环境中甾体激素降解菌的筛选及功能基因的克隆  

Isolation of A New Steroid Degrading Bacterial Strain and Functional Gene Cloning

作　　者：于源华[1] 宋禹[1] 何秀霞[1] 王宝德[1] 刘华 李金海[1] 熊光明[1] 

机构地区：[1]长春理工大学生命科学技术学院,长春130022 [2]长春卫尔赛生物药业有限公司,长春130022

出　　处：《长春理工大学学报（自然科学版）》2010年第1期108-111,共4页Journal of Changchun University of Science and Technology(Natural Science Edition)

基　　金：吉林省科技厅重大支撑项目(20096012)

摘　　要：利用生物技术解决环境中日益增长的甾体类激素污染的环境问题,寻找相关的菌及其关键基因就非常重要。本研究利用含1.6-4.1%NaCl的SIN培养基从大连海港附近所取海水样中筛选能够降解并利用甾体激素作为唯一碳源和能源的微生物。ELISA检测3-羟基类固醇脱氢酶/羰基还原酶(3-HSD/CR)活性。利用PCR技术克隆3,17-羟基类固醇脱氢酶基因,ELISA技术检测该基因的活性。结果表明筛选到的微生物为一种革兰氏阴性菌,命名为H5-1,具有3-HSD/CR活性,对PCR克隆的3,17-羟基类固醇脱氢酶基因经测序比对同源性为99%,ELISA检测初步证实了该基因的活性。该基因的成功克隆为后期转入受体细胞进行目的蛋白的表达、酶活性鉴定及构建甾体激素降解工程菌株等工作奠定基础。Biological technology solving the growing environment of steroid hormone contamination of environmental problems,finding related bacteria and its genes of key enzyme are very important. In this work,we have isolated a new bacterium （strain H5-1）from the Bohai Sea at Dalian,China,Which is able to degrade steroids like testosterone,estradiol and cholesterol.Strain H5-1was characterized as being a gram-negative bacterium that could be best grown in SIN medium supplemented with 1.6-4.1%NaCl. By PCR and ELISA testing,confirmed the H5-1 with activity of 3 -Hydroxysteroid dehydrogenase/ carbonyl reductase （3 -HSD/CR）.Besides,we found that the H5-1 has another kind of steroid hormone-degrading enzyme, 3,17 -Hydroxysteroid dehydrogenase （3,17 HSD）. These studies provide a basis for the latter parts of the research.

关 键 词：甾体激素 H5-1菌株 生物降解 

分 类 号：S937[农业科学—渔业资源]