Autolysis and the endogenous proteinases characterised in beardless barb (Anematichthys apogon) muscle

Beardless barb is a common fish species used in fermentation of fish paste Ka-pi-plaa. Autolytic profile of beardless barb muscle showed the maximum autolysis was at 50 °C, at both acidic and alkaline pH values. With augmentation concentration of NaCl, autolytic activity slightly decreased. Endogenous proteinases isolated from fish muscle in crude extract forms were also characterised. The acidic proteinases had optimum activity at pH 3.0 and 50°C, and they showed higher proteolytic activity than the alkaline proteinases which were optimally active at pH 9.0 and 50 °C. Proteinases in peak at pH 3.0 were inhibited by pepstatin A, but those in peak at pH 9.0 were highly inhibited by PMSF, TLCK and soybean trypsin inhibitor, suggesting that both aspartic and serine proteinases were existed in beardless barb muscle. The proteinases were stable in pH range of 2.0-5.0 but unstable at the temperatures higher than 40 °C. NaCl suppressed the proteolytic activity, ATP activated the proteinase activity, while CaCl₂, MgCl₂ and CoCl₂ exhibited no influence on the activity. The results implied that cathepsin D is the predominant proteinase responsible for autolysis in beardless barb. The findings were useful to improve the processing and qualities of Ka-pi-plaa product using beardless barb as raw material.     

The Potential Role of Kallistatin in the Development of Abdominal Aortic Aneurysm

Abdominal aortic aneurysm (AAA) is a vascular condition that causes permanent dilation of the abdominal aorta, which can lead to death due to aortic rupture. The only treatment for AAA is surgical repair, and there is no current drug treatment for AAA. Aortic inflammation, vascular smooth muscle cell apoptosis, angiogenesis, oxidative stress and vascular remodeling are implicated in AAA pathogenesis. Kallistatin is a serine proteinase inhibitor, which has been shown to have a variety of functions, potentially relevant in AAA pathogenesis. Kallistatin has been reported to have inhibitory effects on tumor necrosis factor alpha (TNF-α) signaling induced oxidative stress and apoptosis. Kallistatin also inhibits vascular endothelial growth factor (VEGF) and Wnt canonical signaling, which promote inflammation, angiogenesis, and vascular remodeling in various pre-clinical experimental models. This review explores the potential protective role of kallistatin in AAA pathogenesis.     

壳聚糖固定瑞士乳杆菌蛋白酶的条件研究

以壳聚糖为载体,戊二醛为交联剂,采用交联-吸附法对瑞士乳杆菌蛋白酶的固定化条件进行研究。在单因素试验基础上,以固定化酶活力为主要指标,研究凝结液、壳聚糖质量浓度、酶用量、交联时间、戊二醛质量浓度对瑞士乳杆菌蛋白酶固定化的影响。运用响应面对固定化条件进行优化,确定瑞士乳杆菌蛋白酶的最优固定条件：凝结液为4g/100mL NaOH-甲醇(体积比3:1)、壳聚糖质量浓度2.89g/100mL、酶用量2.95mg、交联时间1h、戊二醛质量浓度0.40g/100mL,此时固定化酶活力为28.67U。     

High γ-aminobutyric acid production from lactic acid bacteria: Emphasis on Lactobacillus brevis as a functional dairy starter

γ-Aminobutyric acid (GABA) and GABA-rich foods have shown anti-hypertensive and anti-depressant activities as the major functions in humans and animals. Hence, high GABA-producing lactic acid bacteria (LAB) could be used as functional starters for manufacturing novel fermented dairy foods. Glutamic acid decarboxylases (GADs) from LAB are highly conserved at the species level based on the phylogenetic tree of GADs from LAB. Moreover, two functionally distinct GADs and one intact gad operon were observed in all the completely sequenced Lactobacillus brevis strains suggesting its common capability to synthesize GABA. Difficulties and strategies for the manufacture of GABA-rich fermented dairy foods have been discussed and proposed, respectively. In addition, a genetic survey on the sequenced LAB strains demonstrated the absence of cell envelope proteinases in the majority of LAB including Lb. brevis, which diminishes their cell viabilities in milk environments due to their non-proteolytic nature. Thus, several strategies have been proposed to overcome the non-proteolytic nature of Lb. brevis in order to produce GABA-rich dairy foods.     

响应面法优化预酶解提取米渣蛋白的研究

为改善米渣中蛋白质的提取效果,采用中性蛋白酶酶解、碱溶两步法提取米蛋白.固定碱提取工艺,通过单因素实验,确定了预酶解的基本条件;并以pH和加酶量和反应时间为自变量,蛋白质提取率和质量分数为响应值,采用Box Behnken试验设计方法,对预酶解条件进行响应面（RSM）优化,确定了预酶解的最优工艺.最优条件下得到米渣蛋白质的提取率为78.27%,质量分数为72.86%,较直接碱提取,两者均有了一定的提高,且以蛋白质提取率提升幅度大,且优化条件下的实验结果与RSM回归方程预测值吻合良好.     

