腌制温度对羊肉蛋白质磷酸化水平的影响

研究了不同腌制温度对羊肉肌原纤维蛋白和肌浆蛋白磷酸化水平的影响。取宰后排酸24 h的胴体米龙,设定-1℃、4℃和25℃三个腌制处理组,取腌制0 h、8 h、16 h和24 h的样品,采用SDS-PAGE电泳与荧光染色相结合的方法分析磷酸化水平的变化。结果表明,腌制温度越低,肌原纤维蛋白整体磷酸化水平(P/T值)越高,腌制8 h时,冰温组整体磷酸化水平显著高于室温组(p0.05),腌制16 h和24 h时无显著差异。而肌浆蛋白磷酸化水平结果与之相反,腌制温度越低,肌浆蛋白质整体磷酸化水平越低。腌制8 h时,冰温腌制组整体磷酸化水平显著低于室温腌制组(p0.05),腌制16 h和24 h时也无显著差异。单个蛋白条带呈现差异,随着腌制温度的升高,肌动蛋白和醛缩酶磷酸化水平升高,肌球蛋白和烯醇酶磷酸化水平降低。因此,腌制温度可能通过影响蛋白质磷酸化,进而调控肉的品质。     

宰后成熟过程中羊肉食用品质及蛋白稳定性的变化研究

研究陕北横山羊肉宰后48 h内,不同部位肉块嫩度、持水性等相关指标的变化规律。分别在宰后4、8、12、24、48 h取背最长肌(HQ)和腹部肌肉(LQ),测定了剪切力、汁液流失率和蒸煮损失率、肌浆蛋白和肌原纤维蛋白的溶解度、肌原纤维蛋白降解聚合情况。随着宰后时间延长,两类肉样剪切力在4~8 h显著增大(p0.05),12 h之后显著减小(p0.05);肉样汁液流失率、蒸煮损失率和肌浆蛋白溶解度均显著增加(p0.05);肉样肌原纤维蛋白溶解度4~12 h显著减小(p0.05)而12 h之后逐渐增加(p0.05)。在宰后过程中,HQ肉样剪切力、汁液流失率和蒸煮损失率显著小于LQ肉样(p0.05),肌浆蛋白溶解度显著高于LQ组(p0.05);在宰后12 h内,HQ肉样肌原纤维蛋白溶解度显著低于LQ组(p0.05),但20 h之后反超且显著高于LQ组(p0.05)。两类肉样肌原纤维蛋白的SDS-PAGE电泳结果与溶解度变化趋势一致。宰后2 d内,横山羊肉的嫩度逐渐趋于嫩化,持水性减弱,稳定性减弱;背最长肌羊肉较低品质肉的嫩度和持水性较优、蛋白稳定性较差。     

新疆山羊肉的食用品质及差异蛋白质分析

宰后成熟的过程中,肌肉中蛋白质的表达水平和参与的代谢调节不同,均对肉品质的调节起关键作用。为了深入探索宰后成熟过程中的差异蛋白对新疆山羊肉品质的影响,明确肉质较好的最适成熟时间,该研究测定了宰后不同时间下的新疆山羊背部最长肌的pH值,并采用串联质量标签技术比较死后不同成熟时期的蛋白质水平变化。共得到级质谱谱图总数518 230个,匹配到的谱图总数量为47 666张,唯一肽段总数14 466个,蛋白质总数2 526个。大多数差异表达在山羊宰后成熟中下调表达,其中只有2个糖酵解相关差异蛋白(PGM1,LDHB)上调表达;且与能量代谢相关的线粒体核糖体蛋白L46(MRPL46)、线粒体核糖体蛋白S15(MRPS15)、39S核糖体蛋白L41(MRPL41)、线粒体核糖体蛋白L22(MRPL22)等差异蛋白显著上调表达。结果表明,山羊肌肉的糖代谢过程能维持到肉质成熟的48 h以后,促进肉色的稳定和肉质的嫩化,而线粒体作为能量代谢的关键部位调节肉质嫩度的变化。在成熟期间的肉质逐渐嫩化,且24 h时嫩化程度最高,进一步表明宰后成熟起到嫩度优化的作用。     

