Cloning and sequencing of the genes encoding 2-nitrotoluene dioxygenase from Pseudomonas sp. JS42

Gene
J V ParalesD T Gibson

Abstract

The first step in the metabolism of 2-nitrotoluene (2NT) by Pseudomonas sp. JS42 (JS42) is the addition of dioxygen to the aromatic nucleus of 2NT to form 3-methylcatechol with concomitant release of nitrite. This reaction is catalyzed by the three-component dioxygenase system 2-nitrotoluene 2,3-dioxygenase (2NTDO). We report here the cloning and nucleotide (nt) sequence of a 4912-basepair (bp) SacI DNA fragment from JS42 encoding all of the genes required for 2NTDO activity. Sequence analysis of the 4912-bp SacI DNA fragment revealed five open reading frames (ORFs). The amino acid (aa) sequences of the predicted polypeptides from these ORFs exhibit high homology to the aa sequences of polypeptides from other three-component dioxygenase systems. Based on aa sequence analyses, four of the peptides were designated Reductase2NT, Ferredoxin2NT, ISP alpha 2NT and ISP beta 2NT (ISP for iron-sulfur protein) with gene designations ntdAaAbAcAd. The predicted aa sequence from the remaining ORF (ORF2) had identity to ISP alpha subunits from other three-component dioxygenase systems but had a calculated molecular weight (M(r)) of 21,259, which is uncharacteristically small for ISP alpha subunits.

References

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Citations

Aug 19, 2004·Applied Microbiology and Biotechnology·L EyersSaid El Fantroussi
Dec 20, 2012·Applied Microbiology and Biotechnology·Xiao-Jun TianNing-Yi Zhou
Sep 22, 2005·Journal of Industrial Microbiology & Biotechnology·Kyung-Seon LeeRebecca E Parales
Mar 3, 2005·Biotechnology Letters·Hong LiuNing-Yi Zhou
Dec 6, 2012·Biodegradation·Deepak Singh, Gurunath Ramanathan
Jan 31, 2012·Applied and Environmental Microbiology·Christine A Rabinovitch-Deere, Rebecca E Parales
May 26, 2001·Applied and Environmental Microbiology·S D SicilianoC W Greer
Feb 2, 2002·Applied and Environmental Microbiology·Daniel J LessnerDavid T Gibson
Mar 7, 2006·Applied and Environmental Microbiology·Kou-San Ju, Rebecca E Parales
Apr 17, 2001·Bioscience, Biotechnology, and Biochemistry·J W NamT Omori
Jul 11, 2002·Journal of Bacteriology·Glenn R JohnsonJim C Spain
Oct 11, 2003·FEMS Microbiology Reviews·Joseph G LeahySuzanne M Morcomb
Feb 5, 2014·Biochemical and Biophysical Research Communications·Deepak SinghGurunath Ramanathan
Jan 18, 2008·FEMS Microbiology Ecology·Kristina UlrichDietrich Ewald
May 20, 2008·Advances in Applied Microbiology·R E ParalesJ L Ditty
May 29, 2010·Microbiology and Molecular Biology Reviews : MMBR·Kou-San Ju, Rebecca E Parales
Aug 26, 2020·BMC Bioinformatics·Ludovica LiguoriBruno Hay Mele

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