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Cloning of a rhodococcal promoter using a transposon for dibenzothiophene biodesulfurization

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Abstract

The expression of biodesulfurization genes (dsz) in Rhodococcus erythropolis strain KA2-5-1 is repressed by sulfate which is the product of biodesulfurization. The application of a sulfate non-repressible promoter could be effective in enhancing biodesulfurization. A promoter-probe transposon was constructed using the promoterless, red-shifted green fluorescence protein gene (rsgfp). A 340 bp putative promoter element, designated kap1, was isolated from a strain KA2-5-1 recombinant that had shown high fluorescence intensity. The activity of kap1 was not affected by 1 mM sulfate. It gave about a 2-fold greater activity than the 16S ribosomal RNA promoter in R. erythropolis strain KA2-5-1 and is therefore useful for expressing desulfurization genes in rhodococcal strains.

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References

  • Goryshin IY, Jendrisak J, Hoffman LM, Meis R, Reznikoff WS (2000) Insertional transposon mutagenesis by electroporation of released Tn5 transposition complexes. Nat. Biotechnol. 18: 97-100.

    Google Scholar 

  • Gray KA, Pogrebinsky OS, Mrachko GT, Xi L, Montecello DJ, Squires CS (1996) Molecular mechanisms of biocatalytic desulfurization of fossil fuels. Nat. Biotechnol. 14: 1705-1709.

    Google Scholar 

  • Hirasawa K, Ishii Y, Kobayashi M, Koizumi K, Maruhashi K (2001) Improvement of desulfurization activity in Rhodococcus erythropolis KA2-5-1 by genetic engineering. Biosci. Biotechnol. Biochem. 65: 239-246.

    Google Scholar 

  • Izumi Y, Ohshiro T, Ogino H, Hine Y, Shimao M (1994) Selective desulfurization of dibenzothiophene by Rhodococcus erythropolis D-1. Appl. Environ. Microbiol. 60: 223-226.

    Google Scholar 

  • Ji Y-E, Colston MJ, Cox RA (1994) The ribosomal RNA (rrn) operons of fast-growing mycobacteria: primary and secondary structures and their relation to rrn operons of pathogenic slowgrowers. Microbiology 140: 2829-2840.

    Google Scholar 

  • Kobayashi M, Onaka T, Ishii Y, Konishi J, TakakiM, Okada H, Ohta Y, Koizumi K, Suzuki M (2000) Desulfurization of alkylated forms of both dibenzothiophene and benzothiophene by a single bacterial strain. FEMS Microbiol. Lett. 187: 123-126.

    Google Scholar 

  • Li MZ, Squires CH, Monticello DJ, Childs JD (1996) Genetic analysis of dsz promoter and associated regulatory regions of Rhodococcus erythropolis IGTS8. J. Bacteriol. 178: 6409-6418.

    Google Scholar 

  • Mangan MW, Meijer WG (2001) Random insertion mutagenesis of the intracellular pathogen Rhodococcus equi using transposomes. FEMS Microbiol. Lett. 205: 243-246.

    Google Scholar 

  • Matsui T, Noda K-i, Tanaka Y, Maruhashi K, Kurane R (2002) Recombinant Rhodococcus sp. strain T09 can desulfurize DBT in the presence of inorganic sulfate. Curr. Microbiol. 45: 240-244.

    Google Scholar 

  • Onaka T, Okumura K, Suzuki M (1997) Analysis of biodesulfurization products using solid-phase extraction. J. Chromatogr. Sci. 35: 417-422.

    Google Scholar 

  • Tang X, Lu BF, Pan Q (1999) A bifunctional transposon mini-Tn5gfp-km which can be used to select for promoter fusions and report gene expression levels in Agrobacterium tumefaciens. FEMS Microbiol. Lett. 179: 37-42.

    Google Scholar 

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Noda, Ki., Watanabe, K. & Maruhashi, K. Cloning of a rhodococcal promoter using a transposon for dibenzothiophene biodesulfurization. Biotechnology Letters 24, 1875–1882 (2002). https://doi.org/10.1023/A:1020926107636

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  • DOI: https://doi.org/10.1023/A:1020926107636

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