原废水培养基分离活性污泥中的苯酚降解细菌

用未经处理的含酚焦化工业废水为基础配制培养基，采用平板涂布法，在10^-5稀释度下得到100个单菌落分离物．它们均能在以苯酚(500-800mg L^-1)为唯一碳源的无机盐培养基上生长，其16S rDNA基因的ARDBA多态性分析将这些分离物划分成4个分类操作单元(OTUs)．各单元代表株的16S rRNA基因序列分析和检索结果表明，97个分离物与Alcaligenes faecalis同源性达99％，2个分离物分别与Arthrobacter nicotianae和Klebsiella sp.99％同源．另一个分离物与Ochrobactrum sp．96％同源．苯酚经化酶大亚基基因(LmPHs)PCR扩增表明，除类似Arthrobacter nicotianae的菌株外，其它3个分类操作单元的降酚菌都能利用多组分苯酚经化酶代谢途径进行酚代谢．图2表2参11

ISOLATION OF NOVEL PHENOL-DEGRADING BACTERIA FROM ACTIVATED SLUDGE USING FEED WATER MEDIUM (FWM)

100 isolates were obtained from feed water solid media using spread-plate method with diluted activated sludge ( 10 -5 dilution rate). All the isolates were capable of growing on a synthetic medium with phenol (500 ~ 800 mg L -1 ) as a sole carbon source and classified into 4 operational taxoriomic units (OTUs) by amplified ribosomal DNA restriction analysis (ARDRA). The full-length 16S rDNA gene of one representative strain for each OTU was amplified, cloned, sequenced and blasted against the GenBank database. The results showed that 97 isolates were closely related to Alcaligenes faecalis with 99% homology; two isolates showed 99% identity to the sequence of Arthrobacter nicotianae and Klebsiella sp. respectively; and the last one exhibited 96% homology with Ochrobactrum sp. The specific amplification of the gene encoding the large subunit of the multi-component phenol hydroxylase (LmPHs) revealed that three out of four OTUs employed the metabolic pathway catalyzed by multicomponent phenol hydroxylase to degrade phenol, while the specific amplification from the Arthrobacter nicotianae-like strain did not show any products, indicating a possibly different phenol-degrading pathway. Fig 2, Tab 2, Ref 11