Characterization of a novel oligomeric SGNH-arylesterase from Sinorhizobium meliloti 1021

Abstract

A novel oligomeric SGNH-arylesterase (Sm23) from Sinorhizobium meliloti 1021 was characterized using biochemical and biophysical methods. A sequence comparison of Sm23 with other SGNH members confirmed the presence of catalytic triad (Ser10, Asp187, and His190) and oxyanion holes (Ser10-Gly50-Asn90). The wild type enzyme was able to hydrolyze p-nitrophenyl acetate, α- and β-naphthyl acetate, while S10A mutant completely lost its activity. Structural properties of Sm23 were investigated using circular dichroism (CD), fluorescence, dynamic light scattering (DLS), chemical cross-linking, electron microscopy (EM), and time of flight (TOF) mass spectrometry. Furthermore, spherical or globular aggregates were observed with 1-butyl-3-methylimidazolium tetrafluoroborate, while amorphous aggregates were formed with 1-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide. © 2010 Elsevier B.V.