Abstract
Space exploration has long been a source of inspiration, challenging scientists and engineers to find innovative solutions to various problems. One of the current focuses in space exploration is to send humans to Mars. However, the challenge of transporting materials to Mars and the need for waste management processes are two major obstacles for these long-duration missions.
To address these two challenges a process called Astroplastic was developed that produces polyhydroxybutyrate (PHB) from solid human waste, which can be used to 3D print useful items for astronauts. PHB granules are naturally produced by bacteria such as Ralstonia eutropha and Pseudomonas aeruginosa for carbon and energy storage. The phaJ, phaC, and phaCBA genes were cloned from these native PHB-producing bacteria into Escherichia coli. These genes code for enzymes that aid in PHB production by converting products of glycolysis and β-oxidation pathways, such as acetyl-CoA and enoyl-CoA, into PHB. To ensure a continuous PHB production system and to eliminate the need for cell lysis to extract PHB, recombinant E. coli was engineered to use the genes in its natural type I secretion system to secrete PHB. The C-terminal of the HlyA secretion tag was fused to phasin (PhaP), a protein originally from R. eutropha. Phasin-HlyA electrostatically binds PHB granules and transports them outside of the cell.
In addition to genetically engineering bacteria, a concept for start-to-finish PHB production process was designed. Integrating expert feedback and experimental results, conditions for each step of the process including the collection and storage of waste, volatile fatty acid (VFA) fermentation, VFA extraction, PHB fermentation, and PHB extraction were optimized. The optimized system will provide a sustainable and continuous PHB production system, which will address the problems of transportation costs and waste management for future space missions.
Financial Disclosure Mindfuel Science Alberta Foundation Genome Alberta GenScript Polyferm Canada GeekStarter Alberta Integrated DNA Technologies University of Calgary University of Calgary Cumming School of Medicine University of Calgary Bachelor of Sciences University of Calgary Schulich School of Engineering University of Calgary O’Brien Centre for the Bachelor of Health Sciences City of Calgary Alberta Innovates The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests The authors have declared that no competing interests exist.
Ethics Statement N/A
Data Availability All data are freely available without restriction.
Author Contributions
Conceptualization: KS XC AK PG MAN HWC SI ZW AI MO JG PL LB BS RCV MAP Methodology: AK KS XC SI MAN HWC JG AI PG ZW LB PL MO BS Software: MO Formal Analysis: AK MAN AI Investigation: KS XC PG SI AK HWC MAN ZW LB JG PL MO RCV DF BS Resources: MAP RCV AK DF Writing Original Draft Preparation: XC SI AK ZW PG KS MAN HWC JG LB Writing – Review & Editing: XC AK SI ZW MAP LB HWC Visualization: TL MO XC AK HWC Supervision: MAP RCV DF Project Administration: MAP RCV DF AK XC SI KS Funding Acquisition: MAP RCV AK KS XC PG SI MO PL ZW LB MAN HWC BS JG