Role of secondary structures in the gelation of porcine myosin at different pH values

Secondary structures, gelation properties and their relationships in porcine myosin were studied by circular dichroism, dynamic rheological measurement and scanning electron microscopy. Gelling of porcine myosin involved a change in myosin conformation with protein-protein and protein-water interactions. The gelation properties were strongly pH and temperature dependent. Near the pI (pH 5.5 and 6.0), porcine myosin could spontaneously coagulate at 15°C resulting partially from the presence of more β-sheets. Myosin at pH6.5-9.0 began to form a gel at temperatures greater than 38°C. Heating caused α-helices to partially turn into β-sheets and random coils. Subsequently, myosin aggregated and formed a gel network. The gel strength decreased and the water-holding capacity (WHC) increased with increasing pH. Correlation analysis indicated that both the unfolding of α-helices and the formation of β-sheets favored the gelation of porcine myosin. A high β-sheet fraction prior to heating resulted in a low WHC of resultant gel. A compact and uniform gel was also obtained at pH6.5.     

Effect of sodium chloride and pH on the rennet coagulation and gel firmness

The effect of sodium chloride concentration and pH of milk at renneting on the rennet clotting time (RCT) and gel firmness was studied. The results showed that rennet clotting activity and gel firmness decreased with increasing NaCl concentration in milk. The RCT increased as the pH of the salted milk decreased below 6.4. Milk containing 5 and 10 g NaCl/100 g did not coagulate at pH 5.0. The gel firmness increased with decreasing milk pH to 6.2 for unsalted milk. However, the firmness decreased as the pH of milk decreased below 6.2. In case of using salted milk, the firmness decreased as the pH at renneting dropped below 6.4.     

Whey protein concentrate: Autolysis inhibition and effects on the gel properties of surimi prepared from tropical fish

Effects of whey protein concentrate (WPC) on autolysis inhibition and gel properties of surimi produced from bigeye snapper (Priacanthus tayenus), goatfish (Mulloidichthys vanicolensis), threadfin bream (Nemipterus bleekeri) and lizardfish (Saurida tumbil) were investigated. WPC (0–3%) showed inhibitory activity against autolysis in all surimi at both 60 and 65 °C in a concentration-dependent manner. Myosin heavy chain (MHC) of surimi was more retained in the presence of WPC. Breaking force and deformation of kamaboko gels of all surimi increased as added levels of WPC increased (P < 0.05). This was associated with lower levels of protein degradation, as evidenced by the decrease in trichloroacetic acid-soluble peptide content (P < 0.05). WPC at 3% (w/w) significantly decreased the whiteness of gels. However, water-holding capacity of kamaboko gels was improved with increasing concentration of WPC. The microstructure of surimi gels generally became denser with the addition of WPC.     

Effect of corn starch on the structure,physicochemical and gel properties of hairtail (Trichiurus haumela) myosin

Effects of corn starch on structure, physicochemical and gel properties of myosin were investigated. Particle size, turbidity, surface hydrophobicity and total sulphhydryl groups of myosin were significantly increased with the increasing of starch concentration, and the increase range of high amylose corn starch (HACS) group was the most significant. Tryptophan residue of myosin was buried, and fluorescence quenching occurred after starch was added. HACS made myosin secondary structure was changed from α-helix to β-sheet. Confocal laser scanning microscopy (CLSM) photographs showed starch was encapsulated by aggregated myosin, which made the protein binding tighter and more beneficial to the formation of gel. Moreover, strength and water holding capacity of heat-induced myosin gels containing starch were significantly higher than that of pure myosin. Starch, especially HACS, could induce myosin aggregation before heating gelatinisation and promote myosin self-assembly at low temperature, which may be conducive to forming a higher quality gel after heating.     

三种添加物对鱼肉猪肉复合凝胶品质的影响

研究了三种外源添加物(大豆蛋白、液体蛋清、转谷氨酰胺酶)对鱼肉猪肉复合凝胶品质的影响。结果表明,添加大豆蛋白和液体蛋清均可提高复合凝胶的破断强度和凝胶强度;添加液体蛋清还可提高复合凝胶的明度L*和白度W,而添加大豆蛋白则会降低明度L*和白度W。添加转谷氨酰胺酶可显著提高复合凝胶的凝胶强度,且鱼肉加盐斩拌后加入转谷氨酰胺酶、再斩拌,然后加入猪肉或者鱼肉、猪肉加盐混合斩拌后再加入转谷氨酰胺酶,两种效果较好。      

超声波处理对棉纤维结构与性能的影响

用XRD、TGA、SEM等方法测定并分析了超声波处理对棉纤维的形态结构、结晶形态、热性能和力学性能的影响,力求为超声波在棉织物染整加工中的应用奠定理论基础。实验结果表明:超声波处理改变了棉纤维的形态结构,使棉纤维的天然扭曲消失,纤维表面变得粗糙;使棉纤维的结晶度降低,结晶尺寸减小;棉纤维的热裂解温度降低,热稳定性变差;棉织物的断裂强度降低,断裂延伸度提高。     

