Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA.
Metab Eng. 2010 Mar;12(2):150-60. doi: 10.1016/j.ymben.2009.07.006. Epub 2009 Jul 29.
We have developed a reactor-scale model of Escherichia coli metabolism and growth in a 1000 L process for the production of a recombinant therapeutic protein. The model consists of two distinct parts: (1) a dynamic, process specific portion that describes the time evolution of 37 process variables of relevance and (2) a flux balance based, 123-reaction metabolic model of E. coli metabolism. This model combines several previously reported modeling approaches including a growth rate-dependent biomass composition, maximum growth rate objective function, and dynamic flux balancing. In addition, we introduce concentration-dependent boundary conditions of transport fluxes, dynamic maintenance demands, and a state-dependent cellular objective. This formulation was able to describe specific runs with high-fidelity over process conditions including rich media, simultaneous acetate and glucose consumption, glucose minimal media, and phosphate depleted media. Furthermore, the model accurately describes the effect of process perturbations--such as glucose overbatching and insufficient aeration--on growth, metabolism, and titer.
我们开发了一个大肠杆菌代谢和生长的反应器规模模型，该模型用于在 1000L 工艺中生产重组治疗蛋白。该模型由两个不同的部分组成：（1）一个动态的、特定于过程的部分，描述了 37 个相关过程变量的时间演变；（2）一个基于通量平衡的大肠杆菌代谢的 123 反应代谢模型。该模型结合了几种以前报道的建模方法，包括生长速率依赖的生物质组成、最大生长速率目标函数和动态通量平衡。此外，我们引入了浓度依赖的运输通量边界条件、动态维持需求和状态依赖的细胞目标。这种表述能够在包括丰富培养基、同时消耗乙酸盐和葡萄糖、葡萄糖最小培养基和磷酸盐耗尽培养基在内的过程条件下，对特定运行进行高精度描述。此外，该模型还准确地描述了过程扰动（如葡萄糖过量进料和通风不足）对生长、代谢和滴度的影响。
