Date of Graduation
Master of Science, Biology
Gluconobacter oxydans, fluorescent reporter, inducible promoter, fusion protein, genetic tools
The acetic acid bacterium Gluconobacter oxydans is an industrially valuable microorganism, particularly in the production of acetic acid, D-gluconic acid, ketogluconic acids, dihydroxyacetone, and precursors for the antidiabetic drug miglitol. Despite its importance in industry, there is still much to be learned about G. oxydans and its many uncharacterized enzymes. Additionally, genetic engineering holds the possibility of improving current yields. However, these goals are limited, largely due to a lack of molecular tools suitable for working with the bacterium. The current molecular toolkit for G. oxydans specifically lacks an efficient screening system for positive clones and a system for regulatable gene expression. Therefore, two sets of fluorescent protein-based reporter systems were designed for screening and protein purification along with two sets of inducible promoter systems, the Tet and Lux systems, for the regulation of gene expression. Although, the fluorescent reporter systems were unreliable as a gene expression screening tool, the Lux inducible promoter system had reliable and strong induction of gene expression, and is a promising system to use for metabolic engineering and regulatable gene expression in G. oxydans.
© Melissa A. Schoeben
Schoeben, Melissa A., "Genetic Tools to Allow Efficient Gene and Protein Characterization of the Industrially Important Bacterium Gluconobacter Oxydans" (2017). MSU Graduate Theses. 3117.