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首页> 外文期刊>Applied Microbiology >Role of IncP-1β Plasmids pWDL7::rfp and pNB8c in Chloroaniline Catabolism as Determined by Genomic and Functional Analyses
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Role of IncP-1β Plasmids pWDL7::rfp and pNB8c in Chloroaniline Catabolism as Determined by Genomic and Functional Analyses

机译：基因组和功能分析确定IncP-1β质粒pWDL7 :: rfp和pNB8c在氯苯胺代谢中的作用

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Broad-host-range catabolic plasmids play an important role in bacterial degradation of man-made compounds. To gain insight into the role of these plasmids in chloroaniline degradation, we determined the first complete nucleotide sequences of an IncP-1 chloroaniline degradation plasmid, pWDL7:: rfp and its close relative pNB8c, as well as the expression pattern, function, and bioaugmentation potential of the putative 3-chloroaniline (3-CA) oxidation genes. Based on phylogenetic analysis of backbone proteins, both plasmids are members of a distinct clade within the IncP-1β subgroup. The plasmids are almost identical, but whereas pWDL7:: rfp carries a duplicate inverted catabolic transposon, Tn 6063 , containing a putative 3-CA oxidation gene cluster, dcaQTA1A2BR , pNB8c contains only a single copy of the transposon. No genes for an aromatic ring cleavage pathway were detected on either plasmid, suggesting that only the upper 3-CA degradation pathway was present. The dcaA1A2B gene products expressed from a high-copy-number vector were shown to convert 3-CA to 4-chlorocatechol in Escherichia coli . Slight differences in the dca promoter region between the plasmids and lack of induction of transcription of the pNB8c dca genes by 3-CA may explain previous findings that pNB8C does not confer 3-CA transformation. Bioaugmentation of activated sludge with pWDL7:: rfp accelerated removal of 3-CA, but only in the presence of an additional carbon source. Successful bioaugmentation requires complementation of the upper pathway genes with chlorocatechol cleavage genes in indigenous bacteria. The genome sequences of these plasmids thus help explain the molecular basis of their catabolic activities.
机译：广泛的宿主分解代谢质粒在人造化合物的细菌降解中起重要作用。为了深入了解这些质粒在氯苯胺降解中的作用，我们确定了IncP-1氯苯胺降解质粒pWDL7 :: rfp及其近亲pNB8c的第一个完整核苷酸序列，以及表达模式，功能和生物增强作用潜在的3-氯苯胺（3-CA）氧化基因的潜力。基于主链蛋白的系统发育分析，两个质粒都是IncP-1β子群内不同进化枝的成员。质粒几乎相同，但是pWDL7 :: rfp携带一个重复的反向分解代谢转座子Tn 6063，其中包含一个推定的3-CA氧化基因簇dcaQTA1A2BR，pNB8c仅包含一个转座子拷贝。在任一质粒上均未检测到芳香环裂解途径的基因，表明仅存在上部3-CA降解途径。从高拷贝数载体表达的dcaA1A2B基因产物显示出在大肠杆菌中将3-CA转化为4-氯儿茶酚。质粒之间的dca启动子区域略有差异，并且3-CA对pNB8c dca基因的转录缺乏诱导，这可以解释先前的发现，即pNB8C不赋予3-CA转化。用pWDL7 :: rfp对活性污泥进行生物强化可加速3-CA的去除，但仅在存在其他碳源的情况下。成功的生物增强需要在本地细菌中用氯邻苯二酚裂解基因对上游途径基因进行补充。因此，这些质粒的基因组序列有助于解释其分解代谢活性的分子基础。

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