bComamonas plasmids play important roles in shaping the phenotypes of their hosts and the adaptation of these hosts to changing environments, and understanding the evolutionary strategy of these plasmids is thus of great concern. In this study, the sequence of the 119-kb 3,5-dibromo-4-hydroxybenzonitrile-catabolizing plasmid pBHB from Comamonas sp. strain 7D-2 was studied and compared with those of three other Comamonas haloaromatic catabolic plasmids. Incompatibility group determination based on a phylogenetic analysis of 24 backbone gene proteins, as well as TrfA, revealed that these four plasmids all belong to the IncP-1 subgroup. Comparison of the four plasmids revealed a conserved backbone region and diverse genetic-load regions. The four plasmids share a core genome consisting of 40 genes (>50% similarities) and contain 12 to 50 unique genes each, most of which are xenobiotic-catabolic genes. Two functional reductive dehalogenase gene clusters are specifically located on pBHB, showing distinctive evolution of pBHB for haloaromatics. The higher catabolic ability of the bhbA2B2 cluster than the bhbAB cluster may be due to the transcription levels and the character of the dehalogenase gene itself rather than that of its extracytoplasmic binding receptor gene. The plasmid pBHB is riddled with transposons and insertion sequence (IS) elements, and ISs play important roles in the evolution of pBHB. The analysis of the origin of the bhb genes on pBHB suggested that these accessory genes evolved independently. Our work provides insights into the evolutionary strategies of Comamonas plasmids, especially into the adaptation mechanism employed by pBHB for haloaromatics. Comamonas species are ubiquitous in the environment and have been isolated from soil, mud, water, activated sludge, clinical samples, and organic pollutant-and heavy metal-contaminated environments (1). Comamonas species play an important role in matter cycling in nature and have been noted for their capabilities to degrade various aromatics (1-5). Interestingly, many Comamonas strains have been found to harbor plasmids, most of which have been confirmed to be related to the catabolism of xenobiotic aromatics (4, 6-8). For example, the plasmid pI2, responsible for the degradation of aniline/3-chloroaniline, was isolated from Comamonas testosteroni I2 (4, 9, 10). Another transferable plasmid, pTB30, which is involved in the mineralization of 3-chloroaniline, was isolated from C. testosteroni TB30 (9, 11). In addition, the plasmid pCNB1, encoding the catabolic pathway to convert 4-chloronitrobenzene to 5-chloro-4-hydroxy-2-oxovalerate, was sequenced in Comamonas sp. strain CNB-1 (12, 13). Recently, the plasmid pBHB was characterized in Comamonas sp. strain 7D-2. This plasmid is involved in the mineralization of the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile), which includes the reductive dehalogenation of the bromoxynil nitrile hydrolyte 3,5-dibromo-4-hydroxybenzoate (DBHB) (14).Plasmids are among the most important elements in the evolutio...
Microbial dehalogenation plays key roles in the biodegradation and detoxification of halogenated aromatics. Although the hydrolytic dehalogenation of halogenated aliphatic hydrocarbons and carboxylic acids has been extensively studied, there are few reports on the hydrolytic dehalogenation of halogenated aromatics. In this study, the aerobic strain BSQ-1, isolated from soils contaminated with halogenated aromatics in Jiangsu, China, was able to completely degrade 0.12 mM o-tetrachlorophthalonitrile in 84 hours in the presence of 2 mM sodium acetate trihydrate and released one equivalent of chlorine ions. MS-MS and NMR analysis revealed that o-tetrachlorophthalonitrile was dehalogenated to 4-hydroxyl-o-trichlorophthalonitrile. The dehalogenation of o-tetrachlorophthalonitrile could occur under both aerobic and anaerobic conditions, showing that the observed dehalogenation was a hydrolytic process. The optimal temperature and pH for o-tetrachlorophthalonitrile dehalogenation by strain BSQ-1 were 30°C and 8, respectively. It was found that 0.2 mM Zn 2+ , Mg 2+ or Co 2+ enhanced dehalogenation activity, whereas 0.2 mM Fe 3+ , Ni 2+ , Cu 2+ , Ca 2+ , or Pb 2+ inhibited dehalogenation activity. Strain BSQ-1 was able to dehalogenate not only
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