蛋白酶A对纯生啤酒泡持性的影响研究

该文研究蛋白酶A活力对纯生啤酒泡持性的影响,同时建立初始成品纯生啤酒及过滤前发酵液蛋白酶A活力限量值。结果表明,随着贮存时间的延长,纯生啤酒蛋白酶A活力和泡持值均呈下降趋势,最终残留蛋白酶A活力是影响其货架期泡持值的主要因素。纯生啤酒泡持值与蛋白酶A活力呈显著负相关（P＜0.01）。为了保证成品纯生啤酒在货架期内泡沫稳定性,发酵液出罐滤酒前的蛋白酶A活力应＜24×10-5 U/mL,相应成品纯生啤酒初始蛋白酶A活力应＜15×10-5 U/mL。该内控标准能为纯生啤酒生产企业控制泡持性提供指导。     

不同酶嫩化处理对牛肉物性的影响

本实验以牛肉为原料,利用TA-XT2i物性测定仪,研究了蛋白酶嫩化处理对牛肉及其制品物性的影响。结果表明:三种蛋白酶处理都能够较好地保持牛肉的持水力,复合蛋白酶、胰蛋白酶和木瓜蛋白酶处理后牛肉的持水力分别比对照牛肉高28.3%、11.1%和15.9%。另外,蛋白酶处理显著降低了牛肉的烹饪失水率、剪切力、咀嚼性、弹性、回复性、黏聚性等物性指标。其中,复合蛋白酶嫩化处理的效果优于胰蛋白酶及木瓜蛋白酶。显微照相表明酶作用后牛肉肌原蛋白被分解,肌原纤维断裂,肌纤维结构完整性的破坏可能是导致牛肉上述物性变化的主要原因。     

酶法改善卵白蛋白起泡性

采用碱性蛋白酶对卵白蛋白进行酶法改性处理以改善其起泡性。探讨酶解时间、底物质量分数、酶用量因素对卵白蛋白起泡性和泡沫稳定性的影响,在单因素试验基础上采用Box-Behnken中心组合试验设计优化酶法改性条件。结果表明,碱性蛋白酶酶法改性卵白蛋白改善其起泡性的最佳条件为：酶用量144 000 U/g、底物质量分数1.90%、酶解时间34 min。在此条件下酶法改性后卵白蛋白起泡性达202.0%,为未改性（120%）的1.683 倍。     

水解面筋蛋白高效制备谷氨酰胺结合肽的研究

采用食品级胃蛋白酶、胰蛋白酶、Protamex复合酶双酶组合对面筋蛋白进行水解制备Gln结合肽。先确定三种单酶各自对面筋蛋白适宜的初始pH、温度、酶浓度和底物浓度。并以双酶进行两两组合水解。在进行水解度控制实验时取样,分析水解液中氨基氮的含量,计算平均肽链长度,并用高效液相色谱分析水解液中Gln的得率。实验结果表明胰蛋白酶和胃蛋白酶联合水解的效果最好,条件为:胰蛋白酶在pH8.0,50℃,S%=12.8%(W/V),E%=9%(W/W)水解6h,再用胃蛋白酶pH2.0,40℃,E%=5%(W/W)条件下水解5h,可得到平均肽链长度为2.20个氨基酸残基的蛋白水解液,Gln得率最高,为60.43%。认为胰蛋白酶和胃蛋白酶组合是面筋蛋白中高效制备谷氨酰胺结合肽的最佳组合。     

Vascular Architecture Generates Fine Scale Variation in Systemic Induction of Proteinase Inhibitors in Tomato

Systemic induction following damage has been found in many plant species. Despite this widespread appreciation for the importance of induction, few studies have characterized the spatial variability of induction. We used tomato, Lycopersicon esculentum, to examine how damage to a single leaf affected the spatial distribution of systemic induction of proteinase inhibition in leaves above the damaged leaf. We crushed each leaflet of the second true leaf with forceps and measured the spatial distribution of proteinase inhibition in leaves 3, 4, and 5 at 8, 16, 24, 48, 72, and 120 hr. Constitutive levels of proteinase inhibitor activity were quantified in undamaged plants. We hypothesized that, due to vascular control of signal movement, systemic induction would show both among and within leaf variability. Following damage to leaf 2, induction was most pronounced in leaf 5 and minimal in leaf 3. In general, proteinase inhibitor activity was greatest at 24 hr and then declined. As predicted by vascular architecture, the near side of leaves in adjacent orthostichies showed higher induction than the far side of leaves. There was no increase in proteinase inhibitor activity in the undamaged neighboring plants. Overall our results demonstrate that systemic induction of proteinase inhibitors is partially controlled by vascular architecture and that future studies on systemic induction should examine the vascular architecture of the plants being studied. We argue that this spatial variation may influence the performance of herbivores sensitive to induced chemical changes.