脂肪氧合酶催化亚油酸氧化对花生蛋白结构的影响

利用脂肪氧合酶催化亚油酸氧化的反应体系对花生分离蛋白进行不同程度的氧化修饰,通过研究不同亚油酸含量下花生分离蛋白的羰基值、游离巯基含量、粒径分布、表面疏水性、溶解度以及荧光光谱的变化规律,从而探讨氧化对花生分离蛋白结构的影响。结果表明:随着反应体系中底物亚油酸含量的增加,花生分离蛋白的羰基值先增加后略微下降,游离巯基含量下降,表面疏水性先增加后减小,说明氧化后花生分离蛋白的结构已经发生了改变。其粒径分布和溶解度的变化规律可以表征花生分离蛋白氧化聚集体的状态,同时其荧光峰位λmax的变化规律可反映花生分离蛋白三级结构的具体变化,表明脂肪氧合酶催化亚油酸氧化可以诱导花生分离蛋白分子发生聚集,使其结构发生显著变化。     

脂肪氧合酶催化亚油酸氧化对稻米醇溶蛋白的影响

研究脂肪氧合酶催化亚油酸氧化模拟体系中,不同质量浓度的底物亚油酸对稻米醇溶蛋白羰基值、总巯基及活性巯基、表面疏水性及蛋白质的二级结构的影响。试验结果表明:氧化后,稻米醇溶蛋白的活性巯基呈现先下降后上升的趋势,总巯基呈现上升趋势。羰基值及表面疏水性均呈现一定的上升趋势,圆二色谱测定结果表明蛋白质的二级结构也发现了相应的变化。综上所述,脂肪氧合酶催化亚油酸氧化可以诱导稻米蛋白结构发生规律性变化。     

氢过氧化物氧化对核桃蛋白结构和乳化特性的影响

采用脂肪氧合酶催化亚油酸氧化的反应体系进行氧化修饰核桃分离蛋白,代表脂质氢过氧化物氧化核桃分离蛋白。通过对不同亚油酸含量下核桃分离蛋白的结构指标进行表征,探讨氢过氧化物氧化对其结构的影响。采用氧化修饰后的核桃分离蛋白制备乳状液,对其乳化特性进行综合评价,结果表明,随着亚油酸添加量的增加,羰基和二硫键含量增加,巯基含量减少,表面疏水性下降,溶液可溶性聚集体转变成不可溶性聚集体,粒径变大。乳液内源荧光光谱分析检测到核桃分离蛋白乳液的构象发生改变。不同的氧化程度,乳液油滴粒径分布不同。适度的氧化,使蛋白质结构展开,暴露出更多的疏水基团,形成可溶性聚合物,乳液油滴粒径降低,乳化活性有所提高,进一步氧化,促进液滴的聚合,致乳化活性下降。结构和乳化功能指标之间的皮尔逊相关系数表明,由HPODE氧化引起的蛋白质结构的改变对核桃蛋白乳化性的影响并不显著。     

加工方式对牛肉蛋白质氧化的影响

为了研究热加工过程中蛋白质氧化的变化规律,以牛腱子和牛肩为研究对象,对其进行不同温度（40、50、60、70、80、90、100 ℃,保温时间30 min）以及70 kPa高压下保温不同时间（15、20、25、30、35 min）的热处理,通过分析羰基含量、巯基含量、蛋白粒径、表面疏水性、SDS-PAGE电泳以及蛋白二级结构等的变化,结果表明:温度可以导致蛋白羰基含量增加,巯基含量先升高后降低,蛋白粒径增大,蛋白表面疏水性先增大后减小;高压导致蛋白羰基含量增加,巯基含量减少,蛋白粒径先增大后减少,蛋白疏水性逐渐增加。另外,不同热处理下肌原纤维蛋白发生了明显的降解聚集,出现大量小分子蛋白质。红外研究发现在热加工过程中,肌原纤维蛋白的二级结构不断转变,促使α-螺旋、β-折叠和无规则卷曲之间的转换。可知,温度和压力都能促进牛肉蛋白的氧化并能够改变牛肉蛋白的化学作用力及二级结构。本研究为低温牛肉产品的工业化调控提供理论依据。     

