Expression, purification and substrate specificities of 3-nitrotoluene dioxygenase from Diaphorobacter sp. strain DS2.
3-Nitotoluene dioxygenase (3-NTDO) is the first enzyme in the degradation pathway of 3-nitrotoluene (3-NT) by Diaphorobacter sp. strain DS2. The complete gene sequences of 3-NTDO were PCR amplified from genomic DNA of Diaphorobacter sp., cloned, sequenced and expressed. The 3-NTDO gene revealed a multi component structure having a reductase, a ferredoxin and two oxygenase subunits. Clones expressing the different subunits were constructed in pET21a expression vector system and overexpressed in E. coli BL21(DE3) host. Each subunit was individually purified separately to homogeneity. The active recombinant enzyme was reconstituted in vitro by mixing all three purified subunits. The reconstituted recombinant enzyme could catalyse biotransformations on a variety of organic aromatics.
['Singh|Deepak|D|', 'Kumari|Archana|A|', 'Ramaswamy|S|S|', 'Ramanathan|Gurunath|G|']
D001426:Bacterial Proteins D042621:Comamonadaceae D004136:Dinitrobenzenes D049308:Dioxygenases D004591:Electrophoresis, Polyacrylamide Gel D004926:Escherichia coli D005288:Ferredoxins D015964:Gene Expression Regulation, Bacterial D015971:Gene Expression Regulation, Enzymologic D006841:Hydrocarbons, Aromatic D015394:Molecular Structure D010088:Oxidoreductases D010105:Oxygenases D021122:Protein Subunits D011994:Recombinant Proteins D013379:Substrate Specificity D014050:Toluene
Diaphorobacter 3-Nitotoluene dioxygenase (3-NTDO) Biodegradation Nitrotoluene (NT) Rieske dioxygenase