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Biotransformation of halophenols using crude cell extracts of Pseudomonas putida F6

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Abstract

Crude cell extracts of Pseudomonas putida F6 transformed 4-substituted fluoro-, chloro-, bromo- and iodo-phenol without the exogenous addition of cofactors. The rate of substrate consumption decreased with increasing substituent size (F>Cl>Br>I). Biotransformations resulted in greater than 95% utilisation of the halogenated substrate. Product accumulation was observed in incubations with 4-chloro, 4-bromo- and 4-iodo-phenol. These products were identified as the corresponding 4-substituted catechols. Transformation of 4-fluorophenol did not result in the accumulation of the corresponding catechol; however, manipulation of the reaction conditions by incorporation of ascorbic acid culminated in the formation of 4-fluorocatechol. Cell extracts of P. putida F6 also showed activity towards a 3-substituted phenol, namely 3-fluorophenol, resulting in the formation of a single product, 4-fluorocatechol.

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Acknowledgements

We wish to thank Dr. Zhi Li and Dongliang Chang (Institute of Biotechnology, ETH, Zurich, Switzerland) for carrying out LC-MS analysis, and Dr. Mike Casey and Kevin Conway (Department of Chemistry, University College Dublin, Dublin, Ireland) for carrying out GC-MS analysis. This work was supported by grants from the Enterprise Ireland Research Scholarship Programme (BR/1999/043), the Irish Environmental Protection Agency Contributory Scholarship Programme (2001-CS-(17/26) and the Wellcome Trust (055637/Z/98).

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Authors and Affiliations

  1. Department of Industrial Microbiology, University College Dublin, Belfield, Dublin 4, Republic of Ireland

    S. J. Brooks & E. M. Doyle

  2. Department of Biochemistry, Centre for Synthesis and Chemical Biology, Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Republic of Ireland

    C. Hewage & J. P. G. Malthouse

  3. Institute of Biotechnology, ETH Honggerberg, Zurich, Switzerland

    W. Duetz

  4. Department of Industrial Microbiology, Centre for Synthesis and Chemical Biology, Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Republic of Ireland

    K. E. O’ Connor

  5. Department of Industrial Microbiology, University College Dublin, Belfield, Dublin 4, Republic of Ireland

    K. E. O’ Connor

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  1. S. J. Brooks
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  2. E. M. Doyle
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  3. C. Hewage
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  4. J. P. G. Malthouse
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  5. W. Duetz
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  6. K. E. O’ Connor
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Correspondence to K. E. O’ Connor.

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Brooks, S.J., Doyle, E.M., Hewage, C. et al. Biotransformation of halophenols using crude cell extracts of Pseudomonas putida F6. Appl Microbiol Biotechnol 64, 486–492 (2004). https://doi.org/10.1007/s00253-003-1488-z

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  • DOI: https://doi.org/10.1007/s00253-003-1488-z

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