Genomic structure and promoter analysis of the dsz operon for dibenzothiophene biodesulfurization from Gordonia alkanivorans RIPI90A
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The bacterium Gordonia alkanivorans RIPI90A has been previously reported as dibenzothiophene-desulfurizing strain. The present study provides a complete investigation of the dsz operon including dsz promoter analysis from desulfurization competent strain belonging to the genus Gordonia. PCR was used to amplify the dszABC genes and adaptor ligation-based PCR-walking strategy used to isolate the dsz promoter. Unlike the dsz operon of Rhodococcus erythropolis, the operon of RIPI90A was located on chromosome. Despite the remarkably high homology between dsz genes of G. alkanivorans RIPI90A and R. erythropolis IGST8, promoter sequences of the strains were not very similar. The dsz promoter of G. alkanivorans RIPI90A shows only 52.5% homology to that of R. erythropolis IGTS8 and Gordonia nitida. Deletion analysis of the dsz promoter from RIPI90A using luciferase as a reporter gene revealed that the dsz promoter was located in regions from −156 to −50.
KeywordsGordonia alkanivorans Biodesulfurization dsz operon PCR walking Promoter analysis Reporter gene
This work was supported by a fund from Iranian Research Institute of Petroleum Industry (R.I.P.I.). We gratefully thank M. Takeo for kindly donating the pRSG43 plasmid. The authors are grateful to Dr. S. Hoseinkhani for providing us with luciferase reporter gene and members of his laboratory for technical assistance in luciferase activity measurement.
- Denis-Larose C, Bergeron H, Labbe D, Greer CW, Hawari J, Grossman MJ, Sankey BM, Lau PCK (1998) Characterization of the basic replicon of Rhodococcus plasmid pSOX and development of a Rhodococcus–Escherichia coli shuttle vector. Appl Environ Microbiol 64:4363–4367Google Scholar
- Denome SA, Oldfield C, Nash LJ, Young KD (1994) Characterization of the desulfurization genes from Rhodococcus sp. strain IGTS8. J Bacteriol 176:6707–6716Google Scholar
- Emamzadeh A, Hosseinkhani S, Sadeghizadeh M, Nikkhah M, Chaichi MJ, Mortazavi M (2006) cDNA cloning, expression and homology modeling of luciferase from the firefly Lampyroidea maculata. J Biochem Mol Biolo 39:578–585Google Scholar
- Gallardo ME, Ferrandez A, Lorenzo VD, Garcia JL, Diaz E (1997) Designing recombinant Pseudomonas strains to enhance biodesulfurization. J Bacteriol 179:7156–7160Google Scholar
- Li ZM, Charles HS, Monticello DJ, Childs JD (1996) Genetic analysis of the dsz promoter and associated regulatory regions of Rhodococcus erythropolis IGTS8. J Bacteriol 178:6409–6418Google Scholar
- Park SJ, Lee I, Chang YK, Lee SY (2003) Desulfurization of dibenzothiophene and diesel oil by metabolically engineered E. coli. J Microbial Biotechnol 13:578–583Google Scholar
- Piddington CS, Kovacevich BR, Rambosek J (1995) Sequence and molecular characterization of a DNA region encoding the dibenzothiophene desulfurization operon of Rhodococcus sp. strain IGTS8. Appl Environ Microbiol 61:468–475Google Scholar
- Sambrook J, Russell DW (2001) Molecular cloning, a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
- Santos SCC, Alviano DS, Alviano CS, Pádula M, Leitao AC, Martins OB, Ribeiro CMS, Sassaki MYM, Matta CPS, Bevilaqua J, Sebastián GV, Seldin L (2006) Characterization of Gordonia sp. strain F.5.25.8 capable of dibenzothiophene desulfurization and carbazole utilization. Appl Microbiol Biotechnol 71:355–362CrossRefGoogle Scholar
- Vasicova P, Patek M, Nesvera J, Sahm H, Eikmanns B (1999) Analysis of the Corynebacterium glutamicum dapA promoter. J Bacteriol 181:6188–6191Google Scholar