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Cloning of the Arthrobacter sp. FG1 dehalogenase genes and construction of hybrid pathways in Pseudomonas putida strains

  • Applied Genetics and Molecular Biotechnology
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Abstract

An Arthrobacter strain, able to utilize 4-chlorobenzoic acid as the sole carbon and energy source, was isolated and characterized. The first step of the catabolic pathway was found to proceed via a hydrolytic dehalogenation that leads to the formation of 4-hydroxybenzoic acid. The dehalogenase encoding genes (fcb) were sequenced and found highly homologous to and organized as those of other 4-chlorobenzoic acid degrading Arthrobacter strains. The fcb genes were cloned and successfully expressed in the heterologous host Pseudomonas putida PaW340 and P. putida KT2442 upper TOL, which acquired the ability to grow on 4-chlorobenzoic acid and 4-chlorotoluene, respectively. The cloned dehalogenase displayed a high specificity for para-substituted haloaromatics with affinity Cl > Br > I » F, in the order.

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Acknowledgments

This work was supported by the FIRB project n. RBAU015FL2 (MIUR, Rome) “Screening and characterization of ecofunctional genes and enzymes from high GC Gram-positive bacteria for ecological and bioremediation studies” of P.B. Part of this work was supported by a FEMS Short-term Fellowship given to F. Radice. F. Gorreta collaborated in the experimental work. We are also in debt with V. De Lorenzo, who kindly provided the P. putida KT2442 upper TOL strain, and with R. Eichenlaub, for providing pAS5.

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Authors and Affiliations

  1. Dipartimento di Biologia Strutturale e Funzionale, via Dunant 3, 21100, Varese, Italy

    Francesca Radice, Viviana Orlandi, Valentina Massa, Vanessa Battini & Paola Barbieri

  2. Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, via Celoria 26, 20133, Milan, Italy

    Giovanni Bertoni

  3. Chemische Mikrobiologie, Bergische Universität Wuppertal, Fachbereich C, Gaußstrasse 20, 42097, Wuppertal, Germany

    Walter Reineke

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  1. Francesca Radice
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  2. Viviana Orlandi
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  3. Valentina Massa
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  4. Vanessa Battini
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Correspondence to Paola Barbieri.

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Radice, F., Orlandi, V., Massa, V. et al. Cloning of the Arthrobacter sp. FG1 dehalogenase genes and construction of hybrid pathways in Pseudomonas putida strains. Appl Microbiol Biotechnol 75, 1111–1118 (2007). https://doi.org/10.1007/s00253-007-0906-z

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  • DOI: https://doi.org/10.1007/s00253-007-0906-z

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