Molecular cloning and partial characterization of the pathway for aniline degradation inPseudomonas sp. strain CIT1

Meyers, Nicholas

doi:10.1007/bf01571099

Cited by 12 publications

(11 citation statements)

References 20 publications

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“…All the tdn genes are essential for the conversion of aniline to catechol. A number of catabolic plasmids, such as pCIT1, pTDN1, and pYA1, that can degrade aniline have been described previously (2,17,38,48).In contrast to aniline, which is rapidly metabolized, chloroaniline is more persistent in the environment (27, 55). Therefore, many efforts have been undertaken to isolate bacteria capable of degrading chlorinated anilines.…”

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Genetic Diversity among 3-Chloroaniline- and Aniline-Degrading Strains of the Comamonadaceae

Boon

Goris

Vos

et al. 2001

Appl Environ Microbiol

154

141

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We examined the diversity of the plasmids and of the gene tdnQ, involved in the oxidative deamination of aniline, in five bacterial strains that are able to metabolize both aniline and 3-chloroaniline (3-CA). Three strains have been described and identified previously, i.e., Comamonas testosteroni I2 and Delftia acidovorans CA28 and BN3.1. Strains LME1 and B8c were isolated in this study from linuron-treated soil and from a wastewater treatment plant, respectively, and were both identified as D. acidovorans. Both Delftia and Comamonas belong to the family Comamonadaceae. All five strains possess a large plasmid of ca. 100 kb, but the plasmids from only four strains could be transferred to a recipient strain by selection on aniline or 3-CA as a sole source of carbon and/or nitrogen. Plasmid transfer experiments and Southern hybridization revealed that the plasmid of strain I2 was responsible for total aniline but not 3-CA degradation, while the plasmids of strains LME1 and B8c were responsible only for the oxidative deamination of aniline. Several transconjugant clones that had received the plasmid from strain CA28 showed different degradative capacities: all transconjugants could use aniline as a nitrogen source, while only some of the transconjugants could deaminate 3-CA. For all four plasmids, the IS1071 insertion sequence of Tn5271 was found to be located on a 1.4-kb restriction fragment, which also hybridized with the tdnQ probe. This result suggests the involvement of this insertion sequence element in the dissemination of aniline degradation genes in the environment. By use of specific primers for the tdnQ gene from Pseudomonas putida UCC22, the diversity of the PCR-amplified fragments in the five strains was examined by denaturing gradient gel electrophoresis (DGGE). With DGGE, three different clusters of the tdnQ fragment could be distinguished. Sequencing data showed that the tdnQ sequences of I2, LME1, B8c, and CA28 were very closely related, while the tdnQ sequences of BN3.1 and P. putida UCC22 were only about 83% identical to the other sequences. Northern hybridization revealed that the tdnQ gene is transcribed only in the presence of aniline and not when only 3-CA is present.For many years, anilines and chloroanilines have been among the most important industrially produced amines. They are used widely in the production of polyurethanes, rubber, azo dyes, drugs, photographic chemicals, varnishes, and pesticides (20,29). As a consequence of this widespread use, they are detected in wastewaters (31, 49). Moreover, chloroanilines have been found in waters as a consequence of the transformation of frequently used acetamide and urea herbicides (31). These toxic and recalcitrant compounds are considered important environmental pollutants (37) and are subject to legislative control by the 76/464/EEC Directive (13) and by the Priority Pollutant List of the U.S. Environmental Protection Agency (15).In aquatic environments, the major way to remove aniline is through biodegradation (20,35). The first step of t...

