B. Applegate et al., PSEUDOMONAS-PUTIDA B2 - A TOD-LUX BIOLUMINESCENT REPORTER FOR TOLUENEAND TRICHLOROETHYLENE CO-METABOLISM, Journal of industrial microbiology & biotechnology, 18(1), 1997, pp. 4-9
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.