乌鳢源维氏气单胞菌胞外产物的生物学活性分析

Investigation on the enzymatic activity, pathogenicity and protein components of extracellular products of Aeromonas veronii strain isolated from Channa argus

关键词:  维氏气单胞菌  胞外产物  酶活  LC-MSMS  GO分类分析

Key words:  Aeromonas veronii  extracellular products  enzyme activity  LC-MSMS  GO classification analysis

摘要:

为了研究维氏气单胞菌胞外产物在乌鳢感染中的致病作用以及维氏气单胞菌的致病机理，本试验以乌鳢源维氏气单胞菌WL161为研究对象，提取其胞外产物，采用打孔法测定其胞外产物的酶活性，并对与毒力密切相关的溶血活性进行进一步的溶血谱研究，同时分析其致病性。应用LC-MSMS方法对其胞外产物蛋白成分进行鉴定，利用Gene Ontology(GO)对已鉴定蛋白进行生物学过程、分子功能和细胞组分的分类分析。结果表明：WL161菌株具有淀粉酶活性、脂肪酶活性、蛋白酶活性和溶血活性，不具有明胶酶活性；研究发现ECP对其他多种动物红细胞均有溶血活性，对鱼类红细胞溶血性较强，但对鸡红细胞无溶血活性。其胞外产物共检测出118种蛋白，共参与40种生物学过程，主要涉及碳水化合物代谢过程、几丁质分解代谢过程、DNA结合等；69种分子功能，主要涉及ATP结合、金属离子结合、碳水化合物结合等；19种细胞组分，主要包括胞外区、细胞质、细胞外膜等。

Abstract:

In order to study the pathogenic role of extracellular products(ECP) and pathogenic mechanism of Aeromonas?veronii.This study extracted the extracellular products from Aeromonas?veronii WL161.The enzyme activity and hemolytic activity of extracellular products were determined by punch method.LC-MSMS and Gene Ontology method were used to identify the protein components of the extracellular products and analysed biological process,molecular function and cellular components,respectively.The results?showed?that WL161 had amylase and lipase activity, protease and hemolytic activity,without gelatinase activity.ECP of most animals have a hemolytic activity, and the hemolytic effect of ECP showed a high hemolytic activity to fish erythrocyte, while no hemolytic activity to chicken erythrocyte.118 kinds of proteins were detected by the extracellular products.These proteins involved in 40 kinds of biological processes,especially carbohydrate metabolism process,chitin catabolic process and DNA binding;69 kinds of molecular functions,mainly involved ATP binding,metal ion binding,carbohydrate binding;19 kinds of celelular components, including extracellular region,cytoplasm and cell outer membrane.