title: High-level expression, purification and characterization of carbazole dioxygenase, a three components dioxygenase, of Pseudomonas GBS.5.
abstract: OBJECTIVE
  To investigate the conversion of carbazole into 2'-aminobiphenyl-2,3-diol using carbazole dioxygenase (CARDO) that is a multicomponent enzyme consisting of homotrimeric terminal oxygenases (CarAa), a ferredoxin (CarAc) and a ferredoxin reductase (CarAd) unit, encoded by the carAa, carAc and carAd genes, respectively.
  
  
  RESULTS
  The enzyme subunits containing a GST tag were expressed independently in E. coli. The expressed proteins were purified by one-step immobilized affinity chromatography and three purified proteins could reconstitute the CARDO activity in vitro and showed activity against carbazole as well as against wide range of polyaromatic compounds.
  
  
  CONCLUSIONS
  This method provides an efficient way to obtain an active carbazole dioxygenase with high yield, high purity and with activity against a wide range of polyaromatic compounds.
authors.affiliation: Department of Biotechnology, Jaypee Institute of Information Technology, A-10, Sector-62, Noida, 201301, U.P., India.
authors.forename: Samiya
authors.initials: S
authors.lastname: Khan
authors.affiliation: Biofuels Division & HOA Biotechnology Conversion Area, Indian Institute of Petroleum, Mohkampur, Dehradun, 248005, India.
authors.forename: D K
authors.initials: DK
authors.lastname: Adhikari
authors.affiliation: Department of Biotechnology, Jaypee Institute of Information Technology, A-10, Sector-62, Noida, 201301, U.P., India.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Gupta
authors.affiliation: Department of Biotechnology, Jaypee Institute of Information Technology, A-10, Sector-62, Noida, 201301, U.P., India. nidhi.gupta@jiit.ac.in.
authors.forename: Nidhi
authors.initials: N
authors.lastname: Gupta
mesh_terms: D002227:Carbazoles / Q000378:metabolism*; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D021122:Protein Subunits / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D002227:Carbazoles; D021122:Protein Subunits; D011994:Recombinant Proteins; C041514:carbazole; D049308:Dioxygenases
keywords: Biorefining; Carbazole; Dioxygenase; Fuel biorefining; GST-tag fusion; Polyaromatic compounds