Serine Protease Inhibitors as Good Predictors of Meat Tenderness: Which Are They and What Are Their Functions?

Since years, serine proteases and their inhibitors were an enigma to meat scientists. They were indeed considered to be extracellular and to play no role in postmortem muscle proteolysis. In the 1990's, we observed that protease inhibitors levels in muscles are a better predictor of meat tenderness than their target enzymes. From a practical point of view, we therefore choose to look for serine protease inhibitors rather than their target enzymes, i.e. serine proteases and the purpose of this report was to overview the findings obtained. Fractionation of a muscle crude extract by gel filtration revealed three major trypsin inhibitory fractions designed as F1 (Mr:50–70 kDa), F2 (Mr:40–60 kDa) and F3 (Mr:10–15kD) which were analyzed separately. Besides antithrombin III, an heparin dependent thrombin inhibitor, F1 and F2 comprised a large set of closely related trypsin inhibitors encoded by at least 8 genes bovSERPINA3-1 to A3-8 and able to inhibit also strongly initiator and effector caspases. They all belong to the serpin superfamily, known to form covalent complexes with their target enzymes, were located within muscle cells and found in all tissues and fluids examined irrespective of the animal species. Potential biological functions in living and postmortem muscle were proposed for all of them. In contrast to F1 and F2 which have been more extensively investigated only preliminary findings were provided for F3. Taken together, these results tend to ascertain the onset of apoptosis in postmortem muscle. However, the exact mechanisms driving the cell towards apoptosis and how apoptosis, an energy dependent process, can be completed postmortem remain still unclear.     

Comparative studies of quality changes in white and dark muscles from common carp (Cyprinus carpio) during refrigerated (4 °C) storage

Differences between white and dark muscles from common carp (Cyprinus carpio) in sensory assessment, total viable counts (TVC), total volatile basic nitrogen (TVB‐N), pH, electrical conductivity (EC), biogenic amines (BAs), nucleotide degradation products and colours were investigated under air and vacuum conditions during refrigerated (4 °C) storage. Compared with white muscle, dark muscle had a higher lipid content, but lower crude protein and moisture content. TVC, TVB‐N, pH, and L* value were higher in white muscle, while K value and a* value were higher in dark muscle. Under both conditions, white and dark muscles had significant (P < 0.05) differences in tyramine and cadaverine content. Although dark muscle was significantly (P < 0.05) lower in ATP content, it had higher rate of IMP degradation and K value increase. Taking sensory and microbial indexes into consideration, dark muscle appeared to have a longer shelf life than white muscle under both conditions.     

Changes in apoptosis factors and activation of caspase-3 in tilapia muscle during storage

The activation of the apoptosis pathway in tilapia muscle during postmortem storage was studied. Changes in caspase-3 activity, ATP content, cytochrome c levels, and ratio of Bcl-2/Bax levels of tilapia muscle were observed during postmortem storage at 20°C. Caspase-3 activity was found to be significantly increased at first, followed by a decrease (P < 0.05); the highest caspase-3 activity was observed at 1 h. The ATP content decreased significantly (P < 0.05), and almost exhausted after 10 h storage. The cytochrome c level in the cytosol showed a significant increase after 5 h of storage (P < 0.05), while the mitochondrial cytochrome c levels showed a decrease. The Bcl-2/Bax ratio was stable from 0–5 h, followed by a rapid decreased at 10–20 h and a significant increased after 20 h (P < 0.05), suggesting that the apoptosis process occurred until 20 h of postmortem storage. Thus, we concluded that the availability of ATP and the increase in cytosolic cytochrome c levels are essential for the activation of caspase-3, and that the former partly limits caspase-3 activity.     

The assessment of carcass composition of Awassi male lambs by real-time ultrasound at two different live weights

The accuracy of ultrasonography to measure fat thickness (FT) and longissimus dorsi muscle (LM) traits (area, depth and width) in live Awassi male lambs and predict carcass FT and LM traits was studied.Twenty six Awassi male lambs were randomly divided into light (L) (n = 13) and heavy (H) (n = 13) finishing lambs. Slaughter weight of lambs in L and H groups were 40 and 45 kg, respectively. FT and LM traits, cross-sectional area between the 12th and 13th rib were measured using real-time ultrasound in vivo and on the carcass after slaughter. All ultrasound and carcass measurements were the same except live weight (LW) (P < 0.001), cold carcass weight (CCW) (P < 0.001) and carcass LM width (P < 0.05). Overall, correlation coefficients between ultrasound and carcass FT, LM depth, width and area were 0.79, 0.82 (P < 0.001); 0.60, 0.58 (P < 0.05); 0.48, −0.17 (P > 0.05) and 0.89, 0.87 (P < 0.001), respectively, for lambs in L and H groups. The introduction of ultrasound FT and ultrasound LM area as independent variables in addition to LW in the multiple regression equations further improved the variations for carcass FT (84%, 71%), carcass LM area (79%, 79%), CCW (72%, 65%) for lambs in light and heavy groups whereas no improvement was observed for carcass yield.These results indicate that in vivo ultrasound FT and measurement of the LM area in association with LW could be used to estimate carcass FT, carcass LM area and CCW in different LW Awassi lambs.     

