培养条件对嗜水气单胞菌胞外蛋白酶合成与分泌的影响

对体外培养嗜水气单胞菌的胞外蛋白酶 (ECPase)产生条件进行了优化。结果发现 ,其产量与碳源、氮源、培养温度、培养基初始pH值等有关。蔗糖能促进蛋白酶的产生 ,NH4 + 抑制蛋白酶的产生。其最佳产酶条件为 :pH 7 5的0 0 2mol·L-1KC1,0 0 6mol·L-1K2 HPO4 ,5g·L-1的蔗糖和 5g·L-1的胰蛋白胨水 ,2 8℃ ,15 0r·min-1摇床培养 6 5h。在此条件下酶活性可达 2 0 8U·mL-1。     

旋毛虫成虫丝氨酸蛋白酶对S774A.1巨噬细胞的毒性作用

为探究旋毛虫成虫排泄分泌抗原(ES)中丝氨酸蛋白酶对宿主免疫调节作用,用PCR扩增出旋毛虫成虫期特异性丝氨酸蛋白酶基因Zh68,克隆至表达载体pET28a,转化到大肠埃希菌BL21( DE3),经诱导表达后的重组蛋白免疫接种动物获取抗血清.将抗体封闭前后的ES分别作用于S774A.1巨噬细胞,CCK-8检测巨噬细胞的增...     

寄生虫半胱氨酸蛋白酶研究进展

半胱氨酸蛋白酶(Cysteine protease,CP)是一类在酶的活性中心含有半胱氨酸残基的蛋白水解酶.CP属于木瓜蛋白酶家族,广泛分布于从病毒至脊椎动物的生物体中.对人体半胱氨酸蛋白酶的研究主要集中在细胞凋亡分子机制和恶性肿瘤的基因诊断与治疗等方面.而对寄生虫半胱氨酸蛋白酶研究的内容则更为广泛,其中包含半胱氨酸蛋...     

鸡肉蛋白酶水解工艺条件的研究

为探索鸡肉酶水解工艺条件及其动力学特性，分别采用木瓜蛋白酶、胃蛋白酶、中性蛋白酶、菠萝蛋白酶对鸡肉进行了水解试验，得出最优酶为木瓜蛋白酶；由此选用木瓜蛋白酶做正交试验，考察水解温度、时间、加酶量、pH值和固液比5个因素对酶水解的影响，并确定出最优水解条件为温度50℃，时间7 h，加酶量2.4%(以鸡肉的质量百分数计)，pH 7.0，固液比1∶4；在此条件下，水解度可达26.07%。在此基础上由试验数据推导出描述木瓜蛋白酶水解鸡肉的动力学方程，可为鸡肉酶解生化反应器的设计和鸡肉水解蛋白液或蛋白粉的开发提供一定的理论依据。     

Evidence of peptides in low-molecular-weight fractions of soil organic matter

Summary Organic matter was extracted from three soils, a Berwick sandy loam, a Franklin loamy sand, and a Cumberland silty loam. The extracts were separated into high (>8000 daltons) and low-molecular-weight (<8000 daltons) fractions using gel filtration. Reverse-phase high performance liquid chromatography at 214 nm was used to separate the peptides into low-molecular-weight fractions. Peptide samples were collected with an integrated fraction collector and hydolyzed with an immobilized protease column reactor. High performance liquid chromatography with fluorescence detection was used to determine the amino-acid contents of the collected samples. The results indicated that peptide intermediates are present in soil size fractions. Greater quantities of several amino acids were released from the peptide hydrolyzates of the Berwick sandy loam and Franklin loamy sand, compared with the Cumberland silty loam, an uncultivated soil. These findings indicate that organic intermediates (e.g., peptides) are more prevalent in biologically active soils than in relatively inert soils.     