AMPK活性对宰后牛肉糖酵解、肌肉内环境及品质的影响

为探究牛肉宰后成熟过程中单磷酸腺苷活化蛋白激酶（adenosine monophosphate-activated protein kinase,AMPK）对糖酵解、肌肉内环境及品质的影响,以0.50 mol/L 5-氨基咪唑-4-甲酰胺核苷（5-amino-4-imidazolecarboxamide,AICAR）处理的西杂牛背最长肌为对象,于4 ℃进行成熟,测定宰后成熟期间肌肉AMPKα基因（PRKAA1、PRKAA2）转录量、P-AMPK表达量、AMPK活性、糖酵解水平及品质指标的变化情况。结果表明：宰后24～120?h,处理组AMPKα基因转录量、P-AMPK表达量及AMPK活力均显著高于对照组（P＜0.05）;72～168?h,处理组pH值和肌糖原含量显著低于对照组（P＜0.05）,乳酸含量显著高于对照组（P＜0.05）;12～168?h,处理组L*、b*值及ATP、ADP、AMP和IMP含量均显著高于对照组（P＜0.05）,a*值显著低于对照组（P＜0.05）;24～120?h,处理组蒸煮损失率和肌原纤维小片化指数显著高于对照组（P＜0.05）,剪切力显著低于对照组（P＜0.05）。AICAR通过激活AMPK并加快宰后糖酵解影响肌肉内环境、肉色、剪切力及肌纤维微观结构变化,加快宰后肌肉成熟进程,说明AMPK活性对宰后肌肉糖酵解及品质变化具有重要影响,且可通过调控宰后肌肉AMPK活性来调节肌肉品质。     

宰后成熟对山羊肉理化性质及蛋白质表达的影响

为探讨山羊宰后48 h内成熟过程中的理化性质规律,分别对宰后0、12、24和48 h山羊背部最长肌的质构特性(TPA)、pH、保水性、肌浆（原）蛋白溶解度以及差异蛋白动态变化等指标进行测定,探讨不同成熟过程中的肉品质变化。结果表明:随着宰后时间点的延长,山羊肉的pH在48 h内显著下降（P<0.05）,且在6.5~5.5之间;滴水流失率在宰后0~24 h显著增加（P<0.05）,但蒸煮损失率却在12 h后下调（P<0.05）;山羊肉成熟中前24 h红度（a*值）与亮度（L*值）显著升高（P<0.05）,而黄度（b*值）却无显著差异（P>0.05）;在宰后12 h内,肌原纤维蛋白溶解度显著下降（P<0.05),随后逐渐增加（P<0.05),而肌浆蛋白溶解度随着宰后时间延长逐渐增大(P<0.05);山羊肉宰后48 h内的糖原含量显著降低(P<0.05)。其中与0 h对比,12 h组的差异表达蛋白总共有13个,24 h共有26个差异表达蛋白,48 h有69个差异表达蛋白,且在0~48 h间能量代谢相关酶上调表达,24~48 h内影响羊肉气味的相关蛋白上调表达。山羊肉在宰后24～48 h的成熟期间,肉质的嫩度、颜色和口感较为优越,此时对山羊肉进行加工和储存,可以为山羊肉的品质提供基础保障。     

禁食时间对宰后早期鸡肉持水力和嫩度的影响

三黄鸡分别禁食0、8、16h和24h,三管齐断法宰杀,于宰后5h内分别测定滴水损失、蒸煮损失、加压损失、蛋白溶解性、剪切力值、糖原、乳酸和pH值等指标,观察禁食对宰后早期鸡肉持水力和嫩度的影响。结果表明：随着禁食时间延长,宰后糖原含量(P＜0.05)和pH值均降低;随着宰后时间延长,糖原含量和pH值变化趋势与宰后时间正相关,而乳酸含量变化趋势与宰后时间负相关。宰后早期,pH值对肌肉持水力有显著影响(P＜0.05),而蛋白溶解度对持水力变化贡献不大。禁食使得宰后早期肌肉剪切力值有增大趋势,但处理组之间差异不显著。与未禁食鸡相比,禁食8h和禁食16h能够提高宰后肌肉持水力。     

