一株芘降解菌的分离鉴定及其降解效果  

Isolation,identification,and degrading effect of a pyrene-degrading strain SE12

作　　者：张巧巧[1] 赵叶君[1] 杨超光[1] 刘奋武[1] 何健[1] 沈标[1] 冉炜[1] 

机构地区：[1]南京农业大学固体有机废弃物资源化高技术研究重点实验室,南京210095

出　　处：《应用生态学报》2010年第7期1851-1858,共8页Chinese Journal of Applied Ecology

基　　金：国家自然科学基金项目(20677027)资助

摘　　要：以芘为唯一碳源,采用平板升华法,从徐州市卧牛山焦化厂周围污染土壤中分离得到一株芘降解菌SE12.经形态观察、生理生化试验和16S rDNA鉴定,该菌株属于分枝杆菌属(Mycobacterium sp.)菌株,与快速生长型非致病性南非分枝杆菌(M.austroa fricanum ATCC33464)的同源性达到98%.SE12降解芘的最适pH和温度为pH9和30℃.当土壤芘初始含量为100和200mg.kg-1,SE12接种量为107CFU.g-1时,30℃培养28d后土壤芘降解率分别达到97%和99%.利用双加氧酶基因的同源序列引物nidAF/nidAR和nidBF/nidBR进行扩增,得出了该菌株编码双加氧酶大亚基和小亚基的基因片段,它们与已知降解芘的分枝杆菌的双加氧酶基因具有高度同源性.Using pyrene as the sole carbon and energy source,and by the method of plate sublimation,a strain SE12 was isolated from a contaminated soil near Woniushan Coking Plant in Xuzhou,China.According to the morphological,biochemical,and 16S rDNA analyses,this strain was identified as Mycobacterium sp.,with 98% of homology to the rapid-growth nonpathogenic strain M.austroafricanum ATCC 33464.The optimum pH and temperature for the degradation of pyrene by SE12 were pH 9 and 30 ℃.When the soil samples were added with 100 and 200 mg·kg-1 of pyrene and inoculated with 107 CFU·g-1 of SE12,the degradation rates of pyrene reached to 97% and 99%,respectively after 28 days incubation at 30 ℃.By using primer-pairs nidAF/nidAR and nidBF/nidBR for amplification of ring-hydroxylating dioxygenase genes,it was shown that SE12 had the fragments of encoded large and small subunits of dioxygenase genes.Sequence analysis showed that these fragments were highly homologous to the known dioxygenase genes from pyrene-degrading Mycobacteria sp.

关 键 词：芘 微生物降解 16S rDNA鉴定 双加氧酶基因 分枝杆菌 

分 类 号：X172[环境科学与工程—环境科学]