at 25 degrees C. The enzyme exhibited high activity toward phenylenediamine, epicatechin, pyrogallol, DOPA, and resveratrol. Tyrosinase activity was inhibited by KCN, thiourea, and SO(2). Resveratrol levels were stable following the removal of proteins from the juice, suggesting that early spraying of grapes with SO(2) is an important factor affecting the final amount of resveratrol in wine.     

Effects of high pressure on the myofibrillar proteins of cod and turkey muscle

When turkey breast muscle and isolated myofibrillar protein and myosin of cod or turkey (pH approximately 7) were subjected to pressures up to 800 MPa for 20 min, DSC and electrophoresis (SDS-PAGE) indicated that high pressure-induced denaturation of myosin led to the formation of structures that contained hydrogen bonds and were additionally stabilized by disulfide bonds. Disulfide bonds were also important in heat-induced myosin gels. Hardness of whole cod muscle, estimated by texture profile analysis, showed pressure-treated samples (400 MPa) to be harder than cooked (50 degrees C) or cooked and then pressure-treated or pressure-treated and then cooked samples, supporting the suggestion that pressure induces the formation of heat labile hydrogen-bonded structures while heat treatment gives rise to structures that are primarily stabilized by disulfide bonds and hydrophobic interactions. As expected, turkey myosin is more stable than that of cod; however, it seems their pressure-induced gelation mechanisms are similar.     

Effect of pH on the rheological and structural properties of gels of water-washed chicken-breast muscle at physiological ionic strength

Adjustment of pH from 6.4 to neutrality improved gelling ability and water-holding capacity of twice water-washed, minced chicken-breast muscle significantly at physiological ionic strength, at which the majority of the myofibrillar proteins, including myosin, are not soluble. A strain value of 2.2 was obtained at neutral pH. Myofibrils were the main components of the gel network at both pH 6.4 and 7.0; however, the myofibrillar distribution varied with the pH value. At pH 6.4, myofibrils formed a network of localized aggregates leaving large voids between, whereas at neutral pH, an evenly distributed network of myofibrils was formed. In addition, at neutral pH, a network of fine strands was found within the network of myofibrils. The network was much less developed at pH 6.4. The thin and thick filaments within each myofibrillar structure were disorganized at both pH values. The intramyofibrillar spaces were larger at neutral pH than at pH 6.4. It was proposed that adjustment of pH to neutrality increased electrostatic repulsion leading to a more even distribution of the myofibrillar proteins, a key factor responsible for the improved gel strength and water-holding capacity.     

Phospholipid/fatty acid-induced secondary structural change in beta-lactoglobulin during heat-induced gelation

Effects of phosphatidylcholine (PC) and the predominant fatty acids (FAs) in milk, butyrate, oleate, and palmitate, on secondary structural changes in beta-lactoglobulin (beta-LG) during heat-induced gelation were analyzed on the basis of circular dichroism (CD) spectra. Small-strain oscillatory measurements were carried out to characterize viscoelastic properties of the heat-induced gels. In the absence of added salt, PC and FAs induced helix formation of beta-LG on heating to 80 degrees C and increased the storage moduli (G') of heat-induced gels. In the presence of 500 mM NaCl, PC did not change the CD spectrum of beta-LG but decreased G'. In contrast, butyrate substantially unfolded beta-LG in 500 mM NaCl on heating, forming very elastic gels with increased G' values. Palmitate and oleate induced beta-LG gel formation at 25 degrees C without heating; heating to 80 degrees C almost completely unfolded beta-LG in 500 mM NaCl.     

Enzyme-aided modification of chicken-breast myofibril proteins: effect of laccase and transglutaminase on gelation and thermal stability

The effect of laccase and transglutaminase (TG) on cross-linking, gelation, and thermal stability of salt-soluble chicken-breast myofibril proteins was investigated at pH 6. Both enzymes modified the protein pattern detected by SDS-PAGE. Identification of proteins by peptide mass mapping showed that myosin heavy chain (MHC) and troponin T were the most affected proteins. These proteins faded or disappeared as a function of the incubation time with both enzymes on SDS-PAGE. The molecular weight of actin was not, however, affected by either enzyme. The effects that the enzymes had on the gel formation of chicken-breast myofibrils were studied in 0.35 and 0.60 M NaCl solutions at 3% protein content and a constant temperature of 40 degrees C by using a small deformation viscoelastic measurement. TG substantially increased the storage modulus (G') of 3% protein in 0.35 M NaCl. Without the enzymes, gelation was insignificant in 0.35 M NaCl. The increased solubility of the proteins at 0.60 M NaCl intensified gelation with TG. G' increased 32 and 64% at dosages of 10 and 100 nkat of TG, respectively. Also, laccase increased G' of the gel in 0.60 M salt concentration. However, a high laccase dosage decreased the magnitude of G' below the control level. Differential scanning calorimetric (DSC) measurements indicated slightly reduced myosin heat stability after TG pretreatment and increased actin heat stability with both enzymes. Maximum transition temperatures did not alter with either enzyme.     

