title: Pseudomonas putida B2: a tod-lux bioluminescent reporter for toluene and trichloroethylene co-metabolism.
abstract: A tod-lux transcriptional fusion bioluminescent reporter strain, Pseudomonas putida B2, was developed to permit on-line analysis of trichloroethylene (TCE) transformation by toluene dioxygenase (todC1C2BA) in Pseudomonas putida F1. Strain B2 was exposed to toluene in growing and resting cell bioluminescence assays. The growing cells showed a direct correlation between bioluminescence and toluene concentration, while resting cells showed reproducible bioluminescence with repeated toluene exposures. In addition, P. putida B2 was encapsulated in alginate beads and used in a packed bed flow-through differential volume reactor. The TCE feed into the differential volume reactor was constant at 20 mg L-1 and toluene was pulsed in square-wave perturbations at 10 mg L-1. The system showed a direct correlation between the expression of the tod operon (as monitored by light output) and the co-metabolism of TCE. Approximately 20% of the TCE and 50% of the toluene was removed at a flow rate of 0.4 ml min-1. This approach allowed the on-line monitoring of tod gene expression and its relation to TCE biotransformation.
authors.affiliation: Center for Environmental Biotechnology, University of Tennessee, Knoxville 37932, USA.
authors.forename: B
authors.initials: B
authors.lastname: Applegate
authors.forename: C
authors.initials: C
authors.lastname: Kelly
authors.forename: L
authors.initials: L
authors.lastname: Lackey
authors.forename: J
authors.initials: J
authors.lastname: McPherson
authors.forename: S
authors.initials: S
authors.lastname: Kehrmeyer
authors.forename: F M
authors.initials: FM
authors.lastname: Menn
authors.forename: P
authors.initials: P
authors.lastname: Bienkowski
authors.forename: G
authors.initials: G
authors.lastname: Sayler
mesh_terms: D000464:Alginates / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics*; D001431:Bacteriological Techniques; D003001:Cloning, Molecular; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D008163:Luminescent Measurements; D016254:Mutagenesis, Insertional; D009876:Operon; D010105:Oxygenases / Q000235:genetics*; D010957:Plasmids; D016958:Pseudomonas putida / Q000235:genetics* / Q000378:metabolism*; D012097:Repressor Proteins; D014050:Toluene / Q000378:metabolism*; D015534:Trans-Activators; D014170:Transformation, Genetic; D014241:Trichloroethylene / Q000378:metabolism*
chemical_list: D000464:Alginates; D001426:Bacterial Proteins; D003470:Culture Media; D012097:Repressor Proteins; D015534:Trans-Activators; C064778:LuxR autoinducer binding proteins; D014241:Trichloroethylene; D014050:Toluene; D010105:Oxygenases; C023172:toluene dioxygenase