Folsom B R, Chapman P J, Pritchard P H
Technical Resources, Inc., Gulf Breeze, Florida 32561.
Appl Environ Microbiol. 1990 May;56(5):1279-85. doi: 10.1128/aem.56.5.1279-1285.1990.
Intact cells of Pseudomonas cepacia G4 completely degraded trichloroethylene (TCE) following growth with phenol. Degradation kinetics were determined for both phenol, used to induce requisite enzymes, and TCE, the target substrate. Apparent Ks and Vmax values for degradation of phenol by cells were 8.5 microM and 466 nmol/min per mg of protein, respectively. At phenol concentrations greater than 50 microM, phenol degradation was inhibited, yielding an apparent second-order inhibitory value, KSI, of 0.45 mM as modeled by the Haldane expression. A partition coefficient for TCE was determined to be 0.40 +/- 0.02, [TCEair]/[TCEwater], consistent with Henry's law. To eliminate experimental problems associated with TCE volatility and partitioning, a no-headspace bottle assay was developed, allowing for direct and accurate determinations of aqueous TCE concentration. By this assay procedure, apparent Ks and Vmax values determined for TCE degradation by intact cells were 3 microM and 8 nmol/min per mg of protein, respectively. Following a transient lag period, P. cepacia G4 degraded TCE at concentrations of at least 300 microM with no apparent retardation in rate. Consistent with Ks values determined for degradation, TCE significantly inhibited phenol degradation.
洋葱伯克霍尔德菌G4的完整细胞在以苯酚为底物生长后能完全降解三氯乙烯(TCE)。分别测定了用于诱导所需酶的苯酚以及目标底物TCE的降解动力学。细胞降解苯酚的表观Ks值和Vmax值分别为8.5微摩尔和每毫克蛋白质466纳摩尔/分钟。当苯酚浓度大于50微摩尔时,苯酚降解受到抑制,根据哈代表达式建模,其表观二级抑制值KSI为0.45毫摩尔。TCE的分配系数确定为0.40±0.02,即[TCE空气]/[TCE水],符合亨利定律。为消除与TCE挥发性和分配相关的实验问题,开发了一种无顶空瓶试验,可直接准确测定水相中TCE的浓度。通过该试验程序,完整细胞降解TCE的表观Ks值和Vmax值分别为3微摩尔和每毫克蛋白质8纳摩尔/分钟。经过短暂的滞后期后,洋葱伯克霍尔德菌G4能降解至少300微摩尔浓度的TCE,且速率无明显延迟。与降解测定的Ks值一致,TCE显著抑制苯酚降解。