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。     

肉桂色曲霉9824饲用酸性蛋白酶的产生、性质及应用研究

本文对肉桂色曲霉9824菌株酸性蛋白酶产酶条件，酶学性质和在断奶仔猪的喂养效果进行了研究。结果表明，采用麸皮与豆饼比例6：4，氯化铵浓度1．5％，物料加水比1：1，菌株接种量0．3％，经66h发酵，其产酶活较高，该酶最适酸碱度pH3.0，最适作用温度为40℃，用pH4．0的乳酸缓冲液在50℃下提酶较完全。该酶具有较好的热稳定性，在100℃，1h可保持80％的酶活力。小鼠急性毒性试验证明该酶安全无毒。以0．1％添加到当地饲喂秧藤粉的断奶仔猪日粮中，经40d喂养，试验组比对照组日增重和饲料转化率分别提高13．2％和2．4％，并有效地控制了腹泻。     

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.     

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.     

变性豆粕中蛋白质的酶水解特性的研究

变性豆粕是由大豆浸油后,经高温脱溶所得。本文以高温变性脱脂大豆粕为原料,用正交实验法对变性后在蛋白酶作用下的水解特性进行了深入研究。选用国产1398中性蛋白酶为水解酶对变性豆粕进行水解,研究了变性豆粕中蛋白质溶出率随温度、pH值、时间、底物浓度及用酶量的变化规律,找到了水解变性豆粕的最佳实验条件。为生产实践提供了基础数据,该研究结果对其它蛋白质原料的水解特性研究也具有参考价值。研究结果表明：1398中性蛋白酶水解高温变性豆粕的最佳条件为：温度45℃,时间3h,底物浓度1．0％,用酶量1600u/g,pH值7．0,在此条件下,变性豆粕中蛋白质可有90．71％水解溶出。     

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.     

Biochemical properties of two isoforms of trypsin purified from the Intestine of skipjack tuna (Katsuwonus pelamis)

Two trypsins (A and B) from the intestine of skipjack tuna (Katsuwonus pelamis) were purified by Sephacryl S-200, Sephadex G-50 and DEAE-cellulose with a 177- and 257-fold increase in specific activity and 23% and 21% recovery for trypsin A and B, respectively. Purified trypsins revealed a single band on native-PAGE. The molecular weights of both trypsins were 24 kDa as estimated by size exclusion chromatography and SDS–PAGE. Trypsin A and B exhibited the maximal activity at 55 °C and 60 °C, respectively, and had the same optimal pH at 9.0. Both trypsins were stable up to 50 °C and in the pH range from 6.0 to 11.0. Both trypsin A and B were stabilised by calcium ion. Activity of both trypsins continuously decreased with increasing NaCl concentration (0–30%) and were inhibited by the specific trypsin inhibitors – soybean trypsin inhibitor and N-p-tosyl-l-lysine chloromethyl ketone. Apparent K_m and K_cat of trypsin A and B were 0.22–0.31 mM and 69.5–82.5 S⁻¹, respectively. The N-terminal amino acid sequences of the first 20 amino acids of trypsin A and B were IVGGYECQAHSQPPQVSLNA and IVGGYECQAHSQPPQVSLNS, respectively.     

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。     

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。     

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.