Goudar Chetan T, Harris Steve K, McInerney Michael J, Suflita Joseph M
Process and Technology Development, Bayer HealthCare, Biological Products Division, 800 Dwight Way, B56-A, Berkeley, CA 94710, USA.
J Microbiol Methods. 2004 Dec;59(3):317-26. doi: 10.1016/j.mimet.2004.06.013.
We present a simple method for estimating kinetic parameters from progress curve analysis of biologically catalyzed reactions that reduce to forms analogous to the Michaelis-Menten equation. Specifically, the Lambert W function is used to obtain explicit, closed-form solutions to differential rate expressions that describe the dynamics of substrate depletion. The explicit nature of the new solutions greatly simplifies nonlinear estimation of the kinetic parameters since numerical techniques such as the Runge-Kutta and Newton-Raphson methods used to solve the differential and integral forms of the kinetic equations, respectively, are replaced with a simple algebraic expression. The applicability of this approach for estimating Vmax and Km in the Michaelis-Menten equation was verified using a combination of simulated and experimental progress curve data. For simulated data, final estimates of Vmax and Km were close to the actual values of 1 microM/h and 1 microM, respectively, while the standard errors for these parameter estimates were proportional to the error level in the simulated data sets. The method was also applied to hydrogen depletion experiments by mixed cultures of bacteria in activated sludge resulting in Vmax and Km estimates of 6.531 microM/h and 2.136 microM, respectively. The algebraic nature of this solution, coupled with its relatively high accuracy, makes it an attractive candidate for kinetic parameter estimation from progress curve data.
我们提出了一种简单的方法,用于从生物催化反应的进程曲线分析中估计动力学参数,这些反应可简化为类似于米氏方程的形式。具体而言,兰伯特W函数用于获得描述底物消耗动态的微分速率表达式的显式、闭式解。新解的显式性质极大地简化了动力学参数的非线性估计,因为分别用于求解动力学方程的微分和积分形式的龙格-库塔法和牛顿-拉弗森法等数值技术被一个简单的代数表达式所取代。使用模拟和实验进程曲线数据相结合的方法,验证了该方法在估计米氏方程中Vmax和Km的适用性。对于模拟数据,Vmax和Km的最终估计值分别接近实际值1微摩尔/小时和1微摩尔,而这些参数估计的标准误差与模拟数据集的误差水平成正比。该方法还应用于活性污泥中细菌混合培养的氢气消耗实验,得到的Vmax和Km估计值分别为6.531微摩尔/小时和2.136微摩尔。该解的代数性质及其相对较高的准确性,使其成为从进程曲线数据估计动力学参数的一个有吸引力的候选方法。