Digestive Proteases of Pacu,Piaractus mesopotamicus,Fed on Distinct Protein-Starch Diets

Abstract

Fish adjust the enzyme profile to dietary protein and carbohydrate. A balance of nutrients optimizes the enzyme profile and metabolic performance offish. Dietary levels (10, 20, 30 and 40%) were combined to four levels of starch (57, 32, 21 and 4%) to feed juvenile pacu, Piaractus mesopotamicus. Acid and alkaline proteases were color-imetrically determined by hemoglobin and azocasein hydrolysis, respectively. Liver, white and red muscle glycogen, glucose and free amino acids were colorimetrically quantified. Those enzyme activities were induced by dietary protein. Enzyme responses to the feeding and the metabolic profile of the tissues suggest the best protein/starch ratio as 30%/21%. The highest levels of protein (30%-40%) impaired nutrient uptake.     

蛋白酶和木聚糖酶对肉仔鸡生长性能、血液指标和消化机能的影响

以蛋白酶和木聚糖酶分别作为消化酶(或内源酶)和非消化酶(或外源酶)的代表,研究小麦型日粮中添加不同水平蛋白酶和木聚糖酶对1~21日龄肉仔鸡生长性能、血液指标、消化器官指数及肠道组织学形态的影响,为酶制剂在家禽日粮中的合理使用提供理论依据。1日龄健康肉仔鸡972只,随机分为9个处理,每处理9个重复,每重复12只,采用双因子完全随机试验设计,分别饲喂在基础日粮中添加不同水平蛋白酶(0、4 000、32 000 U/kg)和木聚糖酶(0、1 000、8 000 U/kg)的试验日粮,试验期21 d。结果表明,4 000 U/kg蛋白酶+1 000 U/kg木聚糖酶组显著降低肉仔鸡采食量(P<0.05),32 000 U/kg蛋白酶添加水平趋于提高采食量(P=0.080),但对饲料转化率无显著影响(P>0.05)。32 000 U/kg蛋白酶添加水平显著降低血糖含量(P<0.05)。1 000 U/kg和8 000 U/kg木聚糖酶水平均趋于提高回肠指数(P=0.051)。32 000 U/kg蛋白酶组趋于降低空肠绒毛高度(P=0.081)。与8 000 U/kg木聚糖酶组相比,32 000 U/kg蛋白酶组显著增加...     

蛋白酶高产菌株的紫外诱变选育

［目的］提高地衣芽孢杆菌B7的产蛋白酶能力。［方法］采用福林酚法测定酶活力,通过诱变育种试验和遗传稳定性试验选育出遗传性能稳定的蛋白酶高产菌株。［结果］紫外线照射时间为2 min,菌落的总致死率为62.47%。371株菌株的HC值变化小于5%,53株菌株的HC值正变大于5%,76株菌株的HC值负变大于5%。正变大于10%的12株菌株的HC值是出发菌株的1.18-1.34倍。所有变异株的蛋白酶活力均显著高于出发菌株,其中菌株B720和B749的蛋白酶活力达50 U/ml,比出发菌株提高了40%以上。经过复筛得到菌株B7 49,诱变前后其最高酶活分别为33.97和50.29 U/ml。遗传稳定性试验中菌株B749的蛋白酶活力变化小于10%。［结论］该试验成功选育出的遗传性能稳定的蛋白酶高产菌株B749可用于发酵法制备大豆肽。     

羊骨降血压肽制备工艺的研究

试验以羊骨为原料,采用酶技术,优选降血压活性最高的酶及其最佳酶解条件。结果表明:中性蛋白酶水解羊骨所得的ACE抑制率最高,达66.0%,其酶解条件为:酶浓度为100 U·g-1、底物浓度为25%、pH值7.0、温度为50℃、时间6 h。     

昆虫杆状病毒表达系统中重组蛋白质的降解及其对策

昆虫杆状病毒基因表达系统是目前比较高效的真核表达系统之一,但重组蛋白质的降解成为影响该系统生产效率的一个重要问题。就该系统中蛋白质降解的相关问题及其对应策略进行了探讨。     

Regulation of extracellular protease activity in soil in response to different sources and concentrations of nitrogen and carbon

A large proportion of the nitrogen (N) in soil is in the form of proteinaceous material. Its breakdown requires the activity of extracellular proteases and other decomposing enzymes. The goal of our study was to better understand how carbon (C) and N availability affect soil protease activity. Several aerobic incubations were carried out with ammonium (NH₄⁺) and proteins as N sources and cellulose as the main C source. A strong increase in protease activity was observed when proteins were added, the increase depending on the amount of protein added and its solubility. Protease synthesis was clearly substrate induced, as NH₄⁺ had no effect. During this substrate induced phase, the addition of glucose but not NH₄⁺ resulted in protease repression, indicating that the level of protease synthesis was determined by the need for C rather than N. After 1 month of incubation, protease activity remained relatively constant over time and was closely related to microbial biomass N. Different concentrations of mineral N in soil solution had no direct effect on protease activity. However, during this stationary phase, protease activity could be repressed by glucose and NH₄⁺ in a treatment with low mineral N content while in treatments with a higher N availability no repression was observed. We hypothesize that the need for N determined protease activity in the treatment with limited N availability. The addition of NH₄⁺ allowed for reallocation of C and N away from protease synthesis, leading to the observed decrease in protease activity. The repression by glucose may be attributed to shifts in the pathway of microbial NH₄⁺ assimilation. The results emphasize the close links between the microbially mediated cycles of organic C and N.     

