4 种香辛料提取物对猪肉肌原纤维蛋白抗氧化特性的影响

以猪肉肌原纤维蛋白为研究对象,采用羟自由基氧化体系对蛋白进行0、1、3、5?h的氧化,分析4?种香辛料提取物对肌原纤维蛋白抗氧化特性的影响。结果表明：丁香、迷迭香和桂皮提取物的添加显著抑制蛋白羰基的形成（P＜0.05）,甘草提取物的作用不显著;丁香和迷迭香提取物对Ca-ATP酶活性下降具有显著抑制作用（P＜0.05）,桂皮和甘草提取物无积极作用;4?种提取物均具有较强的抑制表面疏水性增加的作用;丁香、迷迭香和甘草提取物对蛋白溶解性下降具有很好的抑制作用,桂皮提取物无积极作用;4?种提取物的添加均不同程度地加速了蛋白巯基的下降。综上,丁香和迷迭香提取物具有较好的抑制猪肉肌原纤维蛋白氧化的作用。     

羟自由基氧化对高白鲑肌原纤维蛋白降解的影响

为了探究羟自由基氧化对鱼肉宰后贮藏期间肌肉蛋白降解的影响,以高白鲑为研究对象,取其背部肌肉进行氧化,研究羟自由基氧化对高白鲑肌原纤维蛋白降解的影响。设置4 组不同浓度的氧化体系（1 mmol/L H2O2＋0.2 mmol/L FeCl3、5 mmol/L H2O2＋0.4 mmol/L FeCl3、10 mmol/L H2O2＋0.8 mmol/L FeCl3、20 mmol/L H2O2＋1.0 mmol/L FeCl3）,并分别室温氧化0、15、30、60、90 min,以羰基含量确定最适氧化浓度和时间;氧化样品于4 ℃贮藏,分别在0、1、7、14 d取样,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质免疫印迹检测肌球蛋白重链（myosin heavy chain,MHC）、肌动蛋白、肌间线蛋白和肌钙蛋白T等蛋白的降解程度。结果表明：5 mmol/L H2O2＋0.4 mmol/L FeCl3的氧化浓度下孵育60 min为本实验最适氧化条件;氧化组MHC的降解率比未氧化组高,羟自由基氧化显著促进了MHC的降解（P＜0.05）;肌动蛋白变化不显著（P＞0.05）,自然降解占主导地位;氧化组中肌间线蛋白和肌钙蛋白T的降解率低于未氧化组,羟自由基氧化抑制了肌间线蛋白和肌钙蛋白T的降解。综上所述,羟自由基氧化主要促进肌纤维中粗丝蛋白的降解,对细丝蛋白的降解没有明显影响,对细胞骨架蛋白和连接蛋白的降解起到抑制作用。     

大豆分离蛋白对肌原纤维蛋白加热过程中结构及流变特性的影响

大豆分离蛋白（soybean protein isolate,SPI）作为优质的植物蛋白常被用于肉制品加工中,以提高产品产量和质地。研究添加SPI对肌原纤维蛋白（myofibrillar protein,MP）凝胶特性及MP加热过程中结构和流变特性的影响。结果表明：添加10%、20% SPI能提升混合凝胶的凝胶强度及保水性（P＜0.05）;加热过程中混合蛋白凝胶二级结构发生改变,但其变化规律尚不明确;添加SPI使混合凝胶储能模量及损耗模量下降;混合凝胶上清液十二烷基硫酸钠-聚丙烯酰胺凝胶电泳图谱显示,肌球蛋白重链、肌动蛋白、SPI部分亚基均是参与凝胶形成的蛋白质。     

