Date of Award

Fall 11-2018

Document Type

Thesis

Degree Name

Master of Science in Chemical Engineering (MSChE)

Department

Chemical Engineering

First Advisor

Reizman, Irene

Second Advisor

Goulet, David

Third Advisor

Coppinger, J. Peter

Abstract

Optogenetics has gained increasing attention for enabling reversible and non-invasive control in biochemical engineering. The purpose of this project is to investigate the application of a genetically engineered light-switchable system, Cph8-OmpR, for controlling the accumulation of the useful central metabolite glucose-6-phosphate (G6P) in Escherichia coli. This could be accomplished by linking expression of an adaptor protein, SspB, which increases enzyme phosphofructokinase (Pfk) degradation, with different light intensities. Pfk is the enzyme that catalyzes the conversion of fructose-6-phosphate (F6P), an isomer of G6P from an equilibrium reaction, to fructose 1,6-biphosphate for downstream metabolism. This step is considered the major control point of G6P accumulation in the experimental bacterial strain. A mathematical model was successfully established for this system and predicted that lower red-light exposure would result in higher intracellular G6P concentration. A construct for experimental tests was successfully made, but more work on plasmid construction and strain modification is required for future experiments.

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