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Plasmid-mediated degradation of 4-chloronitrobenzene by newly isolated Pseudomonas putida strain ZWL73

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Abstract

A strain of Pseudomonasputida ZWL73 was isolated from soil contaminated with chloronitrobenzenes and identified by 16S rDNA sequencing. This bacterium released chloride and ammonia into the medium when grown on 4-chloronitrobenzene (4CNB) as the sole source of carbon, nitrogen and energy. A plasmid designated pZWL73 of approximately 100 kb in this strain was found to be responsible for 4CNB degradation. This was based on the fact that the plasmid-cured strains showed 4CNB phenotype and the 4CNB+ phenotype could be conjugally transferred. The cell-free extracts of strain ZWL73 exhibited chloronitrobenzene nitroreductase and 2-amino-5-chlorophenol 1, 6-dioxygenase (2A5CPDO) activities, but neither activity was found from that of the plasmid-cured strain. We have also cloned a 4.9-kb EcoRI fragment exhibiting 2A5CPDO activity. Sequencing results revealed β-subunit (cnbCa) and α subunit (cnbCb) of a meta-cleavage dioxygenase, which were subsequently expressed in E. coli with 2A5CPDO activity. The phylogenetic analysis suggested that 2A5CPDO may form a new subgroup in class III meta-cleavage dioxygenase with its close homologs.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC30230010). The authors would like to thank Peter Williams for Pseudomonas putida PaW340 and PaW1.

Author information

Correspondence to Ning-Yi Zhou.

Additional information

# D.Z. and H.L. equally contributed to this work.

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Zhen, D., Liu, H., Wang, S. et al. Plasmid-mediated degradation of 4-chloronitrobenzene by newly isolated Pseudomonas putida strain ZWL73. Appl Microbiol Biotechnol 72, 797–803 (2006). https://doi.org/10.1007/s00253-006-0345-2

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Keywords

  • Nitrobenzene
  • Pseudomonas Putida
  • Catabolic Gene
  • Protocatechuate
  • Cosmid Library