PSEUDOMONAS-PUTIDA B2 - A TOD-LUX BIOLUMINESCENT REPORTER FOR TOLUENEAND TRICHLOROETHYLENE CO-METABOLISM

A tod-lux transcriptional fusion bioluminescent reporter strain, Pseud omonas putida B2, was developed to permit on-line analysis of trichlor oethylene (TCE) transformation by toluene dioxygenase (todC1C2BA) in P seudomonas 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 toluen e exposures. In addition, P, putida B2 was encapsulated in alginate be ads 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-l, 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 0.4mlmin(-1). This approach allowed the on-line moni toring of fod gene expression and its relation to TCE biotransformatio n.