Characterizing uronate dehydrogenase from P. syringae as a step towards expressing a retro-biosynthetically derived glucaric acid pathway in E. coli

Retro-biosynthesis is a novel pathway design method that integrates biocatalysis (in particular, functional group specificity and reactivity of enzymes) and organic chemistry. Using this method, several novel pathways have been designed in this laboratory for the microbial production of glucaric acid. In addition to design, efforts have been made in order to express these pathways inside E. col. For the production of glucaric acid in E. coil, toxicity of this organic acid as well as the uptake efficiency of glucose, the starting substrate, when in the presence of glucaric acid was assessed. Furthermore, a benchmark pathway for glucaric acid production that utilizes known reaction steps from different source organisms was designed. The design and expression of this pathway will be compared to the retro-biosynthetic pathway. The ease of design and expression will be analyzed in order to confirm the validity of retro-biosynthesis. Toxicity results showed that glucaric acid concentrations, up to 40 mM, do not affect growth of E. coli DH1OB cells. Furthermore, cells take up equal amounts of glucose even when glucaric acid is available, indicating that glucose is preferentially utilized by cells and that glucaric acid, once produced by cells, would not affect further production.

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Chemical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2006

Author dc:creator

• Iranpour, Pooya

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/1721.1/35135

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/35135