Monitoring Proteolysis During Cheddar Cheese Ripening Using Two-Dimensional Gel Electrophoresis

A 2-D gel electrophoretic method, consisting of isoelectric focusing and alkaline urea-PAGE was used to monitor proteolysis during ripening (180d, 5°C and 8°C) of full- and reduced-fat Cheddar cheese. The method enabled quantifying changes in levels of peptides in cheese with good spot-resolution. Results can complement those from other analyses, especially those for determining low MW peptides. Notable effects were found for cheese composition and ripening temperature on gel pattern and on relative levels of selected proteolysis products. In both cheese varieties, most peptides reached a maximum during the first 3 ripening months and gradually disappeared as ripening advanced.     

Metformin Attenuates Slow-to-Fast Fiber Shift and Proteolysis Markers Increase in Rat Soleus after 7 Days of Rat Hindlimb Unloading

Muscle unloading leads to signaling alterations that cause muscle atrophy and weakness. The cellular energy sensor AMPK can regulate myofiber-type shift, calcium-dependent signaling and ubiquitin-proteasome system markers. We hypothesized that the prevention of p-AMPK downregulation during the first week of muscle unloading would impede atrophy development and the slow-to-fast shift of soleus muscle fibers, and the aim of the study was to test this hypothesis. Thirty-two male Wistar rats were randomly assigned to four groups: placebo control (C), control rats treated with metformin (C + M), 7 days of hindlimb suspension (HS) + placebo (7HS), and 7 days of HS + metformin administration (7HS + M). In the soleus of the 7HS rats, we detected a slow-to-fast fiber-type shift as well as a significant downregulation of MEF-2D and p300 in the nuclei. In the 7HS group, we also found decreases in p-ACC (AMPK target) protein level and in the expression of E3 ubiquitin ligases and p-CaMK II protein level vs. the C group. The 7-day metformin treatment for soleus muscle unloading (1) prevented slow-to-fast fiber-type shift; (2) counteracted changes in the p-ACC protein level; (3) hindered changes in the nuclear protein level of the slow myosin expression activators MEF-2D and p300, but did not affect NFATc1 signaling; and (4) attenuated the unloading-induced upregulation of MuRF-1, atrogin-1, ubiquitin and myostatin mRNA expression, but did not prevent soleus muscle atrophy. Thus, metformin treatment during muscle disuse could be useful to prevent the decrease in the percentage of slow-type fatigue-resistant muscle fibers.     

Alcalase对高沟捆蹄肌肉蛋白质水解变化规律研究

以猪后蹄肉为原料,在腌制过程添加Alcalase作为酶解剂,并通过响应曲面实验法优化酶解工艺;通过SDSPAGE电泳分析及游离氨基酸组分分析Alcalase对捆蹄蛋白质的酶解作用规律。结果表明,Alcalase在腌制温度为4~16℃时,对促进高沟捆蹄蛋白质水解及提高其感官品质有显著的促进的作用(p<0.05),过量的Alcalase(大于1.6 U/g)会导致蛋白质过度水解,不利于产品的感官品质。响应曲面法优化捆蹄加工工艺结果为:Alcalase添加量1.32 U/g、腌制温度14.2℃、腌制时间21.3 d,产品蛋白质水解指数为14.81%,感官评分92.47。另外,添加Alcalase蛋白酶后,捆蹄中高分子蛋白(≥120 ku)有明显的分解作用,并且在30~40 ku及50~70 ku处,蛋白质条带逐渐累积;Ala、Leu、Val、Tyr、Phe、Ile、Gly、His有显著的增加,而Pro、Glu含量显著降低(p<0.05)。因此,Alcalase能显著促进捆蹄中蛋白质的分解,提高产品的感官品质,可以应用于捆蹄产业的生产加工。      

Impact of dietary fibers on the properties and proteolytic digestibility of lactoferrin nano-particles

The application and development of biopolymer-based particulate delivery systems is receiving considerable attention from scientists and manufacturers seeking to engineer healthier and safer foods. Present work focused on the fabrication and proteolysis of bovine lactoferrin nano-particles onto which pectins, alginate and carrageenan were electrostatically deposited. DLS, CD and AFM were used to ascertain the formation of spherical structures with varying zeta potentials and modified protein folding. SDS-PAGE analyses of samples collected during simulated human gastro-duodenal digestion revealed that in the case of high and low methoxy pectin particle zeta potential values (−35.4 mV and −50.3 mV respectively) were found to be inversely linked to lactoferrin digestion rate. Moreover, the electrostatic binding of food grade iota-carrageenan onto lactoferrin nano-particles was found to yield highly charged particles (zeta = −69.2 mV) with low surface roughness and to enable a proportion of the lactoferrin to resist 1 h of simulated gastric digestion. Overall, these findings suggest that electrostatic biopolymer interactions may modulate protein digestion and possibly facilitate controlled protein delivery to the upper gastrointestinal tract or have implications to protein nutritional value.     

