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Permanent link (DOI): https://doi.org/10.7939/R3PN8XP56

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Metabolic Engineering of Saccharomyces cerevisiae for the Production of Isopentenol, a Valuable Biochemical and Potential Biofuel Open Access

Descriptions

Other title
Subject/Keyword
isopentenol
yeast
biofuel
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Pham, Diana Phuong Hong
Supervisor and department
Stuart, David (Biochemistry)
Examining committee member and department
Weiner, Joel (Biochemistry)
Bressler, David (Agricultural, Food & Nutritional Science)
Department
Department of Biochemistry
Specialization

Date accepted
2015-08-27T13:55:21Z
Graduation date
2015-11
Degree
Master of Science
Degree level
Master's
Abstract
The rising concern about limited fossil fuel resources and the environmental factors associated with it has motivated the development of alternative fuels. Ethanol is currently the primary substitute to gasoline, however the intrinsic properties of ethanol have limited its ability to fully replace petroleum based gasoline. Microbial production of higher chain alcohols is currently being explored to circumvent the problems associated with ethanol. Isoprenol is a 5-carbon alcohol that has a higher energy density and a lower water affinity when compared to ethanol. These characteristics allow isoprenol to fit well with existing fuel infrastructure. However, reported yields of isoprenol remain low and the process thus requires further optimization to increase production. Microbial production of isoprenol has been limited to synthetic pathways introduced into Escherichia coli. The approach in this study is to utilize the endogenous, Saccharomyces cerevisiae’s isoprenoid pathway and apply metabolic engineering approaches to manipulate gene expression and protein stability to increase flux through the pathway, maximizing isopentenyl pyrophosphate production. Isopentenyl pyrophosphate is an intermediate in the isoprenoid pathway that can be dephosphorylated to produce isoprenol. NudF, an exogenous phosphatase, with potential activity towards isopentenyl pyrophosphate, was expressed in Saccharomyces cerevisiae to produce isoprenol. Optimization of the isoprenoid pathway in Saccharomyces cerevisiae and expression of NudF did not lead to isoprenol production but did yield production of isopentyl alcohol. This work emphasizes the existence of inherent limitations when introducing novel processes into the cell.
Language
English
DOI
doi:10.7939/R3PN8XP56
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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