Developing modern tools for metabolic engineering in a non-traditional host : fine-tuned protein production in Methanosarcina acetivorans C2A
Abstract/Contents
- Abstract
- New energy solutions are needed that are carbon independent or carbon neutral to curb the effects of global warming caused by burning fossil fuels to carbon dioxide. One carbon neutral energy technology that is gaining interest involves C1 fermentations using anaerobic microorganisms. In this process, hydrogen or carbon monoxide produced from clean, renewable technologies is mixed with carbon dioxide and transformed by anaerobic microorganisms into multi-carbon products, such as acetate and butanol. This process has the advantage to operate at high product specificity and yield at ambient temperature and pressures, which reduces the energy requirements for the process. In addition, there is opportunity for improved product yield and for tailored end product selection using metabolic engineering. However, metabolic engineering in these hosts has been challenging due to the lack of genetic tools available. This work developed tools for fine-tuned protein production that can be used for metabolic engineering in Methanosarcina acetivorans C2A, an Archaea capable of respiring carbon monoxide to produce acetate. First, a wide range of protein production was demonstrated through manipulation of basal transcription and translation regulation. In addition, the protein production range was further expanded by identifying new regulatory sequences in the 5' untranslated region. Finally, a CRIPSRi system was introduced for tunable repression of any gene in the genome. While the major protein components of the CRIPSRi system were functional in M. acetivorans C2A, further optimization is required to apply this system for metabolic engineering applications. The tools developed in this work for fine-tuned protein production will allow for the construction of more complex genetic circuits in M. acetivorans C2A and will be useful for metabolic engineering.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2017 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Karim, Ann |
|---|---|
| Associated with | Stanford University, Civil & Environmental Engineering Department. |
| Primary advisor | Spormann, Alfred M |
| Thesis advisor | Spormann, Alfred M |
| Thesis advisor | Sattely, Elizabeth |
| Thesis advisor | Welander, Paula |
| Advisor | Sattely, Elizabeth |
| Advisor | Welander, Paula |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Ann Karim. |
|---|---|
| Note | Submitted to the Department of Civil and Environmental Engineering. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2017. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2017 by Ann Amanda Karim
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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