COL. CHRIS HADFIELD

Profession: Former Astronaut
Canadian Space Agency
We received advice from Col. Chris Hadfield (and his dog, Albert) regarding the feasibility of our project on Mars, its integration into current endeavours and research into space exploration, and what we could do to improve our project's design to satisfy these key concerns. Chris reminded us that the "prime directive" to any space mission is the safety and survival of the crew, and this was thoroughly reviewed and assessed in the development of our system. In addition, he provided us with insight regarding important policies and regulations to consider as we moved forward with our project. This included treaties and agreements released by the United Nations Committee on the Peaceful Uses of Outer Space (COPUOS), and emphasized the containment of foreign biological systems (like engineered bacteria) to new planets.

CHANCELLOR DR. ROBERT THIRSK

Profession: Former Astronaut
Canadian Space Agency
The team met with the University of Calgary Chancellor Robert Thirsk and discussed the demand for on-site manufacturing and recycling of solid human waste on Mars. Dr. Thirsk provided us with advice regarding ways to improve our project's design while considering key issues commonly dealt with during space missions. One of the key issues that we discussed were In-Flight Maintenance Requirements, keeping in mind that any machine that is brought up and used in space should be one that can be easily repaired by crew members. This was taken into consideration by our Process Development team in designing the key components of our PHB-producing system.

DR. MATTHEW BAMSEY

Profession: Chief Systems Engineer
German Aerospace Center
Dr. Bamsey provided us with documents concerning Equivalent System Mass, published by NASA, which provide a universal means to calculate the energy required and efficiency of items brought to space. We used this information to compare different life support systems as we considered how to improve our project's design. Matt also directed us to Dr. Nicole Buckley of the Canadian Space Agency to gather more information to support the biological side of our project.

DR. NICOLE BUCKLEY

Profession: Microbiologist
Canadian Space Agency
Dr. Buckley helped us by identifying issues that we would need to resolve in the future for space travel. These issues include the potential of viral shedding from human feces, which could result in increased disease prevalence onboard the spacecraft as viruses may spread to fellow astronauts. As such, the team is currently considering UV treatment to assist in sterilizing human waste after it has been left to ferment with natural gut flora. She also pointed out the potential for abnormal or diminished bacterial growth due to a microgravity environment, and helped us decide to employ our system only on Mars to avoid this issue. Dr. Buckley also identified the need to bring all of our cell maintenance materials on board the spacecraft, pushing us towards our less material-intensive PHB secretion recovery system as opposed to a lysis system.

DR. PASCAL LEE

Profession: Principal Investigator of the Haughton-Mars Project
NASA Ames Research Center

DR. DEREK THOMAS

Profession: Senior Materials Scientist
Made in Space, Inc.
We discussed our project with Derek Thomas from Made In Space, Inc., a company that developed the 3D printer currently in use on the International Space Station. Derek commented on the current uses of the 3D printer on the ISS, which include producing tools and other small items to assist with everyday operations. Having the capability to manufacture items on hand will be especially important on Mars because longer mission duration and distance will make it challenging to transport necessary materials. Derek also identified safety as one of the most important factors to consider when designing systems for space applications. When considering the use of PHB on Mars, Derek advised analyzing flammability, gassing characteristics, and other physical properties to identify the most suitable applications for our product. He suggested considering post-processing to obtain any desired characteristics that are lacking in pure PHB. Regarding materials that can be used to build the system itself, his advice was to first identify the desired requirements and operating conditions. Lastly, Derek commented on the use of Selective Laser Sintering (SLS) 3D printer. Although he does not foresee major concerns with the use of SLS 3D printer on Mars, controlling the powder at Mars' gravity might be more challenging. Additionally, exposure to lower atmospheric pressure has the potential to cause problems.

DR. BRUCE RAMSAY

Profession: Founder and CTO
Polyferm
He encouraged us to pursue a lysis or secretion system due to its novelty, however cautioned that the final separation process is the most inefficient. Focusing on the increasing the final yield would be important from a marketing perspective.Currently there is not a high demand for bioplastics due to the cost of their porduction. Most medium chain polymers are used by businesses for experimentation, but not for large scale production. This information prompted our team to look towards which applications might benefit most from being able to produce plastic. We immediately thought of space, as the high cost of shipping materials would make a process that could produce plastic on site an attractive proposition. Furthermore, the ISS already uses 3D Printers so our product could be used to create useful materials for astronauts.