壳聚糖复合保鲜对冷藏草鱼肌肉蛋白质组的影响

为深入研究壳聚糖复合保鲜（10 mg/mL壳聚糖＋5 mg/mL茶多酚＋2 000 U/mL溶菌酶）对草鱼冷藏过程中肌肉蛋白质的影响,采用双向电泳-质谱（two-dimensional electrophoresis-mass spectrometry,2DE-MS）技术对新鲜（第0天）和冷藏第9天（保鲜组和未保鲜对照组）草鱼样品进行差异蛋白质组学分析。结果表明,与新鲜样品相比,保鲜组2DE图谱差异不明显,主要变化为快肌钙蛋白T丰度下降,慢肌钙蛋白T和腺苷酸激酶同工酶1丰度上升;而未保鲜对照组样品2DE图谱的蛋白点数量和信号强度均与新鲜组差异较大,进一步分析表明其快肌钙蛋白T、原肌球调节蛋白4、突触足蛋白2、M型丙酮酸激酶丰度下调,肌球蛋白重链、肌球蛋白轻链1、α-肌动蛋白、α-原肌球蛋白、轻酶解肌球蛋白等结构蛋白丰度上升。此外,与保鲜组样品相比,对照组3 种肌钙蛋白T丰度下调,而肌球蛋白重链、轻酶解肌球蛋白、α-肌动蛋白、α-原肌球蛋白、M型肌酸激酶的丰度上升,其中肌酸激酶可作为检测草鱼腐败的指示蛋白。综上,壳聚糖复合保鲜能较好地维持草鱼肌肉蛋白质稳定。     

芬顿反应诱导的肉类蛋白质氧化及干预研究进展

肉及肉制品生产加工过程中伴随着蛋白质氧化的发生,这会导致肉制品营养性和可食用性降低,甚至产生有毒、有害物质,这些有害化合物严重威胁人体健康。蛋白质的氧化修饰改变其结构和理化特性（如构象、结构、组成、溶解性、流变性和反应性）,然而适度的修饰也可起到调控肉制品品质的作用。肉蛋白氧化过程中最主要和最有效的活性氧是羟自由基（·OH）,芬顿反应是产生高活性羟自由基的主要途径之一,因此本文综述芬顿反应中蛋白质氧化的基本原理、反应产物以及蛋白质氧化在肉品加工中对肉类色泽、嫩度、持水性等的作用,以期为肉制品的品质控制提供理论支撑。     

氧化条件下精氨酸对肌原纤维蛋白结构及凝胶性能的调控

以肌原纤维蛋白（myofibrillar protein,MP）为研究对象,以羟自由基作为模拟氧化体系,探究氧化条件下不同浓度L-精氨酸（L-Arg,1、3、5?mmol/L和10?mmol/L）对MP结构及凝胶性能的调控。经不同浓度L-Arg及氧化体系处理后,通过巯基（—SH）及内源色氨酸荧光分析MP构象变化;利用粒径、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE）及溶解度分析MP交联聚集情况;采用流变仪、质构仪及扫描电镜等分析MP凝胶性能;通过拉曼光谱分析MP凝胶二级结构。结果表明：氧化条件下L-Arg对MP结构及凝胶性能的调控具有显著的浓度依赖性：当L-Arg添加量为1～5?mmol/L时,对巯基损失有一定的保护作用,而10?mmol/L时,巯基显著降低。L-Arg的添加使内源色氨酸荧光强度降低、粒度增大,SDS-PAGE分析表明氧化诱导二硫键的形成引起蛋白质交联聚集加剧,进而导致溶解度降低。随添加量增加,MP凝胶微观结构更致密,凝胶强度和白度逐渐降低,但凝胶得率逐渐升高。因此,在氧化条件下,当L-Arg添加量为5?mmol/L时,巯基显著增加,蒸煮损失及凝胶强度显著降低,对肉蛋白的氧化稳定性、肉制品的嫩度和持水性有一定的改善作用。     

发酵和加工对桑椹抗氧化和降血糖作用的影响

目的：研究发酵和加工处理对桑椹抗氧化和降血糖作用的影响。方法：测定桑椹原粉（mulberry powder,MP）、桑椹发酵液（fermented mulberry,FM）、桑椹发酵液冻干粉（freeze-dried fermented mulberry,D（FM））、桑椹发酵液添加黄豆辅料后的冻干粉（freeze-dried fermented mulberry mixed with soybean,D（FM＋S））的单糖含量、菌落总数以及2,2’-联氨-双-3-乙基苯并噻唑啉-6-磺酸（2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate),ABTS）阳离子自由基、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基、羟自由基清除率和亚铁离子螯合能力、α-葡萄糖苷酶抑制率的变化。结果：桑椹经发酵、冷冻干燥处理后,基本检测不到果糖和葡萄糖;D（FM）中肠膜明串珠菌和酵母菌的存活率分别为54.77%、53.40%,而D（FM＋S）中两种菌的存活率都达到90%以上,说明添加黄豆辅料减少了冷冻干燥过程中益生菌的损失。ABTS阳离子自由基清除能力由强到弱依次为D（FM）＞D（FM＋S）＞MP＞FM;DPPH、羟自由基清除率以及亚铁离子螯合能力呈现出一致的趋势：D（FM）和D（FM＋S）清除能力最强,其次为MP,FM最弱。MP、FM、D（FM）、D（FM＋S）对α-葡萄糖苷酶的半抑制浓度分别为0.58、0.53、0.19、0.31 mg /mL,D（FM）和D（FM＋S）对α-葡萄糖苷酶的抑制效果优于MP和FM。结论：桑椹发酵和加工处理得到的冻干粉抗氧化能力和降血糖作用都比FM和未发酵的MP强。     