蛋白质磷酸化对宰后肉品质影响研究进展

畜禽肌肉在宰后变为可食肉的过程中,需要经过一系列的生化变化直到成熟,其中蛋白质磷酸化反应作为一种蛋白质的动态调控机制,几乎调节每一个主要的生物过程,是生物界常见的翻译后修饰形式之一。因此,研究蛋白质磷酸化与宰后肌肉的关系有助于肉类食品行业的发展。本文从介绍蛋白质磷酸化概念出发,讨论磷酸化蛋白质检测技术的发展,总结宰后肌肉中会发生磷酸化的蛋白质及影响因素,从蛋白质磷酸化反应与宰后肉的嫩度、肉的色泽和保水性3 个方面的研究入手综述蛋白质磷酸化反应对肉品质的影响,以及食盐与氨基酸在该过程中的应用,并对蛋白质磷酸化反应在肉品品质中的研究做出展望。     

氯化钙处理对牛肉嫩度影响的研究

对宰后8h的牛肉分别注射200、250、300mmol／LCaCl2溶液(注射量为肉重的3％),然后将处理样品在4℃下分别腌制12h、24h、48h,通过对其剪切力值的测定,研究注射氯化钙及腌制时间对牛肉嫩度的影响。结果表明,与未注CaCl2溶液组相比,注射CaCl2溶液组牛肉嫩度显著提高(P<0.05),但不同水平CaCl2溶液处理之间牛肉嫩度差异不显著(P>005);不同腌制时间对牛肉嫩度的影响差异不显著(P>0.05)。综合分析,200mmol/L的CaCl2溶液处理浓度、48h的腌制时间改善牛肉的嫩度是可行的。     

宰后僵直温度对肉嫩度的影响

肉的嫩度差和不一致性是肉类屠宰加工工业面临的两大难题。宰后24 h内的胴体/肌肉的温度决定肉嫩度的水平。研究论述了宰后僵直温度对肌肉收缩与成熟过程内源蛋白酶的影响,探讨了宰后温度对肉嫩度影响的机制,进而讨论了p H-温度窗口,以期为屠宰加工企业优化胴体冷却温度提供参考。     

宰后早期猪肉、牛肉和鸡肉中能量代谢及蛋白质磷酸化

目的:探讨畜禽肉宰后能量代谢的差异及其可能的原因。方法:于宰后45 min测定鸡肉、灰白(pale soft exudative,PSE)猪肉、正常猪肉和牛肉中糖原、ATP/ADP/AMP含量及乳酸脱氢酶(lactate dehydrogenase,LDH)活性,并提取肌浆和肌原纤维蛋白进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfatepolyacrylamide gel electrophoresis,SDS-PAGE)和液相色谱-串联质谱鉴定,分析各组肉的磷酸化水平差异。结果:牛肉中糖原含量显著高于其他组(P0.05),正常猪肉组糖原和ATP含量显著高于PSE猪肉组和鸡肉组(P0.05),而PSE猪肉组的LDH活性显著高于其他组(P0.05);肌浆蛋白SDS-PAGE ProQ染色表明,宰后45 min PSE猪肉组中6个条带的磷酸化水平显著低于正常猪肉组(P0.05),而另有6个条带的磷酸化水平显著高于正常猪肉组(P0.05)。牛肉组和鸡肉组中分别有3个和1个条带的磷酸化水平偏低,1个条带和8个条带的磷酸化水平明显偏高。而肌原纤维蛋白电泳表明,牛肉中大部分蛋白质的磷酸化水平处于最高值,而鸡肉组处于最低值,猪肉介于两者之间;PSE猪肉组中7个条带的磷酸化水平显著低于正常猪肉(P0.05);6个条带的磷酸化水平显著高于正常猪肉(P0.05)。结论:宰后异质肉的形成受宰后糖原和ATP消耗速率、LDH活性的影响,同时与肌肉中糖酵解酶的磷酸化水平有关。     

Alterations in the sarcoplasmic protein fraction of beef muscle with postmortem aging and hydrodynamic pressure processing