精油对熟制鸡胸肉中产气荚膜梭菌抑制效果预测模型研究

为研究不同香辛料精油对熟制鸡胸肉中产气荚膜梭菌(C.perfringens)的影响,该文以肉桂精油、艾草精油和茴香精油为研究对象,对C.perfringens标准株(ATCC13124)和分离株(C1)抑菌效果,筛选出抑制最佳浓度,采用BP神经网络构建C.perfringens的生长/残存动力学模型,并以相关系数(R~2)和均方根误差(RMSE)评价模型精度,以期快速预测不同精油浓度条件对C.perfringens影响。结果表明:经肉桂精油处理后的ATCC13124和C1浓度最低,抑制效果最强;采用BP神经网络模型构建不同精油对熟制鸡胸肉中C.perfringens的预测模型,肉桂精油对ATCC13124和C1的R~2分别为0.963和0.976,RMSE分别为0.327和0.271CFU/g,预测效果最佳;利用验证集对模型鲁棒性进行验证,结果表明R~2均在0.917以上,RMSE在0.200～0.640 CFU/g之间,结果表明,BP神经网络模型可以较好的预测熟制鸡胸肉中产气夹膜梭菌的生长/残存情况;为肉类加工过程中控制C.perfringens提供理论依据。     

Acid-induced gelation of enzymatically modified, preheated whey proteins

Low-pH whey protein gels are formulated using a sequential protocol of heat treatment, enzyme incubation, and cold-set acidification. The heat-induced disulfide and enzyme-catalyzed epsilon-(gamma-glutamyl)lysine linkages, both at neutral pH, produce a polymerized protein solution. The molecular weights of these samples show an exponential increase with protein concentration. The additional enzyme-catalyzed cross-links cause little change in molecular weight from that of heat-treated samples at low protein concentrations, indicating predominant intramolecular cross-linking. Enzyme treatment at higher protein concentration however causes increase in molecular weight, possibly due to formation of intermolecular cross-links. Acidification of the polymerized protein solutions through glucono-delta-lactone acid leads to gel formation at pH 4. The elastic (G') and viscous (G' ') moduli of gels with and without enzyme treatment show similar frequency dependence, indicating comparable microstructures, consistent with all samples exhibiting similar fractal dimensions of approximately 2 obtained independently using rheology and confocal microscopy. A substantial increase in fracture strain and stress of the gel is achieved by enzyme treatment. However, the elastic modulus (G') is only slightly larger after enzyme treatment compared with heat-treated samples. These results indicate that factors responsible for fracture properties may not be apparent in the gel microstructure and linear viscoelastic properties.     

Gelling properties of heat-denatured beta-lactoglobulin aggregates in a high-salt buffer

Thermal denaturation, rheological, and microstructural properties of gels prepared from native beta-lactoglobulin (beta-LG) and preheated or heat-denatured beta-LG (HDLG) aggregates were compared. The HDLG was prepared by heating solutions of 4% beta-LG in deionized water, pH 7.0, at 80 degrees C for 30 min and then diluted to the desired concentration in 0.6 M NaCl and 0.05 M phosphate buffer at pH 6.0, 6.5, and 7.0. When reheated to 71 degrees C, HDLG formed a gel at a concentration of 2% protein. At pH 7.0, 3% HDLG gelled at 52.5 degrees C and had a storage modulus (G') of 2200 Pa after cooling. beta-LG (3%) in 0.6 M NaCl and 0.05 M phosphate buffer, pH 7.0, did not gel when heated to 71 degrees C. The gel point of 3% HDLG decreased by 10.5 degrees C and the G' did not change when the pH was decreased to 6.0. The HDLG gel microstructure was composed of strands and clumps of small globular aggregates in contrast to beta-LG gels, which contained a particulate network of compacted globules. The HDLG formed a gel at a lower concentration and lower temperature than beta-LG in the high-salt buffer, suggesting an application in meat systems or other food products prepared with salt and processed at temperatures of < or =71 degrees C.     

Effect of Moisture Content at Harvest and Degree of Milling (Based on Surface Lipid Content) on the Texture Properties of Cooked Long-Grain Rice

The effects of the degree of milling (based on surface lipids content [SLC]) on cooked rice physicochemical properties were investigated. Head rice yield (HRY), protein, and SLC decreased with increasing milling, while the percent of bran removed and whiteness increased. Results showed that SLC significantly (P < 0.05) affected milled as well as cooked rice properties across cultivar, moisture content (MC) at harvest, and location (Stuttgart, AR, and Essex, MO). Cooked rice firmness ranged from 90.12 to 111.26 N after milling to various degrees (SLC). The decrease in cooked rice firmness with increasing milling was attributed to the lowering of total proteins and SLC. Cooked rice water uptake increased with increasing degree of milling. Water uptake by the kernel during cooking dictated the cooked rice firmness. The increase in cooked rice stickiness with increasing degree of milling was attributed to an increase in starch leaching during cooking because of the greater starch granule swelling associated with a greater water uptake.