Engineering in vitro photobiological hydrogen production
Abstract/Contents
- Abstract
- Hydrogen is an important industrial chemical and attractive next-generation fuel. Current methods for its production rely on steam reformation of fossil fuels which liberates greater than 1% of the annual global carbon dioxide emissions; therefore, there is a need for sustainable hydrogen production processes. Nature has provided a biological catalyst for hydrogen production in [FeFe] hydrogenases. These complex enzymes catalyze the reaction which combines electrons and protons to generate hydrogen. Our long term goal is to integrate a non-native [FeFe] hydrogenase into the photosynthetic electron transport chain of a cyanobacterium to couple photosynthetic water-splitting with hydrogen production. One challenge to implementing this pathway is that heterologous hydrogenases interact inefficiently with the photosynthetic machinery and electrons will be lost to native, competing pathways in the cyanobacteria. Engineering the hydrogenase and other pathway components in vivo is challenging due to the limited tools available for genetic engineering these photosynthetic organisms. We developed an improved in vitro pathway using Photosystem I complexes isolated from a cyanobacterium to mimic in vivo electron flow to the hydrogenase or the main competing pathway, the production of NADPH. This system provides fine-tuned control over the reaction components and environment including concentrations of proteins in the pathways to study and control electron flux. We demonstrate the use of a non-native ferredoxin to facilitate electron transfer between the photosystem and hydrogenase. Finally, we will describe coupling our photosystem-pathway with a high-throughput hydrogen sensor to assess hydrogen production rates in microtiter plates. This approach can be extended to the high-throughput screening of in vitro synthesized enzyme or electron carrier libraries to develop components that support high photobiological hydrogen production rates.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2015 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Shiigi, Stacey Akie |
|---|---|
| Associated with | Stanford University, Department of Bioengineering. |
| Primary advisor | Swartz, James R |
| Thesis advisor | Swartz, James R |
| Thesis advisor | Cochran, Jennifer R |
| Thesis advisor | Smolke, Christina D |
| Advisor | Cochran, Jennifer R |
| Advisor | Smolke, Christina D |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Stacey Akie Shiigi. |
|---|---|
| Note | Submitted to the Department of Bioengineering. |
| Thesis | Thesis (Ph.D.)--Stanford University, 2015. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2015 by Stacey Akie Shiigi
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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