MARKO MARKICEVIC

Profession: Customer Service Liaison, Lab Operations for Water Quality Services
Calgary Wastewater Labs
We spoke with Marko Markicevic from the City of Calgary Water Quality Services lab, who do most of the water quality testing on samples collected within the City’s wastewater treatment plant. We learned that VFAs are currently analyzed primarily in primary effluence and fermenter supernatants of the plants, however only the short chained VFAs are monitored. Based on this information we decided that adding an additional VFA Fermentation step in our process would be beneficial to increasing PHB yield. Marko also encouraged us to consider the upfront costs of incorporating our system into existing wastewater facilities, and whether using pure cultures is the best decision. He mentioned that the wastewater treatment industry already uses methods for production of value added products (methane, fertilizers etc.), and that those process are much less equipment and "cleaning" intensive than the PHB production process. This made it less likely that our project would survive on the market which encouraged us to look for other applications.

DARINA KUZMA

Profession: ACWA Analytical Lab Manager
ACWA
We spoke with Darina Kuzma from ACWA (Advancing Canadian Wastewater Assets). Their research focuses on water that has already gone through the sanitation process and finding ways of eliminating particles that are too small for regular methods to detect (such as over the counter medication and other drugs). She mentioned that if our secreted PHB particles are too small, we would need to consider carefully the environmental safety of our process if these particles left the containment of a wastewater treatment plant and actually entered water streams and rivers. This prompted the engineering team to consider possible extraction methods, such as coagulation, that could mitigate this risk.

CAROL NELSON

Profession: Waste Management Specialist
Alberta Environment and Parks
Carol Nelson told us that leachate from industrial landfills have little to no organic content, which is not applicable to our project; however, the leachate from municipal landfills enters wastewater treatment plants and so would be safe to use as a feedstock. Currently, leachates with hazardous chemicals and metals enter deep well injections which are very cheap. If we applied our PHB production to landfills, they would have to compete with deep well injections, which due to the current low demand of PHB plastics might not be worth it from an economic perspective. Because of this information, we chose not to pursue a landfill leachate application at this time, but look forward to the possibilities of applying our project to this field in the future.

DR. SUI LAM WONG

Profession: Professor, Microbiology
University of Calgary
Dr. Wong provided insight on antibiotic free containment, ultimately leading us to the auxotrophy system that we discuss in our Safety page. He also informed us on how to conduct protein expression assays through the use of an inducible promoter system so we could measure the activity of exogenous genes. He provided us with E. coli DH5𝛼 which he instructed us to use for the transformation and propagation of our ligated plasmids. He then provided us with E. coli BL21(DE3), a strain used specifically for protein expression. Lastly, he donated pET29B(+) to our team, a vector with an IPTG-inducible T7 promoter that was used in correspondence with E. coli BL21(DE3). With Dr. Wong's assistance, we were able to construct our plasmids and express recombinant proteins in a quantifiable manner.

DR. ELKE LOHMEIER-VOGEL

Profession: Senior Instructor, Biochemistry
University of Calgary
Dr. L-V was a regular attendee of our lab meetings wherein she provided us with insight regarding the alteration of biochemical pathways in E. coli. She directed us to consider the flux of incoming nutrients versus the output of bioplastic, as the metabolic pathways we employed to produce PHB are in direct competition with cell energy production. This allowed for the team to begin modelling flux balance analysis and kinetics to identify possible bottlenecks in our system, which has the potential to later allow us to optimize PHB production by genetic knockout or providing our bacteria with an alternate nutrient medium.

DR. JUSTIN MACCALLUM

Profession: Assistant Professor, TIER II CRC Biomolecular S & D, Physical & Theoretical Chemistry
University of Calgary
In our meeting with Dr. MacCallum we discussed about our plans for developing a model for our project. The two models that our modelling team considered were flux balance analysis and kinetic modelling. Dr. MacCallum told us about how we could incorporate Type I secretion system into our flux balance model along with the PHB synthesis pathways. Thus, this helped us to work toward a model that could closely simulate the lab experiments. He also referred us to Dr. Lewis whose team worked on flux balance analysis.

DR. IAN LEWIS

Profession: Assistant Professor, Metabolomics and Biochemistry
University of Calgary
In our meeting with Dr. Lewis and his team of postdocs, we discussed the flux balance model in details. We were told about how flux balance will be helpful for informing our project as it will tell us the optimal pathway that E. coli will use after inserting our constructs. Furthermore, the team was told about how flux balance analysis could progress into flux variability analysis, which will help us to modify parameters in the model and analyze the change in PHB production.

DR. JASON DE KONInG

Profession: Assistant Professor, Bioinformatics, Biochemistry and Molecular Biology
University of Calgary
We had our very first meeting regarding modelling for our project with Dr. De Koning. In our meeting we were told about codon optimization and how it could be used to improve our constructs. Thus, the team decided to codon optimize all the biobricks submitted to the registry. We were able to improve a previous part. The team also discussed the potential models for the project with Dr. De Koning. He provided us with resources and software for finding parameters for our kinetic model.

DR. NASHAAT NASSAR

Profession: Associate Professor, Chemical and Petroleum Engineering
University of Calgary

DR. SAURABH JYOTI SARMA

Profession: Postdoctoral Scholar
University of Calgary