Expression and functional analysis of three nicosulfuron-degrading enzymes from Bacillus subtilis YB1

To investigate the degradation activity of the manganese ABC transporter, vegetative catalase 1 and acetoin dehydrogenase E1 from Bacillus subtilis YB1, the proteins were prokaryotically expressed and purified. Assay results showed that the three enzymes were able to degrade nicosulfuron (2- (4,6-dimethoxypyrimidine-2-pyrimidinylcarbamoylaminosulfonyl) -N,N-dimethylnicotinamide), with vegetative catalase 1 exhibiting the highest activity. To further examine the degradation pathway, the degradation products of the three enzymes and the YB1 strain were detected by liquid chromatography-mass spectrometry(LC-MS). The nicosulfuron degradation products of the three enzymes were consistent with those of the YB1 strain, indicating the presence of two pathways: one due to cleavage of sulfonylurea bridges and ring-opening of 1-(4,6-dimethoxy-pyrimidin-2-yl)-3-(2-methyliminomethanesulfonyl-acetyl)-ureaas the pyrimidine ring, yielding the product; and the another due to cleavage of a sulfonylurea bridge, yielding 4,6-dihydroxy pyrimidine (111 m/z), 2-ylamine ?4,6-dimethoxy pyrimidine and ((4-(dimethycarbamoyl)pyridine-2-yl)sulfonyl)carbamic acid as products, which were further degraded to 4,6-dihydroxy pyrimidine and N,N-dimethyl-2-sulfamoyl-isonicotinamide. The above results reveal a major contribution of extracellular enzymes to the degradation of nicosulfuron by the YB1 strain. Our data help in elucidation of the mechanism of nicosulfuron bio-degradation and may facilitate the construction of engineered strains.