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Induction characteristics of reductive dehalogenation in the ortho-halophenol-respiring bacterium, Anaeromyxobacter dehalogenans

机译:邻卤代苯酚呼吸细菌脱卤厌氧杆菌中还原性脱卤的诱导特性

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摘要

Anaeromyxobacter dehalogenans strain 2CP-C dehalogenates ortho-substituted di- and mono-halogenated phenols and couples this activity to growth. Reductive dehalogenation activity has been reported to be inducible, however, this process has not been studied extensively. In this study, the induction of reductive dehalogenation activity by strain 2CP-C is characterized. Constitutive 2-chlorophenol dechlorination activity occurs in non-induced fumarate-grown cells, with rates averaging 0.138 mumol of Cl- h(-1) mg of protein(-1). Once induced, these cultures dechlorinate 2-chlorophenol (2-CP) at rates as high as 116 mumol of Cl-1 h(-1) mg of protein(-1). Dechlorination of 2-CP is induced by phenol, 2-chlorophenol, 2, 4-dichlorophenol, 2, 5-dichlorophenol, 2, 6-dichlorophenol, and 2-bromophenol. Of the substrates tested, 2-bromophenol shows the highest induction potential, yielding double the 2-chlorophenol dechlorination rate when compared to other inducing substrates. No induced dechlorination is observed at concentrations less than 5 muM 2-CP. When fumarate cultures were diluted 100-fold, fumarate reduction rates were reduced roughly according to the dilution factor, while dechlorination rates were similar in fumarate grown cells amended with 2-CP and cells diluted 100-fold prior to the addition of chlorophenol. This indicates that the majority of the fumarate-grown cells in late log phase were not induced when exposed to inducing substrates such as 2-CP. This observation may have ramifications on the success of bioaugmentation using halorespiring bacteria, which traditionally relies on growing cultures using more readily utilized substrates. The rapid dechlorination rate and unique induction pattern also make strain 2CP-C a promising model organism for understanding the regulation of reductive dehalogenation at the enzymatic level.
机译:脱氧厌氧杆菌菌株2CP-C使邻位取代的二和一卤代苯酚脱卤,并使该活性与生长耦合。据报道还原性脱卤活性是可诱导的,但是,这一过程尚未得到广泛研究。在这项研究中,表征了菌株2CP-C诱导的还原脱卤活性。本构性的2-氯苯酚脱氯活性发生在未诱导的富马酸酯生长的细胞中,平均速率为0.138摩尔mol Cl-h(-1)mg蛋白(-1)。一旦诱导,这些培养物以高达116摩尔的Cl-1 h(-1)mg蛋白(-1)的速率对2-氯苯酚(2-CP)进行脱氯。苯酚,2-氯苯酚,2、4-二氯苯酚,2、5-二氯苯酚,2、6-二氯苯酚和2-溴苯酚可诱导2-CP脱氯。在测试的底物中,2-溴苯酚显示出最高的诱导潜力,与其他诱导性底物相比,产生的2-氯苯酚脱氯速率提高了一倍。浓度低于5μM2-CP时未观察到诱导的脱氯。当将富马酸酯培养物稀释100倍时,富马酸酯的还原率会根据稀释倍数大致降低,而富马酸酯生长的细胞经2-CP修饰后的脱氯速率与添加氯酚之前稀释100倍的细胞相似。这表明当暴露于诱导底物如2-CP时,对数后期富马酸酯生长的细胞没有被诱导。该观察结果可能对使用卤呼吸细菌进行生物增强的成功产生影响,该细菌传统上依赖于使用更易利用的底物生长的培养物。快速的脱氯速率和独特的诱导方式也使2CP-C菌株成为有前途的模型生物,可用于了解酶水平上还原性脱卤的调控。

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