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Novel auto-inducing expression systems for the development of whole-cell biocatalysts

  • Applied Genetics and Molecular Biotechnology
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Abstract

Novel expression systems for the development of whole-cell biocatalysts were generated. Their novelty consists both in the host, Pseudomonas putida, and in the ability to auto-induce the expression of genes of interest at the exhaustion of the carbon source used for the biomass growth. The auto-induction relies on new expression vectors developed in this study and based on the activator TouR from Pseudomonas sp. OX1, which was shown to mediate the activation of target promoters in an effector-independent growth-phase-dependent manner when the carbon source is exhausted at the onset of the stationary phase. We validated the suitability of these expression systems through the production of (S)-styrene oxide by the styrene monooxygenase from Pseudomonas fluorescens ST. The yields of epoxides produced by these biocatalysts in flask experiments showed to be as efficient as those currently available based on inducible Escherichia coli systems. In addition, a larger scale of biomass production showed no reduction of biocatalysis efficiency. Therefore, the systems developed in this study constitute a valid alternative to current expression systems to use in bioconversion processes.

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Acknowledgments

This work was supported by Grants from MIUR-PRIN 2004 “Biocatalytic process development to produce bioproducts of interest”. We thank A. Baratto and V. Vigano’ for technical support and P. Barbieri for helpful discussion.

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Authors and Affiliations

  1. Department of Environmental Sciences, University of Milan-Bicocca, Milan, Italy

    Patrizia Di Gennaro, Silvia Ferrara & Giuseppina Bestetti

  2. Department of Organic and Industrial Chemistry, University of Milan, Milan, Italy

    Guido Sello

  3. DNA Editing Laboratory, Cancer Research UK, Clare Hall Laboratories, Blanche Lane South Mimms, Hertfordshire, EN6 3LD, UK

    Dafne Solera

  4. Department of Biomolecular Sciences and Biotechnology, University of Milan, Milan, Italy

    Enrica Galli, Francesco Renzi & Giovanni Bertoni

Authors
  1. Patrizia Di Gennaro
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  2. Silvia Ferrara
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  3. Giuseppina Bestetti
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  4. Guido Sello
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  5. Dafne Solera
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  6. Enrica Galli
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  7. Francesco Renzi
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  8. Giovanni Bertoni
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Correspondence to Patrizia Di Gennaro.

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Di Gennaro, P., Ferrara, S., Bestetti, G. et al. Novel auto-inducing expression systems for the development of whole-cell biocatalysts. Appl Microbiol Biotechnol 79, 617–625 (2008). https://doi.org/10.1007/s00253-008-1460-z

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  • DOI: https://doi.org/10.1007/s00253-008-1460-z

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