脂质氢过氧化物氧化对核桃分离蛋白结构的影响

采用脂肪氧合酶催化亚油酸构建脂质氢过氧化物——核桃分离蛋白氧化体系,研究脂质氧化反应中产生的氢过氧化物对核桃分离蛋白结构特性的影响。结果显示,向100 m L 0. 05 mg/m L蛋白溶液中添加9 m L亚油酸,核桃分离蛋白溶解度下降至6. 89%,游离巯基含量由2. 82μmol SH/g下降至1. 92μmol SH/g,羰基含量由2. 26 nmol/mg增加至4. 23 nmol/mg,表明氧化后核桃蛋白理化特性产生变化。圆二色谱分析表明,蛋白质氧化导致核桃分离蛋白α-螺旋和β-折叠含量下降,β-转角和无规则卷曲含量上升。随着亚油酸添加量的增加,蛋白质氧化程度加大,核桃分离蛋白表面疏水性从429. 66下降到405. 24。内源荧光最大荧光峰位红移,内源荧光强度下降,此结果表明,脂肪氧合酶(lipoxygenase,LOX)催化亚油酸氧化诱导核桃分离蛋白聚集,并进一步使蛋白质二级和三级结构发生变化。体积凝胶色谱法结果显示,核桃分离蛋白氧化后,小分子多肽含量下降,高分子量聚集体含量增加,说明脂质氢过氧化物氧化修饰导致核桃蛋白结构改变,形成氧化聚集体。     

羟自由基诱导蛋白质氧化损伤的研究

作者利用Fe2+/H2O2/Asc氧化体系制备羟自由基,通过分光光度法、荧光分光光度法及FT-IR,研究不同浓度过氧化氢(H2O2)对牛血清蛋白(BSA)巯基及活性巯基、表面疏水性、羰基含量及蛋白质的二级结构的影响。结果表明:总巯基及活性巯基的含量都有着不同程度的减少,表面疏水性和羰基含量均随着氧化剂浓度的增加和氧化时间的延长而增加,蛋白质的二级结构也发生了改变。     

蛋白质氧化对核桃蛋白质结构的影响

为了深入明晰氧化对核桃蛋白质结构性的影响,本文以核桃分离蛋白为研究对象,采用不同浓度的2,2'-盐酸脒基丙烷(AAPH)热降解形成烷过氧自由基(ROO·)代表脂质过氧化反应过程中产生的脂质自由基,对核桃分离蛋白进行氧化修饰得到不同氧化程度的核桃氧化蛋白。对不同氧化程度蛋白的羰基、巯基和总巯基,表面疏水性(H₀)、内源荧光、粒径分布、相对分子质量和二级结构进行研究和分析。结果表明,随着AAPH浓度的增加,核桃氧化蛋白羰基含量显著增加(p<0.05),巯基和H₀显著降低(p<0.05);圆二色谱结果表明,随着AAPH浓度的增加,核桃蛋白α-螺旋结构和β-折叠含量下降,而无规卷曲结构含量增加,说明氧化破坏了蛋白质的二级结构,使有序结构变为无序结构。最大内源荧光值降低,且出现蓝移现象,说明烷过氧自由基氧化导致分子聚集,使得色氨酸残基包埋;粒径分布结果表明,核桃氧化蛋白可溶性聚集体可被共价交联和非共价聚集转变成不可溶性部分;体积排阻色谱结果表明,核桃氧化蛋白相对分子质量随着氧化浓度的增大聚集程度逐渐增大。研究结果表明,核桃蛋白在烷过氧自由基(ROO·)氧化体系中产生了显著氧化作用并导致其结构发生一定的改变,为进一步阐明核桃蛋白氧化机理提供理论依据。     

