Acta scientiarum naturalium Universitatis Pekinensis
LIU Dongqi1ZHU Shunni,NI Jinren
The gene encoding gentisate 1,2-dioxygenase from a soil-borne Gram-negative bacterium, Ralstonia solanacearum GMI1000, was cloned and overexpressed in E. coli. The resulting product incorporated a (His) 6 tag was purified to homogeneity from the harvested cell extracts by affinity chromatography. SDS-PAGE showed that the polypeptide exhibited an approximate molecular mass of 38 kDa. The optimal temperature and pH for gentisate cleavage catalyzed by the enzyme were 30 ℃ and 7.5, respectively. The Km of the enzyme was determined to be 56 μmol/L. The pI was 4.6-4.8. The active site of the gentisate 1,2-dioxygenase with gentisate was also modeled.
LIU Dongqi,ZHU Shunni,NI Jinren. Characterization of a Gentisate 1,2-dioxygenase from Ralstonia solanacearum GMI1000[J].Acta scientiarum naturalium Universitatis Pekinensis, 2007, 43(6): 828-833.
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