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Genetic Diversity among 3-Chloroaniline- and Aniline-Degrading Strains of the Comamonadaceae

Boon

Goris

Vos

et al. 2001

Appl Environ Microbiol

154

141

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“…Simple anilines such as aniline and monosubstituted anilines are known to disappear from the environment mainly via biodegradation (8,9). To understand the mechanisms of aniline biodegradation, many aniline-degrading bacteria have been isolated and characterized (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25).…”

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Function of a Glutamine Synthetase-Like Protein in Bacterial Aniline Oxidation via γ-Glutamylanilide

et al. 2013

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Acinetobacter sp. strain YAA has five genes (atdA1 to atdA5) involved in aniline oxidation as a part of the aniline degradation gene cluster. From sequence analysis, the five genes were expected to encode a glutamine synthetase (GS)-like protein (AtdA1), a glutamine amidotransferase-like protein (AtdA2), and an aromatic compound dioxygenase (AtdA3, AtdA4, and AtdA5) (M. Takeo, T. Fujii, and Y. Maeda, J. Ferment. Bioeng. 85:17-24, 1998). A recombinant Pseudomonas strain harboring these five genes quantitatively converted aniline into catechol, demonstrating that catechol is the major oxidation product from aniline. To elucidate the function of the GS-like protein AtdA1 in aniline oxidation, we purified it from recombinant Escherichia coli harboring atdA1. The purified AtdA1 protein produced gamma-glutamylanilide (␥-GA) quantitatively from aniline and L-glutamate in the presence of ATP and MgCl 2 . This reaction was identical to glutamine synthesis by GS, except for the use of aniline instead of ammonia as the substrate. Recombinant Pseudomonas strains harboring the dioxygenase genes (atdA3 to atdA5) were unable to degrade aniline but converted ␥-GA into catechol, indicating that ␥-GA is an intermediate to catechol and a direct substrate for the dioxygenase. Unexpectedly, a recombinant Pseudomonas strain harboring only atdA2 hydrolyzed ␥-GA into aniline, reversing the ␥-GA formation by AtdA1. Deletion of atdA2 from atdA1 to atdA5 caused ␥-GA accumulation from aniline in recombinant Pseudomonas cells and inhibited the growth of a recombinant Acinetobacter strain on aniline, suggesting that AtdA2 prevents ␥-GA accumulation that is harmful to the host cell.

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“…A few of these plasmids have recently been completely sequenced: pJP4, pEST4011 [2,4-dichlorophenoxyacetic acid degradation (58,61)], pUO1 [haloacetate degradation (44)], pADP-1 [atrazine degradation (33)], pA81 [chlorobenzoate degradation (23)], and pCNB1 [4-chloronitrobenzene degradation (32)]; details regarding most of these plasmid sequences are also summarized in a recent review (60). While several catabolic plasmids that encode (partial) (chloro)aniline degradation pathways have been described, such as pCIT1 (2,34), pTDN1 (18,40), pYA1 (17), pNB1, pNB2, pNB8c, pC1 (7), and pWDL7 and pTB30 (14), their complete genome sequences have not yet been reported. Some of these plasmids were shown to transfer the ability to use 3-chloroaniline (3-CA) as the sole nitrogen or occasionally the sole carbon source to other hosts (3,6,7,14).…”

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Role of IncP-1β Plasmids pWDL7:: rfp and pNB8c in Chloroaniline Catabolism as Determined by Genomic and Functional Analyses

Król

Penrod

McCaslin

et al. 2012

Appl Environ Microbiol

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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-1␤ subgroup. The plasmids are almost identical, but whereas pWDL7::rfp carries a duplicate inverted catabolic transposon, Tn6063, 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.

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Molecular cloning and partial characterization of the pathway for aniline degradation inPseudomonas sp. strain CIT1

Cited by 12 publications

References 20 publications

Genetic Diversity among 3-Chloroaniline- and Aniline-Degrading Strains of the Comamonadaceae

Genetic Diversity among 3-Chloroaniline- and Aniline-Degrading Strains of the Comamonadaceae

Function of a Glutamine Synthetase-Like Protein in Bacterial Aniline Oxidation via γ-Glutamylanilide

Role of IncP-1β Plasmids pWDL7:: rfp and pNB8c in Chloroaniline Catabolism as Determined by Genomic and Functional Analyses

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