家蚕蛾吐出液的酶活性及蛋白质种类鉴定  

Enzymatic Activity and Protein Species Identification of Spit Liquid from Bombyx mori Moths

作　　者：祝元锋[1,2] 王赵群 何宁佳[1] 冯丽春[1] 

机构地区：[1]西南大学生物技术学院,家蚕基因组生物学国家重点实验室,重庆400716 [2]第三军医大学第一附属医院中心实验室,重庆400700

出　　处：《蚕业科学》2014年第3期452-457,共6页ACTA SERICOLOGICA SINICA

基　　金：现代农业产业技术体系建设专项(No.CARS-22)

摘　　要：家蚕羽化时用口腔吐出液溶解前端茧层的丝胶及少量丝素后脱茧而出,测定吐出液中各种酶类的活性以及鉴定吐出液中的蛋白质种类,对解析家蚕羽化过程的溶茧机制具有重要意义。在家蚕羽化前解剖收集嗉囊液,在家蚕羽化时收集蛾吐出液,解剖获取蛾下颚晶体,测定3组样品中的蛋白酶、酯酶、纤溶酶和丝胶酶活性:在蛾吐出液和嗉囊液中皆检测到4种酶的活性,2组检测样品除蛋白酶活性接近外,酯酶、纤溶酶和丝胶酶活性则是蛾吐出液比嗉囊液分别高出21%、57%、26%;下颚晶体不具有纤溶酶活性,其他3种酶的活性也皆比蛾吐出液和嗉囊液低。采用液相色谱-串联质谱(LC-MS/MS)分析技术在家蚕蛾吐出液中鉴定出29种蛋白质,其中包括14种酶类、4种蛋白酶抑制剂、5种30K蛋白、1种转运相关蛋白、2种免疫相关蛋白和3种其他蛋白质。研究结果显示了蚕蛾吐出液成分的复杂性,同时说明家蚕羽化时吐出液对丝胶和部分丝素的水解除了溶茧酶的作用外,可能还有其他成分的参与。Silkworm （Bombyx mori） moth dissolves apex of cocoon layer sericin and a small amount of fibroin by spit liquid at the stage of eclosion. It is of great significance to measure activity of various enzymes and identify protein species in moth spit liquid for understanding molecular mechanisms of cocoon dissolution during the ecIosion process. The craw liquid was dissected and collected before the eclosion, while spit liquid and maxilla crystal of silkworm moths were collected and dissected at the stage of eclosion. The activities of protease, esterase, plasmin, and sericinase in three samples were measured. The results showed that both spit liquid and craw liquid had the above four enzymes. In these two samples, ac- tivity of protease was close while that of esterase, plasmin, and sericinase in spit liquid was 21%, 57%, and 26% higher than that in craw liquid respectively. Maxilla crystal did not have plasmin activity and had lower activity of the other three en- zymes than spit liquid and craw liquid. Using LC-MS/MS analysis technology, we identified 29 proteins in the spit liquid in- cluding fourteen proteases, four protease inhibitors, five 30K proteins, one transporter-related protein, two immune-related proteins, and three other proteins. Our results revealed the complexity of moth spit liquid, and indicated that there are othercomponents involved in the hydrolysis of sericin and fibroin at the eclosion stage apart from cocoonase.

关 键 词：家蚕 羽化 吐出液 嗉囊液 下颚晶体 酶活性 蛋白质 液相色谱一串联质谱 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q55[农业科学—畜牧兽医]