蛋白质氧化对核桃蛋白界面性质的影响

采用不同浓度(0～10 mmol/L)的丙二醛替代脂质过氧化反应中产生的活性醛氧化核桃蛋白,研究蛋白质氧化对核桃蛋白界面性质的影响。试验结果表明:随着丙二醛浓度的增加,核桃蛋白发生显著氧化,羰基含量从3.12 nmol/mg增至20.61 nmol/mg。随着核桃蛋白氧化程度的增加,核桃蛋白粒径分布发生偏移,蛋白质相对分子质量先减小后增加,表面张力呈先下降后上升的趋势,Zeta电位绝对值、表面疏水性、起泡性和泡沫稳定性呈先增加后减小的趋势。当蛋白质氧化体系中的丙二醛浓度为0.1 mmol/L时,表面张力达到最小值,即55.64 mN/m;核桃蛋白的Zeta电位绝对值、表面疏水性、起泡性和起泡稳定性达到最大值,分别为10.38 mV,476,21.43%,17.80%。研究结果表明:丙二醛氧化诱导使得核桃蛋白的分子结构和界面性质发生改变,较低程度的氧化有利于改善核桃蛋白的界面性质。     

丙二醛氧化修饰对核桃分离蛋白乳化性质的影响

以不同浓度(0~10 mmol/L)的丙二醛氧化处理的核桃分离蛋白为研究对象,探究脂肪氧合酶(LOX)诱导多不饱和脂肪酸发生脂质过氧化反应过程中产生的活性醛类对核桃分离蛋白乳化性质的影响。研究结果表明:随着丙二醛浓度从0增至10 mmol/L,核桃分离蛋白发生显著氧化,羰基值从3.12 nmol/mg增至20.61 nmol/mg;表面疏水性呈现先上升后下降的趋势;乳液内源荧光强从419.97降至109.86,最大吸收波长λmax发生蓝移;乳液粒径和乳析率均呈现先减小后增大的趋势;乳液Zeta电位值和乳化稳定性则呈现先增加后减小的趋势;而乳化活性从46.57 m^2/g降至26.26 m^2/g。当丙二醛浓度为0.1 mmol/L时,核桃分离蛋白乳液粒径和乳析率达到最小值,Zeta电位值和乳化稳定性达到最大值,说明适度的丙二醛氧化使得核桃分离蛋白的乳液更加稳定,而氧化对蛋白质乳化活性产生负面影响。     

3种多酚干预过氧自由基诱导核桃蛋白氧化的研究

为研究没食子酸、儿茶素、杨梅素对核桃蛋白氧化稳定性的影响,以2,2-偶氮二异丁基脒二盐酸盐(2,2’-Azobis[2-Methylpropionamidine]dihydrochloride,AAPH)为引发剂,建立过氧自由基氧化核桃蛋白的模拟体系,分析经3种多酚干预后其羰基、巯基、内源荧光、表面疏水性、溶解度等指标的变化规律。结果表明,经过氧自由基氧化后,核桃蛋白的羰基、巯基含量和表面疏水性显著增大,荧光强度和溶解度显著下降(P<0.05)。而经3种多酚干预后,核桃蛋白的羰基含量和表面疏水性相比氧化组显著降低,最大降幅分别为72.2%和58.0%;巯基含量和溶解度相比氧化模型组显著增加,最大增幅分别为2.1倍和53.0%;此外,核桃蛋白的荧光强度显著上升(P<0.05)。相关性热图和主成分分析进一步说明多酚干预可在一定程度降低核桃蛋白的氧化程度,且3种多酚干预对核桃蛋白的氧化指标存在显著区别。总体上,儿茶素对过氧自由基氧化核桃蛋白的抑制效果最好,可为核桃制品的合理储藏和加工提供一定参考。     

