Office of Naval Research Annual Report
Abstract
Our overall objective is to facilitate the rational design of biosynthetic pathways for the production of unnatural compounds. We are developing tools and attempting to elucidate rules to establish a framework for retro-biosynthesis, the practice of rationally proposing a synthetic scheme for a target compound from one or more starting substrates, based only on enzyme-mediated transformations. We are especially focused on experimental realization of microbial synthesis, and the limits thereof, in contrast to theoretical predictions. A critical tool is protein engineering for altered substrate specificity, either through rational protein design or random mutagenesis (directed evolution). We have chosen glucaric acid as a model compound for pathway proposition and assembly in Escherichia coli. This compound is naturally-occurring in plants and mammals, but a microbial pathway has not been established. Specific objectives are to: (1) propose several pathways for glucaric acid synthesis, (2) select at least one pathway for experimental study, and (3) establish microbial synthesis of glucaric acid. A fourth objective arising from this work is the development of a database for the re-classification of enzyme activities based only on substrate and product functional groups. The database is designed to enable searches more amenable to enzyme selection for biosynthetic pathway proposition.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2008
- Accession Number
- ADA573189
Entities
People
- Kristala L. Jones-prather
Organizations
- Massachusetts Institute of Technology