甘蓝黑腐病菌rPf基因簇对胞外蛋白酶Ⅰ基因表达的调控

甘蓝黑腐病黄单胞菌（ＸａｎｔｈｏｍｏｎａｓｃａｍｐｅｓｔｒｉｓＰｖ．ｃａｍｐｅｓｔｒｉｓ）产生的胞外蛋白酶Ⅰ在致病的早期阶段起重要作用，该酶以及其它胞外酶和胞外多糖的合成受一致病因子调控基因簇（ｒＰｆ基因簇）的正向调控。本研究利用带有β－半乳糖苷酶报道基因（ｌａｃＺ）的转座子Ｔｎ５－Ｂ２０诱变蛋白酶Ⅰ基因克隆，获得了ｌａｃＺ在蛋白酶Ⅰ基因启动子控制下表达的Ｔｎ５－Ｂ２０插入突变质粒。通过将这种突变质粒导入野生型和各ｒｐｆ基因突变体菌株后，测定ｌａｃＺ基因在细胞生长周期中的表达水平，不仅进一步证实了这些ｒｐｆ基因对蛋白酶Ⅰ基因的正向调控作用，而且明确了它们的调控水平．发现ｒｐｆＡ、ｒｐｆＣ、ｒｐｆＥ、ｒｐｆＧ或ｒｐｆＨ突变后，蛋白酶Ⅰ基因的转录会降低９０％左右，而ｒｐｆＢ突变后，蛋白酶Ⅰ基因的转录只降低４８％。     

Colostral Trypsin-Inhibitor Capacity in Different Animal Species

Colostral trypsin-inhibitor capacity was monitored during the first two weeks from parturition. The colostrum of the mare, sow, cow and ewe showed high antitrypsin activity at parturition, decreasing to about one hundredth during the first week. Canine milk remained on a relatively high antitrypsin level, and human milk was poor in antitrypsin from childbirth. The antitrypsin content seems to parallel the known changes in the colostral immunoglobulin levels of different species. The role of antitrypsin in protection of immunoglobulins from proteolytic damage during passive transfer of immunity to the newborn is obvious.     

Protein degradation and protease activity of chum salmon (Oncorhynchus keta) muscle during spawning migration

Sarcoplasmic protein content decreased significantly and myosin heavy chain was degraded gradually during spawning migration of chum salmon (Oncorhynchus keta). Acid and neutral proteinase activities increased significantly during spawning migration. These proteinase activities were higher in females than in males. High levels of acid proteinase were not caused by the injury of lysosomal membranes. Considering the physiological states of chum salmon, neutral proteinase activity might be related to the degradation of muscle protein. As the changes in serum sex steroids were similar to those in the protease activities during spawning migration, it was considered that high levels of protease activity during spawning migration were closely related to the serum levels of androgens.     

Co~（2＋）对渗透胁迫下小麦幼苗叶片乙烯产生率及蛋白酶活性的影响

在－１．０ＭＰａＰＥＧ渗透胁迫下，小麦幼苗叶片乙烯产生速率和蛋白酶活性随胁迫时间延长递增，蛋白质和叶绿素含量递减。在－１．０ＭＰａＰＥＧ溶液中加入２ｍｍｏｌ／ＬＣｏＣｌ２处理可以抑制乙烯的产生及蛋白酶的活性，对蛋白质和叶绿素起到保护作用。同时还观察到，渗透胁迫过程中乙烯的产生速率与叶片的衰老过程具有正相关性，加入Ｃｏ２＋能一定程度的减轻渗透胁迫造成的伤害。     

枯草芽孢杆菌发酵鱼粉产蛋白酶及其酶解效果

为开发以鱼粉为原料的优质饲料蛋白源以及相关保健食品,开展了用不同鱼粉为氮源发酵枯草芽孢杆菌生产蛋白酶及其酶解效果的研究,比较了不同菌种的发酵效果,确定了最佳发酵时间.通过蛋白酶活性测定和氨基态氮含量测定分析了不同菌种的产酶能力和酶活性大小及其酶解效果.结果表明,用罗非鱼鱼粉和处理过的鳕鱼鱼粉发酵商业化的工业菌种枯草芽孢杆菌产生的酶活性显著高于鲢鱼鱼粉（P〈0.05）,发酵后上清液中的氨基态氮含量也显著高于鲢鱼鱼粉（P〈0.05）,即工业化的枯草芽孢杆菌对不同鱼粉的发酵效果不同,罗非鱼鱼粉和处理过的鳕鱼鱼粉发酵效果显著优于鲢鱼鱼粉.