Robust systematic approaches for the metabolic engineering of cell factories remain elusive. The available models for predicting phenotypical responses and mechanisms are incomplete, particularly within the context of compound toxicity that can be a significant impediment to achieving high yields of a target product. This study describes a Multi-Omic Based Production Strain Improvement (MOBpsi) strategy that is distinguished by integrated time-resolved systems analyses of fed-batch fermentations. As a case study, MOBpsi was applied to improve the performance of an Escherichia coli cell factory producing the commodity chemical styrene. Styrene can be bio-manufactured from phenylalanine via an engineered pathway comprised of the enzymes phenylalanine ammonia lyase and ferulic acid decarboxylase. The toxicity, hydrophobicity, and volatility of styrene combine to make bio-production challenging. Previous attempts to create styrene tolerant E. coli strains by targeted genetic interventions have met with modest success. Application of MOBpsi identified new potential targets for improving performance, resulting in two host strains (E. coli NST74ΔaaeA and NST74ΔaaeA cpxP_o) with increased styrene production. The best performing re-engineered chassis, NST74ΔaaeA cpxP_o, produced ∼3 × more styrene and exhibited increased viability in fed-batch fermentations. Thus, this case study demonstrates the utility of MOBpsi as a systematic tool for improving the bio-manufacturing of toxic chemicals.

用于细胞工厂代谢工程的稳健系统方法仍然难以捉摸。用于预测表型反应和机制的可用模型是不完整的，特别是在化合物毒性的背景下，这可能是实现目标产品高产量的重大障碍。本研究描述了一种基于多组学的生产菌株改进 (MOB psi ) 策略，该策略通过补料分批发酵的集成时间分辨系统分析来区分。作为一个案例研究，MOB psi被应用于提高生产商品化学苯乙烯的大肠杆菌细胞工厂的性能。苯乙烯可以通过苯丙氨酸生物制造由苯丙氨酸解氨酶和阿魏酸脱羧酶组成的工程途径。苯乙烯的毒性、疏水性和挥发性相结合，使生物生产具有挑战性。以前通过有针对性的基因干预来产生耐苯乙烯的大肠杆菌菌株的尝试取得了一定的成功。MOB psi的应用确定了提高性能的新潜在目标，导致两种宿主菌株（大肠杆菌NST74Δ aaeA和 NST74Δ aaeA cpxP _o）的苯乙烯产量增加。性能最佳的重新设计机箱，NST74Δ aaeA cpxP _o，产生了约 3 倍的苯乙烯，并在补料分批发酵中表现出更高的活力。因此，本案例研究证明了 MOB psi作为改进有毒化学品生物制造的系统工具的效用。