Control of substrate specificity by active-site residues in nitrobenzene dioxygenase

Applied and Environmental Microbiology
Kou-San Ju, R E Parales

Abstract

Nitrobenzene 1,2-dioxygenase from Comamonas sp. strain JS765 catalyzes the initial reaction in nitrobenzene degradation, forming catechol and nitrite. The enzyme also oxidizes the aromatic rings of mono- and dinitrotoluenes at the nitro-substituted carbon, but the basis for this specificity is not understood. In this study, site-directed mutagenesis was used to modify the active site of nitrobenzene dioxygenase, and the contribution of specific residues in controlling substrate specificity and enzyme performance was evaluated. The activities of six mutant enzymes indicated that the residues at positions 258, 293, and 350 in the alpha subunit are important for determining regiospecificity with nitroarene substrates and enantiospecificity with naphthalene. The results provide an explanation for the characteristic specificity with nitroarene substrates. Based on the structure of nitrobenzene dioxygenase, substitution of valine for the asparagine at position 258 should eliminate a hydrogen bond between the substrate nitro group and the amino group of asparagine. Up to 99% of the mononitrotoluene oxidation products formed by the N258V mutant were nitrobenzyl alcohols rather than catechols, supporting the importance of this hydrogen ...Continue Reading

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Citations

Jul 28, 2012·Biodegradation·Smriti RayuBrajesh K Singh
Feb 28, 2013·Protein Engineering, Design & Selection : PEDS·Janna ShainskyAyelet Fishman
May 29, 2010·Microbiology and Molecular Biology Reviews : MMBR·Kou-San Ju, Rebecca E Parales
Dec 9, 2008·Bioscience, Biotechnology, and Biochemistry·Hiromasa UchimuraHideaki Nojiri
Jan 28, 2014·Applied Microbiology and Biotechnology·Kalia Bernath-LevinAyelet Fishman
Sep 2, 2008·Current Opinion in Biotechnology·Shailendra SinghWilfred Chen
Feb 14, 2013·International Journal of Biological Macromolecules·Huseyin Bekir YildizHasim Yilmaz
Jan 1, 2012·Computational and Structural Biotechnology Journal·Adi Dror, Ayelet Fishman
Feb 5, 2014·Biochemical and Biophysical Research Communications·Deepak SinghGurunath Ramanathan
Jun 6, 2014·Physical Chemistry Chemical Physics : PCCP·Inacrist Geronimo, Piotr Paneth
Sep 8, 2011·Molecular Microbiology·Kou-San Ju, Rebecca E Parales
Jun 24, 2014·Journal of Chemical Theory and Computation·Anna PabisPiotr Paneth
Mar 15, 2014·The Journal of Physical Chemistry. B·Anna PabisPiotr Paneth

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