自由基氧化引起乳清蛋白理化性质变化的研究

研究了在羟基自由基氧化体系中,不同H2O2浓度(1～20 mmol/L)及不同FeCl3浓度(0.1～2 mmol/L)对乳清蛋白羰基、巯基、二聚酪氨酸等理化性质的影响.每种氧化条件的氧化时间分别为1,3,5 h..结果表明:氧化显著地影响了乳清蛋白的理化性质,同未氧化的对照组乳清蛋白相比,经过5 h氧化,所有浓度的FeCl3体系中,羰基增加3倍以上;所有条件下的巯基损失均达40%以上;在浓度为20 mmol/L的H2O2或2 mmol/L的FeCl3中,二聚酪氨酸分别增加了5倍和7倍.并且发现,在不同的FeCl3条件下其变化趋势更为迅速.由此可知,氧化极大程度地改变了蛋白的理化性质,并可能导致蛋白结构的改变,进而可能影响其功能性质.     

Effects of malondialdehyde-induced protein oxidation on the structural characteristics of rice protein

Malondialdehyde (MDA) was selected to represent a secondary by-product of lipid peroxidation during rice ageing. This study aimed to investigate the effects of MDA modification on the structural characteristics of rice protein. The results showed that as MDA concentration increased, rice protein carbonyl and disulphide groups increased, but sulphydryl content decreased. The blue shift of maximum fluorescence peak, the decrease of rice protein intrinsic fluorescence intensity and the reduction of surface hydrophobicity indicated the formation of protein aggregates caused by MDA oxidative modification. The results of molecular weight distribution and particle size distribution showed that MDA modification resulted in the formation of soluble protein aggregates, and the decrease of rice protein solubility indicated that insoluble protein aggregates were formed. Results of protein electrophoresis showed that MDA modification contributed to rice protein aggregation via non-disulphide covalent bonds. The results showed that rice protein gradually aggregated with increasing MDA concentration.     

Relationship between N-nitrosodiethylamine formation and protein oxidation in pork protein extracts

The relationship between protein oxidation and N-nitrosodiethylamine (NDEA) formation was assessed. Packaged fresh minced pork was stored at 2 ± 1 °C for 0, 4, 7, and 10 days. Myofibrillar protein (MP), sarcoplasmic protein (SP), and muscle homogenate were obtained from the stored minced pork and allowed to react with sodium nitrite at 80 °C for 1 h. The relationship between NDEA formation and indexes of protein oxidation was assessed by principal component analysis. The formation of NDEA in meat was correlated with protein and lipid oxidation. Lipid oxidation promoted the formation of NDEA and was positively correlated with protein oxidation. Carbonyl compounds from oxidized MP and free amines from dissociated SP promoted NDEA formation. Therefore, the effect of protein oxidation on NDEA formation depends on the type of protein and its oxidation products.     

稻谷蛋白质氧化及其对稻谷陈化品质的影响

综述了稻谷陈化过程中一些主要化学成分如淀粉、脂肪、蛋白质等的变化,以及直链淀粉含量、黏度、脂肪酸含量等稻谷陈化品质评价指标,创新性地从蛋白质氧化的角度探讨稻谷蛋白质氧化对陈化品质的影响,主要包括蛋白质单独作用、蛋白质与淀粉相互作用及蛋白质、淀粉及脂肪共同作用改变稻谷陈化品质,指出目前稻谷蛋白质氧化及陈化品质变化仍存在的一些问题,为进一步揭示稻谷陈化机理和提出更有效的稻谷陈化品质评价指标提供理论依据。     

蛋白质氧化机制及其评价技术研究进展

肉制品在贮藏一段时间后,颜色会变成让人难以接受的程度,为肉制品的销售带来不利影响。肉制品颜色的变化在一定程度上与蛋白质氧化有很大地关系。对肉制品中蛋白质氧化的研究有助于延缓肉制品颜色的劣变,提高肉制品的货架期。主要就蛋白质氧化的机理、氧化后蛋白质发生的主要变化以及氧化后蛋白质的评价技术作了阐述,旨在建立食品中蛋白质氧化的完善评价机制。     

