Kinetics of growth, oxygen uptake, and substrate utilization of wild type and genetically engineeredBurkholderia sp

Abstract

The gene (vgb) encoding the hemoglobin (VHb) ofVitreoscilla sp. was cloned intoBurkholderia sp. and the effect of VHb on the growth characteristics of genetically engineeredBurkholderia (YV1) were compared with wild typeBurkholderia (R34) using continuous flow reactors (chemostat) at various dilution rates under aerobic conditions. Batch oxygen uptake rate showed that YV1 has much higher oxygen uptake rate than R34 (i.e. 0.63 mg O2/g biomass/min vs. 1.43 mg O2/g biomass/min for R34 and YV1 respectively at a dilution rate of 1.2 day−1). Monod parameters, maximum growth rate (μmax) and half saturation coefficient (Ks) were found to be 7.03 day−1 and 691 mg/L for R34 respectively, compared to 5.49 day−1 and 404 mg/L for YV1 respectively. At low dilution rates (<2.5 day−1), when the substrate is present in low concentrations, the growth yield was much higher in YV1 (0.52) than in R34 (0.37). Although substrate utilization rates were similar between R34 and YV1, the latter showed much higher oxygen uptake rate than did R34 at all dilution rates. When the stability of VHb was tested on agar plates containing 40 μg/L of kanamycin and 100 μg/L of ampicillin,vgb gene containing VHb plasmid in YV1 was stable over 82 days. When survivability under oxygen limited conditions was tested, R34 survived only for 11 days whereas YV1 survived over 25 days in liquid media; in agar plate experiments, R34 did not survive more than 40 days whereas more than 75% of YV1 survived over 110 days.