Description
- Abstract:
- There remains an unmet need for reliable access to pharmaceuticals on a multi-year crewed space exploration mission. Pharmaceuticals are set, conservatively, to expire within 2 years of manufacture. The viability of the limited quantity flown at launch (due to finite cargo space), is highly dependent on manufacturing lot, storage conditions, and exposure to space hazards, such as radiation and microgravity. Previously, the 2019 Brown-Stanford-Princeton International Genetically Engineered Machine (iGEM) Competition team presented the Astropharmacy, a system for the on-demand synthesis of peptide-based drugs with a Bacillus subtilis-based expression system, that aims to address the core challenges of obtaining pharmaceuticals in space. Here, I aimed to optimize the growth temperature and expression times of transgenic B. subtilis for teriparatide synthesis using both luminescent and chromogenic reporters to determine relative yield. Using a luminometer, luminescence was determined after 24 hours of growth at variable temperatures. A spectrophotometer was used to measure optical density (OD600) values of the luminescent bacteria after 24 hours of growth. A similar method was used to determine growth curves for B. subtilis spores expressing teriparatide-chromogenic marker constructs over 48 hours. The optical density of B. subtilis producing filgrastim- and teriparatide-luminescent marker peptides was greatest at 40 °C. However, growth at 20 °C produced significantly more teriparatide per milliliter of culture than at other temperatures. When incorporating a purple chromogenic marker in place of the luminescent marker, no color output was observed in our plated colonies. Understanding the growth patterns and expression of target peptides in transgenic B. subtilis is crucial for optimizing the overall Astropharmacy, which aims to increase accessibility to crucial drugs in space and in environments with unstable storage conditions, limited resources, and no opportunity for resupply. While the impetus for this work is human space exploration, our work can also be applied on Earth in any situation where such conditions may exist, such as in remote or rural regions.
- Notes:
- Thesis (Sc. M.)--Brown University, 2023
Citation
Bui, Cynthia Vivian,
"THE ASTROPHARMACY: ENGINEERING BACILLUS SUBTILIS FOR PEPTIDE-BASED DRUG PRODUCTION IN SPACE"
(2023).
Biology and Medicine Theses and Dissertations, Biotechnology.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:gkjjdpna/
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Biology and Medicine Theses and Dissertations
Theses and Dissertations for the Biology and Medicine department....