Date Presented

Spring 5-18-2024

Document Type

Thesis

First Advisor

Dr. Samrat Thapa

Second Advisor

Dr. Jamie Brooks

Third Advisor

Dr. Jason Crumpton

Abstract

Ethylene is the organic compound that serves as the raw material for polyethylene and other chemical polymers used in the manufacturing of plastic products. Traditional methods used to obtain ethylene are associated with significant carbon emissions that are harmful to the environment. Despite these negative environmental impacts, the utility of ethylene in commercial and medical industries has led researchers to investigate potential pathways for ethylene biosynthesis in recent decades. Previous studies have identified an ethylene-forming enzyme (EFE) that is involved in ethylene biosynthesis in many microorganisms. Researchers have explored biochemical properties of EFE and strategies for maximizing enzyme expression and ethylene yield in recombinant organisms expressing the efe gene; however, it remains unclear whether ethylene biosynthesis involving EFE can produce the high ethylene yields necessary for commercial and medical use. Research conducted in this project investigated ethylene biosynthesis as a potential alternative to conventional methods used to obtain ethylene. Experiments were conducted to analyze the structure and kinetic properties of EFE from Pseudomonas syringae. Research focused on enzyme isolation and purification from recombinant Escherichia coli engineered to express the efe gene. Nuclear magnetic resonance (NMR) spectroscopy was then used to analyze the kinetic properties of EFE. Future research using high-performance liquid chromatography (HPLC) to analyze the kinetic properties of EFE and indirectly measure EFE ethylene production is necessary to further evaluate ethylene biosynthesis via the ethylene-forming enzyme as an alternative to traditional ethylene production methods.

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