Abstract: Capillary electrophoresis (CE) and reversed‐phase high performance liquid chromatography (RP‐HPLC) analyses were utilized to detect differences in the sarcoplasmic protein fractions of beef strip loins subjected to aging and hydrodynamic pressure processing (HDP) treatments. At 48 h postmortem, strip loins (n= 12) were halved and subjected to control or HDP treatments. Following treatment, each half was divided into 3 portions which were aged for 0, 5, and 8 d. After each aging period, steaks were removed for Warner–Bratzler shear force (WBSF) analysis and for the extraction of sarcoplasmic proteins which were analyzed by CE and RP‐HPLC. Aging by HDP interactions were not detected using either separation technique. With CE analysis, no HDP effects were observed; however, the relative peak area of 8 protein peaks ranging in size from 17 to >200 kDa were influenced by postmortem aging. Separation of proteins by RP‐HPLC demonstrated that HDP influenced the relative size of 2 protein peaks while postmortem aging effects were observed in 6 peaks. Alterations in the sarcoplasmic protein fractions detected by both CE and RP‐HPLC were correlated to WBSF measurements. Overall, data demonstrate that HDP has minimal effects on sarcoplasmic proteins and that aging related changes in the water soluble protein fractions of muscle may be useful as indirect indicators of beef tenderness. Practical Application: Using 2 different postmortem tenderization techniques, aging and hydrodynamic pressure processing, this study demonstrates that postmortem changes in the soluble protein fraction of beef may be useful as potential indicators of meat tenderness.     

活性氧激活缺氧诱导因子-1α加速宰后成熟初期牛肉能量代谢

为探究活性氧（reactive oxygen species,ROS）在宰后成熟初期调控牛肉缺氧诱导因子-1α(hypoxiainduciblefactor-1α,HIF-1α)对能量代谢的影响。牛肉样品分别用H2O2、N-乙酰半胱氨酸（N-acetyl-L-cysteine,NAC）和生理盐水（对照）处理。在低温成熟0.5、6、12、24、48 h分别测定ROS含量、脯氨酰羟化酶（proline hydroxylase,PHD）活力、磷酸肌醇-3激酶（phosphatidylinositol3-kinase,PI3K）/蛋白激酶B(proteinkinase B,AKT)和HIF-1α蛋白质表达水平、能量代谢水平以及pH值的变化。结果发现：H2O2处理组初始（0.5 h）ROS含量显著高于其他2组（P<0.05）,且在48 h内始终处于显著较高水平（P<0.05）,NAC处理能够在短期内（0.5～12 h）抑制ROS积累;H2O2处理组PHD活力在0.5～48 h内显著低于其他2组（P<0.05）,同时,在0.5～12 h和0.5～6 h内PI3K和磷酸化AKT表达...     

宰后僵直及成熟过程中羊背最长肌理化性质的变化

动物宰后从肌肉到肉品经过僵直、成熟等一系列复杂的生理生化反应。僵直及成熟过程的研究可为肉品质改善及肉制品加工提供理论依据。选取杂交公羊(小尾寒羊×北京本地羊)双侧背最长肌在4℃成熟0.5、2.0、6.0、12.0、24.0、48.0、72.0、120.0、168.0 h,测定不同时间的pH值、剪切力、肌节长度、ATP含量、肌原纤维小片化指数(myofibril fragmentation index,MFI)、钙蛋白酶活力。结果表明:宰后成熟过程中,pH值先下降后逐渐趋于稳定,从第24小时后变化不显著(P0.05);剪切力先上升后下降,在第24小时达到最大值;肌节长度在宰后先缩短后逐渐变长,且在第48小时缩至最短;ATP含量先上升后下降,48 h后趋于稳定;μ-钙蛋白酶80 kD大亚基在宰后24 h基本降解完全,其降解的78 kD大亚基在第48小时降解完全;肌间线蛋白和肌钙蛋白T作为μ-钙蛋白酶的降解底物,在成熟过程中发生降解,第168小时几乎观察不到完整的蛋白条带;随着宰后时间的延长,羊背最长肌从第2小时开始僵直,到第24小时程度达到最大,从第48小时开始解僵,解僵后肉的嫩度逐渐改善。     

