MURI: Translating Biochemical Pathways to Non-Cellular Environment

MURI: Translating Biochemical Pathways to Non-Cellular Environment Translating Biochemical Pathways to Non-Cellular Environment Summary of our technical approach: Biochemical pathways that convert mass or energy govern the cellular activities of all organisms. The function of these pathways is highly dependent on the precise arrangement of participating actuators, typically proteins. The ability to design and control pathways beyond natures repertoire would enable enzyme-catalyzed production of novel molecules and energy conversion optimized for ambient and extreme environments. One promising route towards achieving this goal is to develop approaches that translate biochemical reaction pathways to non-cellular environments, where they can be easily manipulated. With our recent advances in complex 1D, 2D and 3D DNA nanostructure engineering, it now has become feasible to assemble multi-component pathways on spatially addressable molecular DNA pegboards without compromising component functionality. To realize this vision, we have established a multidisciplinary team with expertise in DNA nanotechnology, computational biology and biophysics, protein chemistry and bioconjugation, photo-biochemistry, porous material synthesis and single molecule biophysics to design, model, construct and characterize DNA templated biochemical pathways that convert either mass or energy, and to realize their optimal system functionality in a non-cellular environment. Three main thrusts will be pursued: i.) Multi-dimensional scaffolding, directed substrate channeling and programmable compartmentalization will be tested to control and facilitate substrate diffusion in multi-enzyme pathways; ii.) selected bioelectroactive pathways will be interfaced with an external, conductive material for possible device applications; and iii.) uniform light absorption, efficient energy conversion and improved photostability will be engineered for the construction of artificial light harvesting complexes. Computational modeling and singlemolecule and bulk characterization will be employed to determine the parameters most critical to assembling these functional biochemical pathways and iteratively feed back into the design. Anticipated outcome and potential impact on DoD capabilities: Our proposed scope of work develops nanoscale devices based on important biological systems (enzyme cascades and photosynthesis) to advance our understanding of the complexities of translating biology at the nanoscale. DNA directed assembly allows us to control and exploit interactions between synthetic and naturally occurring materials for low-cost synthesis and templating of designed nanostructures. Our architectures will be used to enhance local diffusion behavior, reaction kinetics, and the optical and electrical properties within the proposed devices, behavior that can be extended to other systems of interest to the DoD. In addition, the computational modeling and single molecule characterization that we intend to perform will increase our understanding of the properties of nanomaterials, including interface interactions, enabling the design of nanostructured materials with targeted properties. In line with DoD goals, the proposed research will increase our understanding of nanoscale, enzymatic and energy transfer processes and aid in the development of robust strategies for the synthesis, characterization, and assembly of synthetic biochemical pathways for application in revolutionary catalysis, high-efficiency photoenergy generation, energy storage devices, sensors, or completely new capabilities that enhance war fighter and battle system capabilities. Supplement: MURI: Translating Biochemical Pathways to Non-Cellular Environment