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恶臭假单胞菌TVA8对甲苯双加氧酶诱导的定量分析及三氯乙烯共代谢的动力学建模

Quantification of toluene dioxygenase induction and kinetic modeling of TCE cometabolism by Pseudomonas putida TVA8.

作者信息

Shingleton J T, Applegate B A, Baker A J, Sayler G S, Bienkowski P R

机构信息

Center for Environmental Biotechnology, The University of Tennessee, 676 Dabney Hall, Knoxville, Tennessee 37996-1605, USA.

出版信息

Biotechnol Bioeng. 2001 Dec;76(4):341-50. doi: 10.1002/bit.10083.

Abstract

As measured by the toluene-induced bioluminescent response of Pseudomonas putida TVA8 in batch experiments, toluene dioxygenase (Tod) enzyme activities are dependent on toluene concentration between 0 and 30 mg/L. To provide a measure of the Tod activity for use in Michaelis-Menten competitive-inhibition kinetics, a correlation between toluene concentration and induced Tod activity as measured by an induced bioluminescent response of P. putida TVA8 is presented as a nondimensional Tod activity parameter. A packed-bed, radial-flow bioreactor (RFB) using the bioreporter P. putida TVA8A serves as the model system for studying the effect of the enzyme activity parameter on model predictions of vapor-phase toluene oxidation and trichloroethylene (TCE) cometabolism. Mass balances were performed on a differential section of the RFB to describe the radial transport of vapor-phase toluene and TCE through a bulk gas phase and the concomitant biological reaction in a stationary biofilm phase. The finite-element Galerkin weak-statement formulation with first-order basis functions was used to find the optimum solution to the highly nonlinear, coupled equations. For this RFB system with toluene concentrations less than 1 mg/L in the bulk gas phase, the Tod activity parameter enables accurate predictions of steady-state TCE degradation rate (0.27 microg TCE/min).

摘要

在分批实验中,通过恶臭假单胞菌TVA8的甲苯诱导生物发光响应来测量,甲苯双加氧酶(Tod)的酶活性在0至30 mg/L的甲苯浓度范围内依赖于甲苯浓度。为了提供用于米氏竞争抑制动力学的Tod活性测量值,通过恶臭假单胞菌TVA8的诱导生物发光响应测量的甲苯浓度与诱导的Tod活性之间的相关性以无量纲Tod活性参数表示。使用生物报告菌恶臭假单胞菌TVA8A的填充床径向流生物反应器(RFB)作为模型系统,用于研究酶活性参数对气相甲苯氧化和三氯乙烯(TCE)共代谢模型预测的影响。对RFB的一个微分截面进行质量平衡,以描述气相甲苯和TCE通过主体气相的径向传输以及在固定生物膜相中的伴随生物反应。使用具有一阶基函数的有限元伽辽金弱陈述公式来找到高度非线性耦合方程的最优解。对于在主体气相中甲苯浓度小于1 mg/L的该RFB系统,Tod活性参数能够准确预测稳态TCE降解速率(0.27 μg TCE/分钟)。

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