Difference in tenderness and pH decline between water buffalo meat and beef during postmortem aging

The objective of this research was to determine the difference in tenderness and some characteristics of water buffalo meat and beef during postmortem aging. Five female crossbred water-buffalo (Philippine Carabao×Bulgarian Murrah) and five female crossbred cattle (Brahman×Philippine Native), were finished on the same diet for 6 months and slaughtered at 30 months of age. The muscle pH was measured at 40min, 3h, 7h, 24h, and 48h postmortem. Longissimus thoracis (LT) and semimembranosus (SM) muscles were excised at 2d postmortem, and shear force was measured at 2, 4, 7, and 14d postmortem. Glycogen and lactate concentrations were determined from 0, 2, and 4d LT samples, and myosin heavy chain type of buffalo and cattle LT was determined by ELISA methods. Myofibrillar protein degradation was also observed by SDS-PAGE and Western blotting of fast-type troponin T. Results showed that the buffalo meat had significantly lower shear force values compared to beef for LT and SM muscles, which was supported by a difference in troponin T degradation. Postmortem pH decline of buffalo meat was significantly slower than that of beef, which was confirmed by lactic acid concentrations, but was not explained by glycogen content. In addition, there was no significant difference in the ratio of slow to fast type muscle fibers in buffalo and cattle, indicating that myosin heavy chain type was not responsible for the difference in pH decline and tenderness between the buffalo meat and beef. This study demonstrated that the tenderness of water buffalo meat was superior to that of Brahman beef, which may have been due to the difference in pH decline and the subsequent effect on muscle protease activity.     

包装方式和宰后不同时间包装对羊肉品质的影响

为探究包装方式和宰后不同时间包装对羊肉贮藏品质的影响,本研究对宰后1 h热分割羊肉和冷却成熟24 h后的分割羊肉分别进行热收缩包装、真空包装、贴体包装、50%（体积分数,下同）O2＋50% CO2气调包装,测定其在（2±2）℃贮藏过程中pH值、色泽、贮藏损失率、蒸煮损失率、离心损失率、剪切力、挥发性气味的变化。结果表明：与宰后24 h包装相比,宰后1 h包装羊肉pH值在贮藏14 d时更高,但其贮藏期间色泽较好,色泽稳定性更高,贮藏损失率更低。从整体来看,3 种真空处理（热收缩包装、真空包装、贴体包装）羊肉嫩度均优于气调包装羊肉,且宰后1 h包装时真空条件处理对羊肉嫩化效果最好,宰后24 h羊肉在贮藏前期气调包装对羊肉嫩化效果较好;而无论何时包装,热收缩包装和气调包装羊肉贮藏损失率均显著小于其他处理组（P＜0.05）,在贮藏前期色泽及稳定性较好,但贮藏后期色泽稳定性下降明显,气调包装羊肉在14 d时已部分发绿,贮藏21 d时挥发性气味变化显著。综上,宰后1 h包装有利于提高羊肉贮藏期间色泽及持水力,改善真空处理羊肉嫩度,宰后24 h包装可提高50% O2＋50% CO2气调包装羊肉贮藏前期的嫩度;其中热收缩包装和气调包装羊肉持水力高,贮藏前期色泽及稳定性好,真空包装、贴体包装羊肉品质变化相对稳定,适宜长期贮藏。     

Effects of Prerigor Pressurization on the Emulsifying Capacity of Muscle Protein

The emulsifying capacities of pressure treated and control muscle homogenates, sarcoplasmic protein and myofibrillar proteins of ovine and bovine longissimus muscles were determined at 2, 6, 24 and 168 hr postmortem. The pH of the intact muscle, muscle homogenate and myofibrillar protein extract were taken at these times. Before onset of rigor mortis, the emulsifying capacity of muscle homogenate from the control samplcs was higher than the pressure treated samples. At 24 and 168 hr postmortem, the pressure treated and control samples were not significantly different (P>0.05) for emulsifying capacity. At 2 hr postmortem, the emulsifying capacity of myofibrillar protein extract from control samples was higher (P<0.05) than that from pressure treated samples; there- after, the emulsification curve for the pressure treated samples was higher than that of the control. The emulsification capacity of sarcoplasmic proteins from control muscles was slightly, but consistently, higher than that from pressure treated muscles throughout the test period. Overall, the emulsification capacity of muscle proteins was not detrimentally affected by pressure treatment.     

