鸭肝蛋白酶的分离纯化及其部分性质研究   总被引：1，自引：1，他引：1  

岑亮  张丽丽  邱慧  郭小路  唐云明 《西南大学学报(自然科学版)》2007,29(2):97-101

经Tris-HCl缓冲液抽提、硫酸铵分级沉淀、DEAE-Sepharose Fast Flow离子交换层析、Sephacryl S-200凝胶过滤层析,得到鸭肝蛋白酶电泳纯制品．纯化倍数为650.16倍,回收率为19.23%．经SDS-PAGE和凝胶过滤测得该酶的分子量为35kD．该酶在pH6.0～11.0,温度低于25℃稳定,当温度上升到55℃时活性迅速下降．该酶在pH10和60℃时达到最高活性．进一步研究得知：Cu^2＋离子对该酶有明显促进作用,而Mn^2＋却使酶几乎失活．以酪蛋白为底物,在pH10.0、37℃时测得米氏常数Km值为1.33%．  相似文献   

水相酶法提取蛋黄油工艺的响应面分析法研究   总被引：6，自引：0，他引：6  

刘杰  徐明生  刘建涛  赵利  万细妹 《江西农业大学学报》2007,29(5):821-826

采用蛋白酶水解鲜鸡蛋蛋黄提取蛋黄油,通过响应曲面分析法建立蛋黄油提取率的二次多项数学模型,并考察酶添加比例、底物质量浓度和反应时间对蛋黄油提取率的影响,优化出蛋黄油提取率达到56.81%的工艺参数:酶添加比例为2.04%,底物质量浓度为20.28%,反应时间为14 h。  相似文献   

旋毛虫成虫丝氨酸蛋白酶对S774A.1巨噬细胞的毒性作用     

于建立  白雪  刘国兴  吴秀萍  王学林  刘明远 《动物医学进展》2011,(10):5-9

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...  相似文献   

寄生虫半胱氨酸蛋白酶研究进展   总被引：1，自引：0，他引：1  

宋军科  才学鹏  骆学农  林青  窦永喜  于三科 《动物医学进展》2011,(12):95-98

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...  相似文献   

鸡肉蛋白酶水解工艺条件的研究   总被引：11，自引：2，他引：11  

谢永洪  刘学文  王文贤  冉旭 《农业工程学报》2004,20(5):207-210

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。  相似文献   

Evidence of peptides in low-molecular-weight fractions of soil organic matter   总被引：2，自引：0，他引：2  

P. R. Warman  R. A. Isnor 《Biology and Fertility of Soils》1989,8(1):25-28

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.  相似文献   

Colostral Trypsin-Inhibitor Capacity in Different Animal Species     

M. Sandholm  T. Honkanen-Buzalski 《Acta veterinaria Scandinavica》1979,20(4):469

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.  相似文献   

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

Ando  Seiichi  Hatano  Mutsuo  Zama  Kōichi 《Fish physiology and biochemistry》1986,1(1):17-26

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.  相似文献   

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   总被引：2，自引：0，他引：2  

李朝周 《甘肃农业大学学报》1996,31(4):376-379

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。  相似文献   

桔小实蝇丝氨酸蛋白酶基因（BdorSer）的克隆与表达分析     

胡黎明  曾玲  申建梅 《西北农林科技大学学报(自然科学版)》2012,40(7):70-76

【目的】克隆桔小实蝇丝氨酸蛋白酶基因（Bactrocera dorsalis serine protease,BdorSer）,研究BdorSer在不同组织和不同发育时期的表达情况,探索其可能参与的生理过程。【方法】采用反转录聚合酶链式反应(RT-PCR)和快速扩增cDNA末端(RACE)技术克隆BdorSer,对其编码蛋白氨基酸序列信息和进化关系进行分析;采用实时荧光定量PCR（real-time PCR）方法,研究BdorSer mRNA在桔小实蝇不同组织及不同发育时期的相对表达量。【结果】克隆获得了BdorSer,该基因ORF全长1 239 bp。氨基酸序列一致性分析表明,BdorSer与拟暗果蝇（Drosophila pseudoobscura）丝氨酸蛋白酶（登录号XP_001352960）氨基酸序列的一致性最高,为65.8%。实时荧光定量PCR分析表明,BdorSer mRNA在桔小实蝇雌雄虫的不同组织中都有表达,且在触角中的表达量最高,分别是雌虫胸部的43.6和26.8倍。BdorSer mRNA在生殖节中表现出较大的性别差异表达特征,雄虫生殖节中的mRNA表达量是雌虫的18.25倍。BdorSer mRNA几乎在昆虫发育的各个时期都有表达,其中在卵及1,2,3龄幼虫中的表达量分别是10 d 蛹的 0.68,1.63,4.31和15.8倍。BdorSer mRNA随蛹的发育其表达量逐渐减低,1,4,7 d蛹中的表达量分别是10 d蛹的325,125和41倍。【结论】克隆获得了BdorSer,其表达的BdorSer蛋白可能在雄虫的生殖生理中发挥了作用,同时也可能参与了桔小实蝇的变态发育过程,尤其是1 d蛹的发育过程。  相似文献