甲醇芽孢杆菌凝乳酶对马苏里拉干酪加工特性的影响

为了探究甲醇芽孢杆菌（Bacillus methanolicus）凝乳酶在马苏里拉干酪加工中的应用,分别以使用甲醇芽孢杆菌凝乳酶、混合酶制剂（含质量分数10%甲醇芽孢杆菌凝乳酶和90%商品凝乳酶）制作的马苏里拉干酪作为实验组,以商品凝乳酶干酪作为对照组,测定不同组别干酪成熟期间的蛋白水解特性、质构、风味和微观结构变化,研究甲醇芽孢杆菌凝乳酶对马苏里拉干酪加工特性的影响。结果表明,实验组干酪在成熟过程中pH值（4.6～5.3）、微生物数量（8.80～9.68（lg（CFU/g）））与对照组无显著差异（P＞0.05）;实验组干酪水分质量分数（混合酶干酪为（43.21±1.17）%、甲醇芽孢杆菌凝乳酶干酪为（46.15±0.94）%）均显著高于对照组（（41.08±1.04）%）,得率（混合酶干酪为（9.27±0.17）%、甲醇芽孢杆菌凝乳酶干酪为（9.46±0.16）%）也显著高于对照组（（8.98±0.13）%）（P＜0.05）;且实验组干酪的蛋白水解特性（pH 4.6时可溶性蛋白、酪蛋白水解程度和游离氨基酸质量分数）以及风味物质种类和相对含量等指标也优于对照组干酪。但是实验组中甲醇芽孢杆菌凝乳酶干酪保形性相对欠佳,感官评定得分偏低,混合酶干酪与对照组质构及感官基本得分一致,因此甲醇芽孢杆菌凝乳酶可以作为商品酶的部分代替品应用于干酪的生产中。     

Effect of the fungal protease EPg222 on proteolysis and texture in the dry fermented sausage ‘salchichón’

Proteolytic activity of the fungal protease EPg222 and its effect on the texture of the dry fermented sausage ‘salchichón’, which has a long ripening process, was assayed. Samples with enzyme added showed a significant reduction of myofibrillar protein concentration during the ripening period, compared with a control, and proteolytic activity of the enzyme led to a higher accumulation of non‐protein and ‐amino acid nitrogen. Analysis of volatile compounds in ripened dry fermented sausages revealed that only in EPg222 batches were branched compounds derived from amino acids catabolism detected that are associated with the flavour of dry cured meat products. The texture profile analysis showed reduction in hardness, gumminess and chewiness values in treated compared with control sausages. The effect of this enzyme could be of great interest in stimulating proteolysis, in flavour development and in reducing the hardness of dry fermented sausages produced by a long ripening process. Copyright © 2004 Society of Chemical Industry     

碱性蛋白酶水解对豆粕中大豆抗原蛋白的影响

大豆是八大食物过敏源之一,其中主要蛋白成分7S和11S球蛋白也是致敏性较强的抗原蛋白。本试验研究了豆粕在Alcalase酶水解过程中7S和11S两种大豆抗原蛋白的变化。结果表明,酶解10 min时7S伴球蛋白的α’、α和β亚基、酶解60 min时11S球蛋白的酸性亚基等主要抗原蛋白成分即被完全水解,同时新产生了23 ku和大量14 ku以下组分;酶解豆粕中95.1%的蛋白可溶于水,其中68.1%蛋白质可溶于三氯乙酸。     

我国广式腊肠研究进展

广式腊肠是我国一种传统腌腊肉制品,近年来,其质量安全问题越来越受到人们的关注,关于广式腊肠的相关研究也日益广泛和深入。本文对广式腊肠的质量特性、加工工艺、配方的改进以及加工过程中的生理生化变化规律及作用机理等研究进展进行综述,旨在为其今后的研究提供参考。