耐冷菌的耐盐驯化与耐冷机制研究  

Isolation, Domestication and Character of Heterotrophic Psychrotrophs

作　　者：冯辉[1] 杨青 孙贻超[1] 徐海津[2] 

机构地区：[1]天津市环境保护技术开发中心,天津300191 [2]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2017年第4期28-34,共7页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：天津市自然科学基金重点项目(17JCZDJC32200)

摘　　要：为了改善生物降解法对低温高盐废水的处理效率,于冬季在环境中筛选获取耐低温菌PTJ3-3并通过驯化提高其盐耐受度.通过比较16Sr DNA序列结合Biolog菌种鉴定系统结果鉴定菌种为莓实假单胞菌(Pseudomanas fragi).参考国标法测定PTJ3-3在低温高盐条件下对有机物和总氮的降解效率,48 h其对有机物和总氮的降解率分别为10.34%和14.76%.由于高盐低温条件下微生物对低温的抗逆性与胞内的脂肪酸脱氢酶含量有关,进一步构建了报告基因融合质粒,检测PTJ3-3△-9脂肪酸脱氢酶的启动子强度.研究显示,PTJ3-3菌株△-9脂肪酸脱氢酶启动子序列与对照菌株A000152存在6处碱基差异,PTJ3-3的启动子启动报告基因表达的效率远超过对照菌株.实验结果对冬季高盐污水的生物降解处理具有应用价值,为研究低温微生物的耐冷机理提供了材料.Heterotrophic Psychrotrophs PTJ3-3 was isolated from the sewage and domesticated under high salinity condition.After identification,we investigated the degradation and promoter character related to cold tolerance gene.PTJ3-3 was identified by 16 Sr DNA and Biolog system.Degradation ability was tested by national standard method.Plasmids with reporter gene were constructed in order to study promoter efficiency.PTJ3-3 was identified as a Pseudomanas fragi.The degradation efficiency of COD and nitrogen was 10.34% and 14.76% respectively under 8 ℃,Na Cl 5%.Fatty acid desaturase was closely involeved in the ability of cold tolerance.The research on the△-9 fatty acid desaturase gene showed promoter showed that PTJ3-3 had higher promoter efficiency than control strains A000152.Our research suggested that PTJ3-3 provided important materials to investigate the mechanism of cold tolerance and could be applied effectively to biodegradate sewage in winter.

关 键 词：耐冷耐盐 菌种鉴定 降解 启动子强度 

分 类 号：Q939.9[生物学—微生物学]