Retrobiosynthesis of D-glucaric acid in a metabolically engineered strain of Escherichia coli

Synthetic biology is an evolving field that emphasizes synthesis more than observation which has been and is the scientific method for traditional biology. With powerful synthetic tools, synthetic biologists seek to reproduce natural behaviors (and eventually to create artificial life) from unnatural molecules or try to construct unnatural systems from interchangeable parts. Accompanied with this recent movement, metabolic engineers started to utilize these interchangeable parts (enzymes in this case) to create novel pathways. In addition, engineering biological parts including enzymes, promoters, and protein-protein interaction domains has led to productivity improvement. However, understanding behaviors of a synthetic pathway in an engineered chassis is still a daunting task, requiring global optimization. As the first step to understand pathway design rules and behaviors of synthetic pathways, a synthetic pathway for the production of D-glucaric acid has been designed and constructed in E. coli. To this end, three disparate enzymes were recruited from three different organisms, and the system perturbed by this introduction of heterologous genes was analyzed. Limiting flux through the pathway is the second recombinant step, catalyzed by myo-inositol oxygenase (MIOX), whose activity is strongly influenced by the concentration of the myo-inositol substrate. To increase the effective concentration of myo-inositol, synthetic scaffold devices were built from protein-protein interaction domains to co-recruit all three pathway enzymes in a designable complex.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Chemical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2010

Author dc:creator

• Moon, Tae Seok

Advisor dc:contributor.advisor

• Kristala Jones Prather.

• Chemical Engineering.

Statement dc:rights

• M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

http://hdl.handle.net/1721.1/57868

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/57868