Expanding metabolic engineering algorithms using feasible space and shadow price constraint modules

• Added modules to impose multiple design criteria for engineering algorithms.
• Examples are provided to eliminate by-product secretion.
• Examples are provided to control coupling between product and biomass formation.
• Modules are tractable for genome-scale design.

While numerous computational methods have been developed that use genome-scale models to propose mutants for the purpose of metabolic engineering, they generally compare mutants based on a single criteria (e.g., production rate at a mutant׳s maximum growth rate). As such, these approaches remain limited in their ability to include multiple complex engineering constraints. To address this shortcoming, we have developed feasible space and shadow price constraint (FaceCon and ShadowCon) modules that can be added to existing mixed integer linear adaptive evolution metabolic engineering algorithms, such as OptKnock and OptORF. These modules allow strain designs to be identified amongst a set of multiple metabolic engineering algorithm solutions that are capable of high chemical production while also satisfying additional design criteria. We describe the various module implementations and their potential applications to the field of metabolic engineering. We then incorporated these modules into the OptORF metabolic engineering algorithm. Using an Escherichia coli genome-scale model (iJO1366), we generated different strain designs for the anaerobic production of ethanol from glucose, thus demonstrating the tractability and potential utility of these modules in metabolic engineering algorithms.