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Genetic and Functional Characterization of a Salicylate 1-monooxygenase Located on an Integrative and Conjugative Element (ICE) in Pseudomonas stutzeri AJR13

  • Microbial Genetics, Genomics and Molecular Biology
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Abstract

Pseudomonas stutzeri strain AJR13 was isolated for growth on the related compounds biphenyl (BPH) and diphenylmethane (DPM). The BPH and DPM degradative pathway genes are present on an integrative and conjugative element (ICE) in the chromosome. Examination of the genome sequence of AJR13 revealed a gene encoding a salicylate 1-monooxygenase (salA) associated with the ICE even though AJR13 did not grow on salicylate. Transfer of the ICE to the well-studied Pseudomonas putida KT2440 resulted in a KT2440 strain that could grow on salicylate. Knockout mutagenesis of the salA gene on the ICE in KT2440 eliminated the ability to grow on salicylate. Complementation of the knockout with the cloned salA gene restored growth on salicylate. Transfer of the cloned salA gene under control of the lac promoter to KT2440 resulted in a strain that could grow on salicylate. Heterologous expression of the salA gene in E. coli BL21 DE3 resulted in the production of catechol from salicylate, confirming that it is indeed a salicylate 1-monooxygenase. Interestingly, transfer of the cloned salA gene under control of the lac promoter to AJR13 resulted in a strain that could now grow on salicylate, suggesting that gene expression for the downstream catechol pathway is intact.

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Acknowledgements

Igor Ivanovski acknowledges support from NIEHS grant P42-ES004911 under the Superfund Research Program.

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Authors and Affiliations

  1. Department of Biology, St. Joseph’s University, Patchogue, NY, 11772, USA

    Igor Ivanovski

  2. Department of Biochemistry and Microbiology, School of Environmental and Biological Sciences, Rutgers University, New Brunswick, NJ, 08901, USA

    Igor Ivanovski & Gerben J. Zylstra

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  1. Igor Ivanovski
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  2. Gerben J. Zylstra
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II conceptualization (supporting); writing—original draft (lead); formal analysis (equal); writing—review and editing (lead). GJZ conceptualization (lead); writing—original draft (supporting); formal analysis (equal); writing—review and editing (supporting).

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Correspondence to Igor Ivanovski.

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Ivanovski, I., Zylstra, G.J. Genetic and Functional Characterization of a Salicylate 1-monooxygenase Located on an Integrative and Conjugative Element (ICE) in Pseudomonas stutzeri AJR13. J Microbiol. 61, 1025–1032 (2023). https://doi.org/10.1007/s12275-023-00093-x

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  • DOI: https://doi.org/10.1007/s12275-023-00093-x

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