蛋白质的组成对盐溶蛋白热诱导凝胶特性的影响研究

肌肉盐溶蛋白质形成凝胶的特性是肉制品加工中最重要的功能特性之一,以对虾肉为试材,分别以对虾肉盐溶蛋白热诱导凝胶的硬度、弹性以及黏性为依据,通过聚丙烯酰胺凝胶电泳图谱的比较,得出不同蛋白质的组成对热诱导凝胶特性产生较大的影响,主要的蛋白带出现在31～212ku之间;其组成为肌球蛋白、肌动蛋白、肌原纤维蛋白碎片;电泳图谱中肌球蛋白带宽且颜色深的,其热诱导凝胶的加工特性就越好。     

芦丁诱导猪肉肌原纤维蛋白结构变化对蛋白凝胶特性的改善作用

向猪肉肌原纤维蛋白氧化体系（40?mg/mL蛋白、10?μmol/L?FeCl3、100?μmol/L?VC和1?mmol/L?H2O2）中添加不同量的芦丁（0、10、50、100、150?μmol/g,以蛋白计）,测定蛋白质的巯基含量、表面疏水性、电泳、溶解度、凝胶强度和保水性、流变特性,研究芦丁对肌原纤维蛋白结构和凝胶特性的影响。结果表明,添加芦丁使肌原纤维蛋白的巯基含量显著下降（P＜0.05）,表面疏水性先下降后上升;芦丁使肌球蛋白重链（myosin heavy chain,MHC）强度减弱,肌动蛋白条带强度在其较高含量下（100?μmol/g和150?μmol/g）降低,添加β-巯基乙醇后,MHC和肌动蛋白条带大部分被还原;溶解度随着芦丁添加量的增加而降低;芦丁用量增加,蛋白的凝胶强度与保水性显著增强（P＜0.05）,凝胶最终形成阶段的储能模量（G’）提高。因此,芦丁可通过与肌原纤维蛋白的共价交联或适度增加蛋白的表面疏水性而改善蛋白的凝胶特性。     

姜黄素对羟自由基诱导的肌原纤维蛋白氧化及结构的影响

以猪肉肌原纤维蛋白(MP)为主要研究材料，使用芬顿氧化体系来模拟肉制品的氧化环境，并在其中加入不同浓度(0，5，10，15，20 μmol/L)的姜黄素，通过分析蛋白质氨基酸侧链、二级结构、三级结构以及溶解度的变化，探究不同浓度姜黄素对猪肉肌原纤维蛋白氧化的抑制作用，以及对蛋白质结构的影响。研究结果表明，姜黄素在清除自由基方面表现出良好的效能，能抑制羟自由基引发的蛋白质氧化和结构的改变，并在特定的浓度区间(5~20 μmol/L)内，这种抑制作用与姜黄素的浓度成正比。与氧化组(ox组)相比，姜黄素处理组不仅可以显著抑制羰基的生成和表面疏水性的升高【20 μmol/L的姜黄素处理组对两者的抑制率分别为2.62%，51.78%(P＜0.05)】，也可显著缓解巯基、自由氨基以及溶解度的下降【抑制率分别为80.02%，59.34%，70.84%(P＜0.05)】；添加姜黄素可以促进MP三级结构的展开，对羟自由基引起的二级结构的变化也起到缓解作用。结论:姜黄素可与MP相互作用，从而缓解其氧化变质与结构的变化，对肉制品品质与保鲜有良好的作用。     

