In silico experiments and applications of recombinant escherichia coli

Abstract

In this study, a novel in silico experiments for the systems biotechnological strategy towards the development of improved strains is proposed. Previous wet experimental studies have indicated that the enhanced production of commodity chemicals in metabolically engineered E. coli can be achieved by a variety of strategies based on genetic alterations and environmental changes. In the current study, such strategies are applied for production of succinic acids and are verified by resorting to a constraints-based flux analysis which provides more information on the internal flux distribution of the engineered E. coli. This process is initiated by constructing a genome-scale in silico model of recombinant E. coli, and then followed by quantifying metabolic fluxes in response to the flux variations for each amplified or deleted gene. The resultant flux distributions render it possible to investigate the effect of genetic perturbation, i.e., gene deletion, attenuation and enhancement, on internal flux patterns of E. coli. Finally, target genes for commodity chemicals were identified by analyzing the internal flux patterns. Possibly, the production of succinic acid can be more enhanced by appropriately manipulating such essential fluxes identified. Thus, the in silico experiments can provide a new insight into the metabolic engineering strategy for achieving higher succinic acid productivity.