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IncP-1β Plasmid pGNB1 Isolated from a Bacterial Community from a Wastewater Treatment Plant Mediates Decolorization of Triphenylmethane Dyes▿

机译:从污水处理厂细菌群落中分离的IncP-1β质粒pGNB1介导三苯甲烷染料的脱色

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摘要

Plasmid pGNB1 was isolated from bacteria residing in the activated sludge compartment of a wastewater treatment plant by using a transformation-based approach. This 60-kb plasmid confers resistance to the triphenylmethane dye crystal violet and enables its host bacterium to decolorize crystal violet. Partial sequencing of pGNB1 revealed that its backbone is very similar to that of previously sequenced IncP-1β plasmids. The two accessory regions of the plasmid, one located downstream of the replication initiation gene trfA and the other located between the conjugative transfer modules Tra and Trb, were completely sequenced. Accessory region L1 contains a transposon related to Tn5501 and a gene encoding a Cupin 2 conserved barrel protein with an unknown function. The triphenylmethane reductase gene tmr and a truncated dihydrolipoamide dehydrogenase gene that is flanked by IS1071 and another putative insertion element were identified in accessory region L2. Subcloning of the pGNB1 tmr gene demonstrated that this gene is responsible for the observed crystal violet resistance phenotype and mediates decolorization of the triphenylmethane dyes crystal violet, malachite green, and basic fuchsin. Plasmid pGNB1 and the associated phenotype are transferable to the α-proteobacterium Sinorhizobium meliloti and the γ-proteobacterium Escherichia coli. This is the first report of a promiscuous IncP-1β plasmid isolated from the bacterial community from a wastewater treatment plant that harbors a triphenylmethane reductase gene. The pGNB1-encoded enzyme activity is discussed with respect to bioremediation of sewage polluted with triphenylmethane dyes.
机译:通过使用基于转化的方法,从位于污水处理厂活性污泥室中的细菌分离出质粒pGNB1。该60kb质粒赋予对三苯基甲烷染料结晶紫的抗性,并使它的宿主细菌使结晶紫脱色。 pGNB1的部分测序表明,其骨架与先前测序的IncP-1β质粒非常相似。质粒的两个辅助区域被完全测序,一个位于复制起始基因trfA的下游,另一个位于接合转移模块Tra和Trb之间。辅助区域L1含有与Tn5501有关的转座子和编码具有未知功能的Cupin 2保守桶蛋白的基因。在辅助区域L2中鉴定出三苯甲烷还原酶基因tmr和一个被IS1071和另一个推定的插入元件侧接的截短的二氢脂酰胺脱氢酶基因。 pGNB1 tmr基因的亚克隆表明,该基因负责观察到的结晶紫抗性表型,并介导了三苯基甲烷染料结晶紫,孔雀石绿和碱性品红的脱色。质粒pGNB1及其相关的表型可转移至α-变形杆菌Sinorhizobium meliloti和γ-变形杆菌Escherichia coli。这是从带有三苯甲烷还原酶基因的废水处理厂的细菌群落中分离的混杂IncP-1β质粒的首次报道。关于被三苯甲烷染料污染的污水的生物修复,讨论了pGNB1编码的酶活性。

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