SDS-PAGE法检测动物肌肉蛋白质加热终点温度的研究

为了探索检测动物组织蛋白质热变性程度的方法,分别对猪、牛、羊、鸡、鱼五种新鲜动物肌肉组织进行不同温度的热处理（新鲜未加热、50、60、70、80、90、100℃）,对处理后的样品提取蛋白质进行十二烷基磺酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析。结果显示,同种动物组织经不同温度热处理,所得电泳图谱具有较大差异,电泳条带的数量随温度的升高逐渐减少,猪、牛、羊三种动物肌肉组织加热到80℃后,电泳图谱不再变化,达到加热终点温度;鸡和鱼的肌肉组织加热到70℃后,电泳图谱不再变化,达到加热终点温度。不同种动物组织经相同温度热处理所得电泳图谱显示：猪、牛、羊三种动物组织蛋白质的电泳图谱相似,与鸡和鱼组织蛋白质的电泳图谱差异显著。     

野生与池养刀鲚肌肉品质特性及抗氧化性的比较分析

比较研究了野生和池养刀鲚（Coilia nasus）肌肉营养成分组成、质构特性和抗氧化特性。结果显示,野生刀鲚肌肉中水分、羟脯氨酸、胶原蛋白含量以及胶原蛋白/总蛋白比值均显著低于池养型,而脂肪含量则显著提高;两者的蛋白质含量、pH均无显著性差异;池养刀鲚肌肉弹性、硬度、咀嚼度、胶着性相较于野生型呈上升趋势,但无统计学差异。野生刀鲚肌肉中SOD、CAT、T-AOC、MDA、GSH-PX、iNOS、GSH/GSSG均显著高于池养型。表明野生和池养刀鲚营养品质存在差异,建议在人工养殖刀鲚中应该注意饵料的种类和丰度,其养殖模式有待进一步改善。     

合浦珠母贝肉酶解液中抗氧化肽的分离及活性研究

合浦珠母贝肉酶解液经膜分离后,采用葡聚糖凝胶G-25进行分离纯化,对分离组分的总抗氧化性、DPPH自由基清除能力、羟自由基清除能力和在化妆品中的应用效果进行测定,结果表明:经分离得到10个有效组分F1-F10,其中F4组分的总抗氧化性最强,DPPH自由基清除能力44.1±0.3%,羟自由基清除率76.0±1.5%,将合浦珠母贝抗氧化肽(F4组分)添加到营养霜中,当添加量为1.0%时,抗氧化活性A700nm为1.599±0.004,能有效提高营养霜的抗氧化性,且营养霜的性状没有发生改变。该研究为合浦珠母贝肉抗氧化肽的结构和在化妆品中的应用研究奠定基础。     

瓜拉纳抗疲劳功效的研究

雄性昆明小鼠按体重随机分为3组(I-瓜拉纳、II-西洋参液、III-纯水对照)。每组小鼠每日分别一次性灌服剂量20mL/kg,连续灌胃35d后,进行耗竭性游泳实验,测定小鼠游泳时间,运动后全血乳酸水平、血清尿素氮水平及肝糖原含量。结果表明:与纯水相比,瓜拉纳、西洋参可显著降低雄性小鼠体内的血乳酸水平,其中瓜拉纳还能显著提高雄性小鼠体内的肝糖原储备。     

植物乳杆菌6003对肌肉蛋白质的分解能力(Ⅰ)

本文研究了植物乳杆菌6003活细胞(Cell)、细胞裂解液(Cellextracts,CE)以及两者混合液(Cell+CE)对肌浆提取液中的蛋白质的分解作用。通过蛋白质、肽、游离氨基酸分析,表明Cell组以及Cell+CE组对肌浆提取液中蛋白质、肽分解和游离氨基酸的增加均有一定作用,其中Cell+CE组中这一现象更为显著。CE组对肌浆蛋白质的分解不起作用。     

Antiproliferative activity of peptides prepared from enzymatic hydrolysates of tuna dark muscle on human breast cancer cell line MCF-7

To produce and identify antiproliferative peptides, two commercial enzymes, papain (PA) and Protease XXIII (PR) were used to hydrolyse tuna dark muscle byproduct, and the protein hydrolysates were purified, before being evaluated for antiproliferative activities against human breast cancer cell line MCF-7. The results showed that the peptide fractions with the molecular weight ranging from 390 to 1400 Da possessed the greatest antiproliferative activity. The amino acid sequences of the two antiproliferative peptides isolated from PA and PR hydrolysates were Leu-Pro-His-Val-Leu-Thr-Pro-Glu-Ala-Gly-Ala-Thr (1206 Da) and Pro-Thr-Ala-Glu-Gly-Gly-Val-Tyr-Met-Val-Thr (1124 Da), whilst they show the dose-dependent inhibition effect of the MCF-7 cells with IC₅₀ values of 8.1 and 8.8 μM, respectively. We thus conclude that antiproliferative hydrolysates from tuna dark muscle byproduct may be useful ingredients in food and nutraceutical applications.