一株Chitinophaga菌株的鉴定、优化及其活性蛋白的初步纯化  

Identification,Optimization and Preliminary Purification of Active Protein of Chitinophaga Strain

作　　者：刘冰花[1] 杨林[1] 罗玲[1] 蒲丽娟[1] 褚应文 杨港[1] LIU Binghua YANG Lin LUO Ling PU Lijuan CHU Yingwen YANG Gang(School of Medicine, Chengdu University, Chengdu 610106, China)

机构地区：[1]成都大学医学院(护理学院),四川成都610106

出　　处：《成都大学学报（自然科学版）》2017年第1期1-6,共6页Journal of Chengdu University（Natural Science Edition）

基　　金：四川省教育厅自然科学重点课题(12ZA207)资助项目

摘　　要：为确定前期获得的可产生抗血栓物质细菌的分类地位、优化发酵条件及对活性物质初步纯化.利用形态学特征、理化性质、脂肪酸含量、G+C含量、16S rRNA序列同源性及DNA-DNA杂交率等方法鉴定菌株MJM38,利用纤维蛋白平板法检测溶栓活性,利用纤维蛋白原平板法检测抗凝活性,利用单因素法筛选菌株MJM 38产生抗血栓活性物质的最佳培养条件,利用盐析、超滤及分子筛层析等方法对其活性物质进行了初步分离.结果发现,菌株MJM 38属于Chitinophaga菌,初步确定抗血栓活性物质为蛋白质,其分子量大小在30 k Da^100k Da之间,菌株MJM 38在培养时间为4 d、培养温度为25℃~30℃、发酵培养基初始p H值为7时产生的抗血栓物质的溶栓活性最大.The paper is going to determine the taxonomic status of the strain which can produce antithrom- botic material, optimize the fermentation conditions and preliminarily purify the active substance. The strain MJM 38 is identified by morphological characteristics, physical and chemical properties, fatty acid content, G ＋ C content, 16S rRNA sequence homology, phylogenetic tree and DNA-DNA hybridization rate. The fi- brinolysis activity is detected by the fibrin plate and the anticoagulant activity is detected by fibrinogen plate. The optimum culture condition for MJM 38 to produce antithrombotic substance is selected by single- factor method. The initial prefracfionation of the active substance is done briefly by salting out, hyperfiltra- tion and molecular sieve chromatography, etc. The results show that the strain MJM 38 belongs to Chiti- nophaga. The antithrombotic activity substance is initially identified as protein and its molecular weight is between 30- 100 kDa. It is also found that when the optimum culture conditions for strain MJM38 to pro- duce antithrombotic protein are 4 d, 25 - 30 ℃, and pH 7, the fibrinolysis activity of the antithrombotic substance achieves the maximum.

关 键 词：Chitinophaga 鉴定 优化 纯化 抗血栓 蛋白 

分 类 号：Q939.124[生物学—微生物学]