Higher drip loss is associated with protein oxidation

The study aimed to investigate the mechanisms underlying drip loss in meat other than those related to post mortem energy metabolism. The study was carried out on M. longissimus thoracis assessing carcass and meat quality traits plus metabolic parameters and drip loss. Based on the data obtained, three drip loss groups were established: low, medium and high. Heat treatments were performed at 100 °C for 30 min. Physicochemical protein modifications were assessed before and after cooking. IMF and L* were higher in the high drip loss group, whereas b* was higher in the medium drip loss group. Residual glycogen, glucose and glycolytic potential were higher in LT muscle from the high drip loss group. Before cooking, protein surface hydrophobicity and carbonyl levels were similar in the three groups. However, after cooking, carbonyl, oxidized actin and oxidized aggregates were higher in the high drip loss group, suggesting that protein oxidation may affect the water holding capacity of meat.     

Lipid and protein oxidation of chicken breast rolls as affected by dietary oxidation levels and packaging

The objective of this study was to determine the effects of dietary treatment and packaging on the oxidative stability of breast rolls. A total of 120 4-wk-old broiler chickens were randomly assigned to control, oxidized diet (5% oxidized oil, PV = 100), or antioxidants-added diet (500 IU vitamin E + 200 ppm BHA) and fed for 2 wk. Breast muscles were separated from the carcasses and breast rolls were prepared. The rolls were cooked in a smoke house (85 °C) to an internal temperature of 74 °C, cooled, sliced to 2-cm thick pieces, individually packaged in oxygen permeable bags or vacuum-packaged in oxygen impermeable bags, and stored in a 4 °C cold room for 7 d. Lipid, protein oxidation and volatiles were determined at 1, 4, and 7 d of storage. Dietary supplementation of antioxidants significantly reduced lipid oxidation (TBARS) and protein oxidation (carbonyls) in breast rolls, and the effect of dietary antioxidants on lipid oxidation was more pronounced than protein oxidation. Chicken breast rolls from antioxidants treatment group produced significantly lower amounts of hexanal and pentanal than those from control and oxidized oil treatments (P < 0.05). However, dietary oxidized oil did not increase lipid and protein oxidation in breast rolls. Vacuum-packaging significantly delayed the onset of lipid oxidation and protein oxidation in chicken rolls during 7-day refrigerated storage (P < 0.05). Therefore, it is suggested that appropriate use of dietary supplementation of antioxidants in combination with packaging could minimize lipid oxidation in chicken breast rolls.     

肉制品中脂肪氧化与蛋白质氧化及相互影响

近年来,食品中脂肪氧化与蛋白质氧化之间的联系在国内外引起了广泛关注。肉制品中脂肪及蛋白质的氧化能产生特征风味,但更多的是引起风味的劣化和颜色的褪变,且脂肪氧化和蛋白氧化之间是相互关联的,彼此相互促进。针对由脂肪氧化和蛋白质氧化影响肉制品品质的问题,就脂肪氧化与蛋白质氧化机理、肉制品中脂肪与蛋白质氧化的相互影响关系及控制措施进行综述。     

Effect of heat treatment on protein oxidation in pig meat

We investigated the oxidative mechanisms and identified the target protein induced by heat treatment. The study was carried out on M. longissimus thoracis from Galia and Redone pigs. Post mortem metabolic parameters and drip loss were determined. Heat treatment was performed at 100 °C for 10 and 30 min. Physicochemical state of the protein, TBA-RS and Schiff bases were assessed. Protein aggregates were evaluated and the protein target of oxidation studied. Muscles from Galia had higher residual glycogen and drip loss. Heat treatment increased surface hydrophobicity, carbonyl, protein aggregate and Schiff bases and TBA-RS whatever the treatment time. Immunoblotting revealed oxidized myosin, oxidized actin and high molecular weight proteins after 30 min cooking. Oxidation products were significantly correlated with drip loss, suggesting a possible reduced ability of oxidized proteins to retain water. Moreover, residual glycogen was positively correlated with oxidized myosin, suggesting a possible role of glycogen as a glucose donor.