From Metabolites to Flux: Understanding Cellular Metabolism

Author

Cale Thornton, Boise State UniversityFollow

Publication Date

5-2024

Date of Final Oral Examination (Defense)

3-1-2024

Type of Culminating Activity

Thesis

Degree Title

Master of Science in Chemistry and Biochemistry

Department Filter

Chemistry

Department

Chemistry and Biochemistry

Supervisory Committee Chair

Owen McDougal, Ph.D.

Supervisory Committee Member

Lisa Warner, Ph.D.

Supervisory Committee Member

Joseph Dumais, Ph.D.

Abstract

Metabolomics is a well-established field with the first published mention of the metabolome in 1998 with numerous applications encompassing food science, microbiology, forensics, plant biology, and many others. In this thesis, I will describe how we utilized Nuclear Magnetic Resonance (NMR) spectroscopy to aid in our understanding of topics ranging from method development, biomass conversion, to food science. In the first topic, we will probe the challenges and successes in understanding how to monitor the real-time metabolism of living cells by NMR. Cells larger can ca. 0.2 μM in diameter tend to settle out of suspension within minutes. Pluronic F-127 is a biocompatible hydrogel that we tested as a cellular suspension agent for in-cell NMR. This hydrogel keeps cells suspended within an NMR tube preventing cell settling, while still allowing for cell homeostasis and metabolism. This hydrogel has the potential to expand the utilization of in-cell NMR experimentation to a wider variety of cell types and sizes. Next, we explored the metabolism of Clostridium thermocellum, an anaerobic bacteria used in biomass conversion. Clostridium thermocellum is an efficient degrader of cellulose, converting plant waste into ethanol. and also has an additional CO₂ fixation pathway. This metabolism of CO₂ was relatively uncharacterized. In-cell NMR analysis identified major downstream metabolites of CO₂ consumption as being lactate and formate. The final aspect of this research is utilizing metabolomic techniques is the comparison of wines prepared traditionally and with pulsed electric field processing. Pulsed electric field processing was found to produce a final wine product spectroscopically indistinguishable as wines that were prepared traditionally. This thesis hopes to highlight some of the many types of metabolomic analysis possible.

DOI

https://doi.org/10.18122/td.2225.boisestate

Recommended Citation

Thornton, Cale, "From Metabolites to Flux: Understanding Cellular Metabolism" (2024). Boise State University Theses and Dissertations. 2225.
https://doi.org/10.18122/td.2225.boisestate