2008 Annual Report
Abstract
Our long-term goal is to use tools from molecular biology to engineer multi-enzyme metabolic complexes, mimicking the physical forms ubiquitous in nature. The direct coupling between sequential enzymatic reactions, through either static or dynamic interactions, offers the promise of eliminating these production barriers as it reduces the distance between enzyme active sites and favors sequential reactions over diffusion into the bulk. Therefore, the objective of these studies is to engineer synthetic metabolic complexes by exploiting the assembly mechanisms of natural systems to spatially organize enzymes that participate in sequential reaction steps. We expect that by developing a generic set of tools to co-localize metabolic pathways, we will overcome traditional bottlenecks that limit the commercial viability of microbial factories. We have proposed the following specific aims: (1) To demonstrate efficient multi-protein assembly in bacterial cells. To enable the intentional engineering of metabolic enzymes into functional metabolic complexes, we will explore a variety of novel methods for in vivo enzyme assembly; (2) To Assemble functional metabolic complexes. We will utilize the intracellular assembly/cross-linking methodologies from Aim 1 to create synthetic metabolic complexes in bacteria that are capable of efficient metabolic conversions via fermentation of renewable resources. We have chosen as a model system the microbial synthesis of propylene glycol (1,2-propanediol or 1,2-PD); (3) To enable combinatorial engineering of metabolic complexes via metabolite sensors. We will engineer a collection of intracellular switches that are capable of dynamically responding to intracellular metabolites (e.g., 1,2-PD) over a broad concentration range.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2008
- Accession Number
- ADA573190
Entities
People
- Matthew P. DeLisa
Organizations
- Cornell University