干热处理和pH偏移对汉麻分离蛋白/卡拉胶接枝物凝胶性质的影响

摘 要： 目的：利用植物蛋白代替动物蛋白有利于减少动物蛋白消耗，改善汉麻分离蛋白在蛋白凝胶中的应用前景。方法：以未经pH偏移及干热处理组分为空白对照，研究了不同干热处理时间（0、24、48、72 h ）和pH偏移（2、12）条件对汉麻分离蛋白/卡拉胶接枝物凝胶硬度、流变特性、水分分布和微观结构的影响。结果：与对照组相比，经干热处理和pH偏移处理的蛋白凝胶硬度、持水性和储能模量（G’ ）均显著提高（P<0.05），其中干热处理48 h和pH 2偏移处理组分是对照组的1.38、1.10和19.45倍。干热处理和pH偏移增加了凝胶中不可流动水的含量（92.88%）且电镜结果显示凝胶的微观结构逐渐致密，pH2偏移和干热处理48h最光滑，复合凝胶中蛋白的α-螺旋的含量减少，β-折叠含量增加（P<0.05），这表明蛋白分子内部的有序结构增加。结论：综上，在pH 2偏移干热处理48h条件下，汉麻分离蛋白凝胶性质最佳研究表明，这为汉麻分离蛋白在食品工业中的应用提供了新的理论依据。     

原料乳预酸化的pH对Mozzarella干酪功能特性的影响

采用直接酸化工艺,用柠檬酸将原料乳pH分别调节为6.37、6.09和5.73,研究了原料乳不同酸化的pH对Mozzarella干酪功能特性的影响。结果表明,随着原料乳预酸化pH的降低,干酪的TPA硬度显著降低(p<0.05),TPA粘性显著增加(p<0.05),对TPA弹性影响不显著,干酪的融化性和拉伸性显著增强(p<0.05)。      

Effect of cassava starch gel,fish gel and mixed gels and thermal treatment on structure development and various quality parameters in microwave vacuum-dried gel slices

Three types of gels including tapioca starch gel, fish gel and mixed gels of both (cassava starch: fish = 1:1) were prepared under different thermal conditions. Slices (4 mm thick) derived from these the gels were dehydrated to a final moisture content of 7% in a microwave vacuum dryer. The micro/macro structure and selected quality parameters of the dried gel chips were measured e.g. shape, relative volume, bulk density, texture, color, sensory and microstructure. The measured gel characteristics were analyzed and related to structure development and selected quality attributes. Both fish and mixed gel chips expanded in both diameter and thickness while the starch gel chips shrank in diameter, but expanded in thickness; the former two represented a continuous cross-section composed of a cellular structure and expanded uniformly in thickness while the latter developed an open cross-section and the center expanded more than the edge region, indicative of a disc and pillow shape, respectively. The higher severity of thermal treatment favored greater expansion and reduced hardness and bulk density, and increased crispness in the mixed gel chips; they developed a smooth and fine surface and pore wall but decreased lightness and whiteness due to starch gelatinization in the starch and mixed gel chips. Only the partial dried mixed gel chips are acceptable to the panelists. A homogenous co-gel in the mixed gel contributed to the higher uniform expansion and cellular structure. These findings will help developing the restructured microwave vacuum-dried product by blending the starch with the fish.     

NaCl浓度和pH对肉糜中脂肪微粒蛋白吸收量及凝胶特性的影响

研究了不同浓度(0.2mol/L和0.6mol/L)NaCl和pH(6.0和6.5)对肉糜中脂肪微粒表面蛋白吸收量、乳化稳定性、凝胶硬度和微观结构的影响。研究结果表明,高浓度(0.6mol/L)NaCl能够显著提高肉糜中脂肪微粒表面蛋白吸收量和单位界面膜蛋白吸收量,有效地改善肉糜乳化稳定性、凝胶硬度和微观结构等指标(P<0.05)。但在相同浓度(0.2mol/L和0.6mol/L)NaCl条件下,较大pH(6.0和6.5)对脂肪微粒表面蛋白吸收量、单位界面膜蛋白吸收量、乳化稳定性、凝胶硬度和微观结构等指标无作用效果(P>0.05)。因此,本实验认为0.6mol/LNaCl(pH6.0和6.5)的提取条件下能够充分剪切肌肉蛋白和脂肪,肉糜乳化性能好。      

Analysis of fish and squid myofibrillar proteins by capillary sodium dodecyl sulfate gel electrophoresis: actin and myosin quantification