羟自由基氧化对鲢鱼肌原纤维蛋白凝胶特性的影响

采用含不同浓度H₂O₂(0.0～10.0 mmol/L)的Fenton体系(H₂O₂-VC-FeCl₃)模拟氧化应激环境对鲢鱼肌原纤维蛋白进行氧化,测定蛋白的巯基含量,以及蛋白凝胶的凝胶强度、白度、持水性、水分分布状态及微观结构,探究氧化度对蛋白凝胶特性的影响。结果表明：随着H₂O₂浓度的增加,蛋白的总巯基含量和白度、持水性呈先升高后降低的趋势。当体系中H₂O₂浓度较低(0.1～0.5 mmol/L)时,蛋白凝胶强度增大,白度得到改善;凝胶的三维结构孔洞的面积较小,且分布较为均匀,凝胶的持水力有所增强。但是当体系中的H₂O₂浓度较高(5.0～10.0 mmol/L)时,蛋白凝胶的白度下降,凝胶结构的有序性降低,孔洞面积变大,分布变得不均匀,凝胶的持水力下降,凝胶品质发生劣变。因此,适度氧化(0.1～0.5 mmol/L H₂O...     

Proteomic analysis of water soluble and myofibrillar protein changes occurring in dry-cured hams

The myofibrillar fraction of raw ham muscles and dry-cured hams with different ripening times was extracted in denaturing and reducing conditions and subjected to two-dimensional gel electrophoresis. The two-dimensional maps gave overall pictures of the already noted progressive disappearance of actin, tropomyosin and myosin light chains during ripening. In addition, two fragments from Myosin Heavy Chain proteolysis, marked as myosin chain fragments MCF1 and MCF2, were identified by immunodetection and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). Furthermore, a new form of actin on two-dimensional gel was identified by MALDI-TOF peptide mapping. In 12-month-old dry-cured ham, most myofibrillar proteins were completely hydrolyzed. At this stage of ripening, in fact, in some Parma and S. Daniele dry-cured ham samples, myosin heavy chain fragments and other unidentified neo-formed spots were found. Some of the sarcoplasmic proteins in water extracts from pork meat markedly decreased in amount or disappeared totally, during ripening. Surprisingly, two-dimensional gel electrophoresis maps of the water soluble protein fraction from dry-cured ham showed the presence of two spots identified as tropomyosin α- and β-chain. This result suggests that some of the saline soluble myofibrillar proteins can disappear from this fraction because of salt solubilization and not due to complete enzyme action. Two-dimensional gel electrophoresis (2-DGE) has proved a powerful tool to evaluate the enzymatic susceptibility of meat proteins and the evolution of protein map fragmentation throughout ripening process as well as a means of obtaining a standard fingerprinting map characterizing the final product.     

过氧自由基对兔肉肌原纤维蛋白理化性质及结构的影响

为探讨脂质氧化对蛋白质氧化的影响,用不同浓度2,2-偶氮二(2-甲基丙基咪)二盐酸盐[2,2′-azobis(2-amidinopropane)dihydrochloride,AAPH]热解产生的脂质初级代谢产物过氧自由基处理兔肉肌原纤维蛋白(myofibril protein,MP)。结果表明,随着AAPH浓度的增加,兔肉MP羰基含量、表面疏水性、亮度值和黄度值显著增加(P<0.05),而MP总巯基、游离氨、溶解度、内源性荧光强度和Zeta电位绝对值都呈显著降低趋势(P<0.05)。拉曼光谱结果显示,MP的α-螺旋含量下降,无规则卷曲含量上升,进一步证实蛋白质二级结构发生改变。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gelelectrophoresis,SDS-PAGE)结果表明,AAPH会导致MP发生交联和聚集,形成分子质量较大的聚集物。因此,过氧自由基会通过促进兔肉肌原纤维蛋白氧化,影响MP的理化性质,研究可为通过调控脂质氧化来控制兔肉加工过程中的蛋白氧化提供一定的理论依据。     

Hydroxyl Radical and Ferryl-Generating Systems Promote Gel Network Formation of Myofibrillar Protein