宰后成熟对牛胴体不同部位显微结构影响

将不同部位肉样真空包装后于4℃条件下成熟72h ,采用石蜡切片苏木精-伊红(HE)染色法、原子力显微镜扫描肌原纤维、环境扫描电镜扫描肌纤维膜、免疫组织化学法及测定剪切力,研究牛胴体不同部位剪切力、宰后成熟过程中肌纤维和肌原纤维裂解程度、肌纤维膜变化及μ-calpain肌纤维定位。结果表明：剪切力：半腱肌＞腹直肌＞背最长肌;牛肉后熟过程中,背最长肌肌纤维、肌原纤维裂解程度最大,腹直肌次之,半腱肌变化较小;肌纤维膜变化不大;μ-calpain主要存在于肌纤维膜和肌浆中,红肌纤维(慢肌)μ-calpain含量高,白肌纤维(快肌)μ-calpain含量低。μ-calpain在牛肉后熟过程中对牛肉嫩化具有重要作用,红肌纤维μ-calpain含量高,白肌纤维μ-calpain含量低,因此,胴体不同部位红肌纤维和白肌纤维的比例是导致其后熟嫩度和显微结构差异较大的一个重要因素。     

Impact of carcass scalding and chilling on muscle proteins and meat quality of broiler breast fillets

The objective of this study was to determine the effects of broiler carcass scalding and chilling methods on meat quality and muscle proteins. During processing, carcasses were hard (60 °C, 1.5 min) or soft (52.8 °C, 3 min) scalded, and either immersion chilled (IC: 0.5 °C, 40 min) or air chilled (AC: 0.5 °C, 120 min). Breast fillets were deboned at 4 h postmortem and used for measuring meat quality and muscle protein characteristics. Scalding by chilling treatment interaction effects on meat quality were not observed. Air chilled carcasses had greater pH₂₄, and reduced drip loss and shear force compared to IC carcasses. Cook yield, color (L*a*b*), and moisture content were not different between chilling treatments. Scalding treatments did not influence quality traits. Sarcoplasmic protein solubility was not influenced by chilling treatment, but was greater in hard versus soft scalded carcasses. Myofibrillar protein solubility was greater in fillets from soft scalded IC carcasses. Alterations in the electrophoretic profiles of the myofibrillar and sarcoplasmic proteins due to treatments indicated minor changes in protein degradation and solubility. Data suggest that while only chilling method influenced meat quality, both scalding and chilling methods influenced protein solubility and degradation in breast fillets deboned 4 h postmortem.     

高强度超声对鹅胸肉嫩度及品质的影响

为了解超声技术对鹅胸肉品质的影响,采用高强度超声（超声功率800 W,超声总时间42 min,工作时间 2 s、停歇时间3 s）对鹅胸肉进行处理。测定未超声和超声处理的鹅胸肉于4 ℃下放置不同时间（0、6、12、24、 36、48 h）的pH值、肌原纤维小片化指数、蒸煮损失率、剪切力、表面微观结构、肌原纤维蛋白降解情况、肉品 的热力学性质等指标。结果显示：与未经超声处理的鹅胸肉相比,超声处理后随着放置时间的延长,肉品pH值升 高,肌原纤维小片化指数显著增大,蒸煮损失率和剪切力显著减小（P＜0.05）;表面微观结构发生明显变化,肌 动蛋白和肌球蛋白热敏性增大,导致更多肌原纤维蛋白降解,游离肌动蛋白含量显著增加（P＜0.05）,从而显著 改善鹅胸肉的嫩度。因此,超声处理可破坏鹅胸肉肌原纤维的完整结构,缩短宰后肉品成熟所需时间,进一步提高 鹅胸肉的品质。