Team:Calgary/HP/Gold Integrated

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Integrated Human Practices

Below is a detailed list of the experts we spoke to while we were developing our constructs and process, the feedback they gave us, and specific areas of our project where we integrated this information.

Mission Control

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 of 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 containment of foreign biological systems (like engineered bacteria) on 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. He also directed us to the NASA Life Support Baseline Values and Assumptions document containing a set of assumptions that can be used in the Equivalent System Mass analysis. Dr. Bamsey 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 biological issues that we would need to resolve for future 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 could 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 under the gravitational pull of 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
We spoke to Dr. Lee about the feasibility of our project and which design criteria we should keep in mind when developing our process. Based on his suggestion, we researched potential applications of solid human waste that NASA is considering so we could integrate our project with NASA's plans while identifying useful applications of our byproducts. Our findings can be found on the Applied Design and Products pages. Dr. Lee also highlighted the importance of considering energy availability and safety when developing our process. We considered energy availability by performing an Equivalent System Mass analysis and considering NASA's Life Support Baseline Values and Assumptions. During our meeting, we also discussed how space technologies are developed and tested.

DR. DEREK THOMAS

Profession: Senior Materials Scientist
Made in Space, Inc.
Made In Space, Inc. is a company that developed the 3D printer currently in use on the International Space Station. Dr. Thomas 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 on-hand manufacturing will be especially important on Mars because longer mission duration and distance will make it challenging to transport necessary materials. Dr. Thomas 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, he advised we analyze flammability, gassing characteristics, and other physical properties to identify the most suitable applications for our product. He also 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, Dr. Thomas commented on the use of Selective Laser Sintering (SLS) 3D printer, which can use powdered PHB for printing. Although he does not foresee major concerns with the use of SLS 3D printer on Mars, controlling the powder under Mars' gravity might present a challenge that we took into consideration.

DR. BRUCE RAMSAY

Profession: Founder and CTO
Polyferm
Dr. Ramsay provided us with insight to the current industrial processes of PHB production. While encouraged us to pursue a lysis or secretion system due to its novelty, he cautioned that the final separation step in traditional PHB production lines is the most inefficient one. Focusing on the increasing the final yield would be important to ensure our product produced a net profit from a marketing perspective. Presently, the demand for bioplastics is not high because of their costly production methods, particularly when cells are lysed with expensive solvents to harvest PHB. In addition, most medium chain polymers are used by clients for experimentation, and production is not required on a large scale. This information prompted our team to look towards which applications might benefit most from being able to produce our bioplastic. We immediately thought of space, as the high cost of shipping materials would make a process that could produce plastic on site an attractive alternative. Furthermore, the ISS already uses 3D printers, demonstrating how 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. The team there completes the majority of the water quality testing on samples collected from the City’s wastewater treatment plants. 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 increase PHB yield. Marko also encouraged us to consider the upfront costs of incorporating our system into existing wastewater facilities, and whether using pure cultures was the best decision. He mentioned that the wastewater treatment industry already uses methods for production of value-added products like methane and fertilizers and that those processes do not require as much equipment or as many steps when compared to our the PHB production process. This made it less likely that our project would successfully be implemented in wastewater treatment plants, which encouraged us to look into our other possible applications.

DR. DARINA KUZMA

Profession: ACWA Analytical Lab Manager
ACWA
ACWA (Advancing Canadian Wastewater Assets) analyzes water that has already gone through the sanitation process, and their research focuses on finding ways of eliminating particles that are too small for regular methods to detect (such as over-the-counter medication and antibiotics). Dr. Kuzma mentioned that if our secreted PHB particles were too small, we would need to carefully consider the environmental safety of our process if these particles left the containment of a wastewater treatment plant and 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 enters municipal 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 to operate. If we applied our PHB production to landfills, they would have to economically compete with deep well injections. Due to the current low demand for PHB plastics, this feat might not have been easily accomplished. 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 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 cellular 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 via gene knockout or providing our bacteria with alternate nutrient media.

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 our plans for developing a mathematical 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. This helped us to work toward a model that could closely simulate the lab experiments. He also referred us to Dr. Lewis, whose lab studies metabolomics and flux balance analysis in model organisms.

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 our 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(s) that E. coli will use after inserting our genetic constructs. Furthermore, the team was told about how flux balance analysis could progress into flux variability analysis, which would help us to modify parameters in the model and analyze changes in PHB production under a variety of conditions.

DR. JASON DE KONING

Profession: Assistant Professor, Bioinformatics, Biochemistry and Molecular Biology
University of Calgary
In our meeting with Dr. De Koning, he told us about codon optimization and how it could be used to improve expression of our constructs. Thus, the team decided to codon optimize all of our BioBricks submitted to the Registry. We also discussed potential models for the project with Dr. De Koning, and e provided us with resources and software to help shape the parameters of our kinetic model.

DR. NASHAAT NASSAR

Profession: Associate Professor, Chemical and Petroleum Engineering
University of Calgary
Dr. Nassar provided an expert opinion on scaling up our PHB extraction methods. His experience with nanoparticles and adaptation of polymer extraction methods to larger scale helped us to critically evaluate our design choices and come up with our final solution. More details about how we evaluated our design choices based on his feedback can be found on the Extraction page. His insight was also invaluable in designing our coagulation experiments and evaluating how they could be applied on Mars. Our coagulation experiments are described here.

DR. SAURABH JYOTI SARMA

Profession: Postdoctoral Scholar
University of Calgary
As we were considering different applications for our PHB production system in the preliminary stages of the project, Dr. Saurabh provided key insights on employing our project at wastewater treatment plants. He has extensive experience working with volatile fatty acids present in wastewater, and he advised us on the feasibility of using wastewater as feedstock for our genetically engineered bacteria. He raised concerns about the economic viability of adapting our process in wastewater treatment plants, as multimillion dollar changes would have to be made to the general infrastructure and would need to be recovered through municipal taxation and profit of PHB sales. His advice helped us evaluate the applications for our project. More details about how we assessed the various applications can be found on the Human Practices Silver page.