 A capillary electrophoretic method for the separation and quantification of fish and squid myofibrillar proteins was developed. The method uses sodium dodecyl sulfate and β-mercaptoethanol for solubilization and analysis of myofibrillar protein subunits. The separation of the different polypeptides is achieved by the sieving effect of the gel inside the capillary. A calibration curve for myosin heavy chain and actin UV absorbance quantification of these proteins was developed. Received: 2 November 1999 / Revised version: 16 February 2000     

微波预处理对热诱导小麦面筋蛋白凝胶性质和微观结构的影响

研究微波预处理对热诱导小麦面筋蛋白凝胶性质和微观结构的影响。结果表明,当微波条件为700 W功率,处理时间90 s时,热诱导小麦面筋蛋白凝胶强度和持水性达到最大值158.6 g/cm~2和83.13%,比对照组分别提高83.4%和162%,此时凝胶中非冻结水的比例也显著提高。随着微波功率从70 W增加到700W,微波处理时间从10 s增加到90 s,热诱导小麦面筋蛋白凝胶的表面疏水性逐渐降低。微波预处理还显著增大了热诱导小麦面筋蛋白凝胶体系中的疏水相互作用,疏水相互作用的贡献从0.85 g/L增大到1.69 g/L。微波预处理的热诱导小麦面筋蛋白凝胶具有更为致密、均匀的微观三维网络结构。     

Effect of malondialdehyde oxidation on structure and physicochemical properties of amandin

Oil, accounting for 45% of almonds, is easily oxidised and can further induce the protein oxidation to reduce their quality. Structure and physicochemical properties of amandin, the main water-soluble protein in almonds, inducing oxidation by malondialdehyde (MDA) were investigated. The results showed that the content of carbonyl group increased from 5.23 to 33.25 nmol mg⁻¹ of protein with the increase in MDA concentration (P < 0.05). However, the sulphydryl content, surface hydrophobicity, particle size and the absolute value of ζ-potential first increased and then decreased. Fourier-transformed infrared spectroscopy (FT-IR) confirmed that the structure of amandin changed from order to disorder. Fluorescence spectroscopic analysis revealed that mild oxidation (0–0.1 mmol L⁻¹ MDA) exposed hydrophobic groups of the protein. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) suggested that protein oxidation promoted crosslinking between protein molecules. Furthermore, protein oxidation markedly declined the total amino acid content of amandin (P < 0.05). In conclusion, MDA oxidation changed the structure and amino acid content of amandin, and caused the protein aggregate and crosslink through hydrophobic interaction and electrostatic interaction.     

Naturally occurring variations in milk pH and ionic calcium and their effects on some properties and processing characteristics of milk

Considerable variations were found for ionic calcium and pH in milk from individual cows. Milk with lower pH tended to have a higher Ca²⁺ concentration, although the relationship was weak. Milk samples with a higher Ca²⁺ concentration and lower pH produced less sediment during in‐container sterilisation, which was contrary to expectations. Rennet coagulation time was lower for milk with a higher Ca²⁺ concentration, but curd firmness was not related to Ca²⁺ concentration. There was a poor correlation between the pH reduction caused by acid addition and that resulting from increasing temperature. Sediment formation was related to pH change at high temperature.     

Characterization of fish gelatin from surimi processing wastes: Thermal analysis and effect of transglutaminase on gel properties

Fish gelatin extraction from wastes of fish Herring species (Tenualosa ilisha) was carried out by a series of pretreatment with 0.2 M Ca(OH)₂ followed by 0.1 M citric acid and final water extraction at 50 °C for 3 h. The resulting fish gelatin preparation was evaluated for its dynamic viscoelastic properties, gelling and melting temperatures and gel strength. The gelling and melting temperatures of gelatin samples (at 6.67%, w/v) were obtained from differential scanning calorimetry and rheological studies. The melting temperature of extracted fish gelatin (EFG) obtained ranged from 16.2 to 16.7 °C compared to that of commercial fish gelatin gel (CFG), from 23.7 to 25.6 °C and halal bovine gelatin (HBG), from 26.5 to 28.7 °C. On the other hand, gelling temperatures of EFG, CFG and HBG ranged from 5.1 to 5.2 °C, 11.9 to 17.46 °C, and 12.6 to 19.33 °C, respectively. EFG gave gels with a considerably lower G′ values than CFG and HBG. The bloom strength of EFG gels at 6.67% (w/v) was 69.03 g which was much lower than HBG (336.2 g) and CFG (435.9 g). Enzyme transglutaminase was added in the amounts of 0.5, 1.0, 3.0 and 5.0 mg/g gelatin to modify the gel properties of the extracted fish gelatin. The modified EFG gels obtained had higher gel strengths of 101.1 g and 90.56 g with added transglutaminase of 1.0 and 3.0 mg/g, respectively. However with addition of 5.0 mg/g enzyme the gel strength increased only up to 75.06 g. SDS-PAGE of extracted gelatin gel showed protein band intensities for α1-chains and 53 kDa but in gels added with higher concentration of transglutaminase, these protein band intensities seemed to disappear.     

