(441e) Expansions to Dynamic Substrate Analysis Builder (DSAB) and Tests Utilizing Mycoplasma Genitalium, Escherichia Coli and Mus Musculus

Our past work has focused on the creation of the Dynamic Substrate Analysis Builder (DSAB), a tool for quick, low-curation creation and solution of kinetic metabolic models. Previous versions of the DSAB have been successfully used to reconstruct and analyze large dynamic networks comprising hundreds of components. This paper presents updates included in Version 2.0 of the DSAB and the model solver: the inclusion of easy-to-use user interfaces for performing model construction, the inclusion of component pulse-feeding, and the introduction of regulatory inhibition kinetics. This study also describes the validation processes currently being carried out to beta-test the expansions to the DSAB; these tests utilize the purine metabolism pathways of Mycoplasm genitalium and Escherichia coli. Additionally, we have also incorporated inhibition schemes for the most-understood (catalyzed) reactions in our kinetic model of of the well-characterized Mus Musculus (including 15 principal metabolic pathways). Cascading factorial and composite factorial experimental designs were then used to explore the lone effects and interaction effects of enzyme expression levels, focusing on enzymes within the citric acid, glycolytic, and glutamate pathways. Our simulations suggest favorable interactions between the down-regulation of lactate dehydrogenase, iso-citrate dehydrogenase, and a-ketaglutarate dehydrogenase and several lesser-known enzymes might be exploited to reduce lactate and ammonia generation without compromising cellular growth.