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Investigation of the Saccharomyces cerevisiae peroxisome and beta-oxidation

Spädt, Erik (2025) MOBN02 20242
Degree Projects in Molecular Biology
Popular Abstract
Saccharomyces cerevisiae: Genome editing and investigation of the peroxisome & beta-oxidation

For thousands of years humans have used yeast to brew beer, bake bread and make wine. Nowadays, these microorganisms can be used to produce various chemicals and proteins. One interesting compound is capsaicin, found in the chili pepper plant and commonly used in food, but lesser-known for its application in pain treatment. However, extracting it from plants is dependent on the climate and specific environmental conditions. Chemical synthesis is possible, but environmentally unfriendly. Therefore, the idea is to produce it in yeast. One problem is the toxicity of capsaicin to cells, but how do plants deal with it? They compartmentalize their... (More)
Saccharomyces cerevisiae: Genome editing and investigation of the peroxisome & beta-oxidation

For thousands of years humans have used yeast to brew beer, bake bread and make wine. Nowadays, these microorganisms can be used to produce various chemicals and proteins. One interesting compound is capsaicin, found in the chili pepper plant and commonly used in food, but lesser-known for its application in pain treatment. However, extracting it from plants is dependent on the climate and specific environmental conditions. Chemical synthesis is possible, but environmentally unfriendly. Therefore, the idea is to produce it in yeast. One problem is the toxicity of capsaicin to cells, but how do plants deal with it? They compartmentalize their capsaicin production into specific tissues, so it does not affect other cells. Taking this as an example, it might be possible to transfer the production of capsaicin into yeast peroxisomes. To evaluate the potential of the peroxisome to harbor the capsaicin-producing machinery, genes affecting peroxisome structure and function were studied.

Experiments with gene deletions were performed to study their impact on yeast growth on oleic acid, but none of the tested genes seem to affect yeast growth much. Other experiments utilizing fluorescent proteins showed that the FOX3 / POT1 gene becomes active when yeast are grown on fatty acids, which might be important in the future for cell factory design. Lastly, a toolbox for genetic engineering of yeast beta-oxidation was evaluated for its functionality. It uses Cas9 – an enzyme which cuts DNA – and a guideRNA which leads Cas9 to the correct location. This guideRNA can be exchanged as desired and thus the genome can be edited multiple times and at different sites. This work shows the successful application of this method to delete the FOX1 gene from yeast and later re-integrate it back into the genome.

In the future, synthetic genes encoding enzymes with suitable activity and specificity can be used to replace the original yeast beta-oxidation genes. By affecting the fatty acid metabolism, the desired length of fatty acids can be reached, which is crucial to produce capsaicin.

Master’s degree project in Molecular biology, microbiology and biotechnology. 45 ECTS
Department of Biology, Lund University

Supervisor: Magnus Carlquist, Co-Supervisor: Jacob Miefalk
Division of BAM, Department of PLE, LTH, Lund University (Less)
Please use this url to cite or link to this publication:
author
Spädt, Erik
supervisor
organization
course
MOBN02 20242
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
9214816
date added to LUP
2025-11-03 14:11:05
date last changed
2025-11-03 14:11:05
@misc{9214816,
  author       = {{Spädt, Erik}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Investigation of the Saccharomyces cerevisiae peroxisome and beta-oxidation}},
  year         = {{2025}},
}