Post-mortem degradation of myosin heavy chain in intact fish muscle: Effects of pH and enzyme inhibitors

Fish muscle is rapidly degraded during post-mortem storage, due to proteolytic enzymes acting probably both on muscle cells and connective tissue. In this work we have developed a model system which may be used to study the enzymatic degradation occurring in intact post-mortem fish muscle. Degradation of myosin heavy chain (MHC) was monitored in muscle with pH adjusted to 6.05, 6.3 and 6.9 and in the presence of the enzyme inhibitors PMSF, EDTA, phenanthroline, pepstatin A, antipain, E-64 and the cysteine proteinase activator dithiothreithol (DTT). After storage, myofibrillar proteins were isolated and MHC-specific antibodies used to study the degradation in the different samples. MHC from muscle with pH 6.05 and 6.3 was degraded, while no severe degradation was observed at pH 6.9. Introduction of enzyme inhibitors into the muscle tissue clearly showed that mainly cysteine and aspartic proteinases are responsible for the in situ MHC degradation. This is supported by the severe breakdown of MHC in the muscle samples containing DTT.     

Contribution of protein conformation and intermolecular bonds to fish and pork gelation properties

Changes in protein conformation and intermolecular interactions during gelling were measured to explore the basis for the different gel properties of fish and pork pastes after heat treatment. Raman spectral analysis revealed that, in the unheated pork sample, more tyrosine residues were buried or involved as hydrogen bond donors than in the fish counterpart. Hydrophobic interaction increased and then decreased with increasing temperature, reaching a maximum value at 60 and 70 °C for fish and pork samples, respectively. Formation of disulfide bonds mainly occurred at 70–80 °C for fish, and 50–60 °C for pork samples. Incubation at 4–40 °C induced abundant non-disulfide covalent cross-linking in fish, but not in pork samples. The differences in protein conformation and intermolecular bonds possibly contributed to diverse gelation properties between fish and pork samples. At 40 and 50 °C, the breaking force of fish samples was significantly higher than that of pork. However, higher breaking force was observed in pork samples above 60 °C. Pre-incubation at 40 °C significantly increased the breaking force of the cooked fish sample, but had little effect on the breaking force of the cooked pork sample. In addition, incubating pork paste at 40 °C could decrease the cooking loss of the cooked pork sample.     

Primary and secondary structure of novel ACE-inhibitory peptides from egg white protein

The primary structure of novel angiotensin converting enzyme (ACE) inhibitory peptide from egg white protein was investigated, and secondary structure of the peptide was explored for the first time. The potential effects of bioactive peptides were submitted to bioactivity screening with ACE inhibitory activity, antioxidant property, and anticoagulation activity. Bioactive peptides from egg white protein were characterized by LC tandem mass spectrometric, and secondary structures of those peptides were investigated by FT-IR. Our results showed that total 11 bioactive peptides with three new and eight known structures were identified with LC/MS/MS, which then were synthesized by Fmoc solid phase method. Peptide Thr-Asn-Gly-Ile-Ile-Arg (TNGIIR) exhibited higher activity against ACE to other two new peptides. The concentration of the peptide TNGIIR, necessary to inhibit 50% the activity of ACE was 70 μM. Results also suggested that the secondary structural differences between peptides could also influence the ACE inhibition capacity. Thus, it appears that primary and secondary structure of peptide plays the potential role inhibiting the ACE activity.     

Effect of furcellaran incorporation on gel properties of sardine surimi

Effects of furcellaran, powder (FURP) and solution (FURS), incorporation (2-8% based on surimi solid content) on gel properties of sardine (Sardinella albella) surimi were investigated. Breaking force, hardness, gumminess, chewiness, and whiteness of gels increased with increasing FURP or FURS contents (P < 0.05), where the highest values were observed in gel containing 8% FURS (P < 0.05). Expressible moisture content of gel decreased as levels of furcellaran increased (P < 0.05). The gel deformation, springiness, and cohesiveness were not affected by furcellaran (P > 0.05) as well as the polymerization of myosin heavy chain as visualised by SDS-PAGE. FURP and FURS could enhance the interconnection between chains during heating as indicated by the higher G′. Addition of furcellaran, especially FURS, up to 6% increased sensory property of resulting gel (P < 0.05). Anywise, an abatement in overall likeness score was found in gel with 8% FURS. Finer and denser microstructural network was observed in gel containing 6% FURS when compared to control. Therefore, furcellaran, prepared as solution, at an appropriate level can improve the overall quality of sardine surimi.