System analysis and metabolic engineering of biocatalytic reaction networks : application to indene bioconversion

Metabolic engineering is a science concerned with the improvement of cellular properties through the application of recombinant DNA technology. To this point, the field has frequently been limited to methodological issues pertaining to gene introduction, expression and control by way of molecular biological techniques. While such methods are critical for the implementation of strategies for metabolic pathway design and modification, an equally critical activity is the quantitative analysis of metabolic networks and the evaluation of phenotype/genotype relationships. A prominent new opportunity to extend the scope of metabolic engineering has recently emerged to analyze and engineer biocatalytic microorganisms capable of catalyzing stereospecific transformations useful in chiral pharmaceutical manufacturing. This thesis formulates and applies a general framework for the optimization of uncharacterized bioconversion strains (those biocatalysts with little a priori genetic or metabolic data available) comprising five essential steps: (i) Establishment of an experimental system for strain selection and metabolic network analysis, (ii) definition of the bioconversion network and quantification of network fluxes, (iii) target identification, (iv) flux redistribution, and (v) analysis of the modified bioconversion strain to identify areas for further development. In the case described here, a systematic evaluation of the physiology of the soil bacterium Rhodococcus, and analysis of the relative fluxes of an indene bioconversion network,

Department dc:contributor.department

Massachusetts Institute of Technology. Dept. of Chemical Engineering.

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2002

Author dc:creator

• Stafford, Daniel Edward, 1975-

Advisor dc:contributor.advisor

• Gregory Stephanopoulos.

• 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/1721.1/8271

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/8271