长牡蛎中I84蛋白酶抑制因子家族的扩张和功能分化

蛋白酶抑制因子是极其多样的蛋白质或多肽,能抑制蛋白酶的水解活性。研究发现蛋白酶抑制因子能通过抑制病原蛋白酶活性,从而抑制病原的入侵。I84蛋白酶抑制因子家族是MEROPS数据库中新增的一个家族,其部分成员在免疫防御过程中的作用得到了一定的揭示。为探究I84蛋白酶抑制因子家族在长牡蛎中的分布和功能状况,实验鉴定了长牡蛎中23个潜在的I84家族基因,根据系统进化树分析,挑选了5个同源基因进行时空表达和功能分析。首先,利用克隆技术验证了长牡蛎中5个I84家族同源基因CgSi3、CgSi5、CgSi6、CgSi16和CgSi19可表达性。结果显示,5个基因在外套膜、闭壳肌、性腺、血淋巴细胞、肝胰腺和鳃等6个组织中均表达,但肝胰腺中表达量显著高于其他组织。同时,5个基因在长牡蛎幼体不同发育阶段表达模式不同,其中各基因在受精卵时期均不表达,CgSi3表达量则在眼点幼虫期显著上升后下降,而CgSi6在壳顶幼虫期开始表达后,表达量随长牡蛎发育而持续增加。另外,对牡蛎进行病原相关模式分子(PAMPs)注射后,5个基因也表现出不同的表达模式,其中LPS和PGN注射后CgSi6表达量变化明显,而poly (...     

黑龙江几个大豆品种中抗营养因子含量的分析

本研究测定了１３个黑龙江大豆品种中的抗营养因子含量,包括胰蛋白酶抑制因子,糜蛋白酶抑制因子,凝集素。发现高蛋白含量的“东农４２”品种抗因子含量最低,ＴＩＡ为３６．７２ｍｇ／ｇ,ＣＩＡ为７．９８ｍｇ／ｇ,凝集素为１．９７ｍｇ／ｇ。脂用型品种“７１４３５”的蛋白质含量低,抗因子成分的含量却是最高,ＴＩＡ６２．２５ｍｇ／ｇ,ＣＩＡ３５．６１ｍｇ／ｇ,凝集素为３．８６ｍｇ／ｇ。     

投饲蚕豆对异育银鲫生长、肉质及肠道蛋白酶活力的影响

分别以配合饲料、浸泡蚕豆和发芽蚕豆饲喂平均体重23.7 g的异育鲫鱼(Allogynogenetic crucian carp),观察对其生长性能、肌肉品质和前肠蛋白酶活力的影响。经44 d养殖实验后,配合饲料组,浸泡蚕豆组和发芽蚕豆组的鱼体增重率分别为95.64%、33.54%和42.62%,饵料系数为1.97、5.09和4.15,投饲蚕豆显著降低了异育银鲫的增重率,提高了饵料系数(P<0.01);对肉质的分析表明,浸泡蚕豆组和发芽蚕豆组的肌肉失水率,肌原纤维耐折力和肌纤维直径显著提高(P<0.01),肌肉粗脂肪含量由1.92%(配合饲料组)降至1.05%和1.14%(P<0.01)。对肠道蛋白酶活力的分析表明,投饲蚕豆显著降低了前肠蛋白酶的活性(P<0.01)。上述研究表明,饲喂蚕豆可改善异育银鲫的肉质,但对其生长有抑制作用。     

温度对鲫血液生化指标和消化酶的影响

采用室内模拟方法,研究了鲫(Carassius auratus)在9℃、16℃、25℃、30℃温度条件下的血液学指标和消化酶活力的变化。结果显示,鲫血清无机成分中的镁、磷在16℃时存在最大值,而其它成分变化不显著。在9～25℃水温范围内,甘油三脂、总胆固醇、总蛋白、白蛋白、球蛋白等均随水温上升而上升,并在25℃时达到最大值,而超过25℃后,甘油三脂、总胆固醇、总蛋白、白蛋白、球蛋白等含量均随水温升高而下降。在9～30℃水温范围内,血清中谷丙转氨酶和谷草转氨酶活性随水温上升先升高、后下降,在25℃时达到最大值。鲫肠道中,蛋白酶和脂肪酶活性在9～25℃阶段升高,25～30℃时下降,蛋白酶和脂肪酶的活性均在25℃时最高;而肝胰脏中蛋白酶和脂肪酶活性随温度变化不明显。