ABSTRACT: The objective of the study was to examine how oxidatively induced protein cross-linking would influence the gelation properties of myofibrillar protein (MP) under meat processing conditions. MP suspensions in 0.6 M NaCl at pH 6 were treated with an iron-catalyzed oxidizing system (IOS: 10 μM FeCl₃, 0.1 mM ascorbic acid, 0.05 to 5 mM H₂O₂) or a H₂O₂-activated metmyoglobin oxidizing system (MOS: 0.01 to 0.1 mM metmyoglobin/H₂O₂) that produced hydroxyl radical and ferryl species, respectively. Both oxidizing systems promoted MP thermal gelation, which was evidenced by rapid protein–protein interaction and the enhancement in storage modulus (elasticity) of the gel network as revealed by dynamic rheological testing in the 20 to 74 °C temperature range. This gelation-enhancing effect was attributed to the shift of myosin aggregation in the early stage of heating from predominantly head–head association (nonoxidized control samples) to prevalently tail–tail cross-linking through disulfide bonds. However, both hardness and water-holding capacity of chilled gels tended to decline when MP was exposed to ≥1 mM H₂O₂ in IOS and to all concentrations of metmyoglobin in MOS. Microscopic examination confirmed a more porous structure in oxidized gels when compared with nonoxidized protein gels. The results demonstrated that mild oxidation altered the mode of myosin aggregation in favor of an elastic gel network formation, but it did not improve or had a negative effect on water-binding properties of MP gels. Practical Application: Mild oxidation promotes protein network formation and enhances gelation of myofibrillar protein under normal salt and pH conditions used in meat processing. This oxidative effect, which involves disulfide linkages, is somewhat similar to that in bakery product processing where oxidants are used to improve dough performance through gluten protein interaction.     

Evaluation of antioxidant level and protein oxidation of rainbow trout (Oncorhynchus mykiss) fillets during rigor and post-rigor

In this study, the relationship between stunning techniques and protein oxidation which are accepted as the main cause of food spoilage was investigated. For this purpose, the antioxidant status, in vivo myofibrillary protein (MP) oxidation and sensitivity, and postmortem oxidation (inducted with hydroxyl radical system) of rainbow trout (Oncorhynchus mykiss) fillets killed by hitting to head (T1), neck crushing (T2), and convulsion (T3) methods, were investigated. Statistically significant differences (p < .05) were found among all parameters examined and it was observed that the most stressing technique was the convulsion method. It was determined that in protein profiles myosin were influenced too much from stunning technique and in actin observed oxidation-induced reductions. Reductions in S-S and S-H were also found to be increased in carbonyl concentrations, but the most effective values in both processes were determined by the convulsive technique. Our results show that short-term stunning techniques (hitting to head, neck crushing) give better meat quality results in terms of O. mykiss welfare and low MP oxidation rates. In general, we can say that T3 group fillets are more sensitive to oxidative damage, while T1 and T2 groups give better results in maintaining meat quality with low MP oxidation rates.     

Protein Oxidation at Different Salt Concentrations Affects the Cross‐Linking and Gelation of Pork Myofibrillar Protein Catalyzed by Microbial Transglutaminase

In a fabricated then restructured meat product, protein gelation plays an essential role in producing desirable binding and fat‐immobilization properties. In the present study, myofibrillar protein (MFP) suspended in 0.15, 0.45, and 0.6 M NaCl was subjected to hydroxyl radical stress for 2 or 24 h and then treated with microbial transglutaminase (MTGase) in 0.6 M NaCl (E : S = 1 : 20) at 4 and 15 °C for 2 h. Protein cross‐linking and dynamic rheological tests were performed to assess the efficacy of MTGase for mediating the gelation of oxidized MFP. MTGase treatments affected more remarkable polymerization of myosin in oxidized MFP than in nonoxidized, especially for samples oxidized at 0.6 M NaCl. Notably, the extent of MTGase‐induced myosin cross‐linking at 15 °C in oxidized MFP improved up to 46.8%, compared to 31.6% in nonoxidized MFP. MTGase treatment at 4 °C for MFP oxidized in 0.6 M NaCl, but not MFP oxidized in 0.15 M NaCl, produced stronger gels than nonoxidized MFP (P < 0.05). The final (75 °C) storage modulus (G′) of oxidized MFP gels was significantly greater than that of nonoxidized, although the G′ of the transient peak (～44.5 °C) showed the opposite trend. Overall, oxidation at high salt concentrations significantly improved MTGase‐mediated myosin cross‐linking and MFP gelation. This might be because under this condition, MTGase had an increased accessibility to glutamine and lysine residues to effectively initiate protein–protein interactions and gel network formation.