The impact of organizational strategy on performance of in vitro biosynthesis reactions

Project: Research project

Project Details

Description

There is a need to be able to synthesize biomolecules and sense chemical environments in resource-limited and far forward areas for the protection of the soldier. Synthetic biology offers an emerging approach to do so, but issues remain. One primary issue facing biological engineering efforts is to control precisely when and where biochemical reactions take place to achieve desired products at relevant yields. Nature makes use of compartmentalization strategies, including the bacterial organelles known as microcompartments (MCPs), to accomplish this spatial control and achieve outcomes unattainable without such organization. A particularly exciting area is to repurpose these natural microcompartments to direct outcomes for engineered biological systems. To enhance metabolic pathways and sensing systems, several questions must be answered related to the primary function and utility of the compartmentalization systems. This proposal seeks to use the propanediol utilization MCP system with two model pathways, the mevalonate pathway and the violacein pathway, to study the following:
Aim 1: Evaluate quantitatively the impact of compartmentalization on yield, and product profiles for the model metabolic pathways;
Aim 2: Determine whether a private cofactor pool affects pathway performance for the model metabolic pathways; and
Aim 3: Develop a scaffold system that enables comparison of pathway performance between colocalized, encapsulated, and free enzymes using identically modified pathway enzymes.
Uniquely, the experiments with both pathways will be examined in the in vitro context of cell-free lysates using a cell-free metabolic engineering platform to reduce the confounding factors typical of living systems. The studies, if successful, will experimentally determine the biochemical benefits of spatial organization of metabolic pathways and enable the design of spatiotemporal organization in bioengineered systems.
StatusActive
Effective start/end date5/15/195/14/22

Funding

  • Army Research Office (W911NF-19-1-0298)

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