Purification and cloning of nicosulfuron-degrading enzymes from Bacillus subtilis YB1
Kang, Z.H.; Ren, C.C.; Zhang, J.L.; Dong, J.G.; Li, X.; Wei, X.J.
Prikladnaia Biokhimiia i Mikrobiologiia 50(1): 39-43
2014
ISSN/ISBN: 0555-1099 PMID: 25272750 Document Number: 673512
The nicosulfuron-degrading enzymes from Bacillus subtilis strain YB1 were purified and their genes were cloned. The proteins of bacterial culture filtrate were precipitated with ammonium sulfate or acetone. The extracellular proteins concentrated by acetone were purified from DEAE-Sepharose Fast Flow chromatography. The four protein peaks eluted from DEAE-column were separated and purified by native PAGE. Three components (P1-1, P3-2, P4-3) had nicosulfuron-degrading activity, and component P4-3 degradated 57.5% of this compound. The molecular weights of the components were 33.5, 54.8 and 37.0 kDa, respectively. The amino acid sequences of nicosulfuron-degrading enzymes from B. subtilis YB1 were determined by MALDI-TOF-MS, indicating these enzymes as manganese ABC transporter, vegetative catalase 1 and acetoin dehydrogenase E1, respectively. Using PCR amplification, genes 918, 1428, 1026 bp in size were detected for the enzymes studied.