Closed-loop identification of enzyme kinetics applying model-based design of experiments†

IF 3.4 3区 化学 Q2 CHEMISTRY, MULTIDISCIPLINARY
Leon Hennecke, Lucas Schaare, Mirko Skiborowski and Andreas Liese
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引用次数: 0

Abstract

Accurate kinetic models for enzyme catalysed reactions are integral to process development and optimisation. However, the collection of useful kinetic data is heavily dependent on the experimental design and execution. In order to reduce the limitations associated with traditional statistical design and manual experiments, this study introduces an integrated, automated approach to identifying kinetic models based on model-based optimal experimental design. The immobilised formate dehydrogenase of Candida boidinii catalyses the enzymatic reduction of NAD+ to NADH and is used as a model system. Continuous collection of UV/Vis data under steady-state conditions is employed to determine the kinetic parameters in a packed bed reactor. Automation of the experimental work was utilised in Python to compensate for the need for more time-consuming data collection. A completely automated closed-loop system was created and appropriate kinetic models for anticipating process dynamics were identified. The automated platform was able to identify the correct kinetic model out of eight candidate models with only 15 experiments. Further extension of the design space improved model discrimination and led to a properly parameterized kinetic model with sufficeintly high parameter precision for the conditions under examination.

Abstract Image

Abstract Image

应用基于模型的实验设计对酶动力学进行闭环识别
酶催化反应的精确动力学模型是工艺开发和优化不可或缺的一部分。然而,有用动力学数据的收集在很大程度上取决于实验设计和执行。为了减少传统统计设计和人工实验的局限性,本研究介绍了一种基于模型优化实验设计的综合自动化方法来确定动力学模型。固定化的 Candida boidinii 甲酸脱氢酶催化 NAD+ 酶促还原为 NADH,被用作模型系统。在稳态条件下连续收集紫外/可见光数据,以确定填料床反应器中的动力学参数。利用 Python 实现了实验工作的自动化,以弥补耗时较长的数据收集工作。创建了一个完全自动化的闭环系统,并确定了用于预测过程动态的适当动力学模型。自动化平台仅用 15 次实验就从 8 个候选模型中识别出了正确的动力学模型。设计空间的进一步扩展提高了模型的辨别能力,并产生了一个参数适当的动力学模型,其参数精度足够高,可用于所研究的条件。
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来源期刊
Reaction Chemistry & Engineering
Reaction Chemistry & Engineering Chemistry-Chemistry (miscellaneous)
CiteScore
6.60
自引率
7.70%
发文量
227
期刊介绍: Reaction Chemistry & Engineering is a new journal reporting cutting edge research into all aspects of making molecules for the benefit of fundamental research, applied processes and wider society. From fundamental, molecular-level chemistry to large scale chemical production, Reaction Chemistry & Engineering brings together communities of chemists and chemical engineers working to ensure the crucial role of reaction chemistry in today’s world.
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