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All sourcesNASA NTRSDOE OSTI
NASA NTRS2017

Methane Metabolism by Yeast for Solar System Exploration

Exploration of the solar system is constrained by the cost of moving mass off Earth. Producing materials in situ will reduce the mass that must be delivered from earth. CO2 is abundant on Mars and manned spacecraft. On the ISS, NASA reacts excess CO2 with H2 to generate CH4 and H2O using the Sabatier System. The resulting water is recovered into the ISS, but the methane is vented to space. Thus, there is a capability need for systems that convert methane into valuable materials. Methanotrophic bacteria consume methane but these are poor synthetic biology platforms. Thus, there is a knowledge gap in utilizing methane in a robust and flexible synthetic biology platform. The yeast Pichia pastoris is a refined microbial factory that is used widely by industry because it efficiently secretes products. Pichia could produce a variety of useful products in space. Pichia does not consume methane but robustly consumes methanol, which is one enzymatic step removed from methane. Our goal is to engineer Pichia to consume methane thereby creating a powerful methane-consuming microbial factory.

Galazka, Jonathan M.↗
NASA NTRSDate not supplied

New Frontiers in Synthetic Biology for Spaceflight

Exploration of the solar system is constrained by the cost of moving mass off Earth. Producing materials in situ will reduce the mass that must be delivered from earth. CO2 is abundant on Mars and manned spacecraft. On the ISS, NASA reacts excess CO2 with H2 to generate CH4 and H2O using the Sabatier System. The resulting water is recovered into the ISS, but the methane is vented to space. Thus, there is a capability need for systems that convert methane into valuable materials. Methanotrophic bacteria consume methane but these are poor synthetic biology platforms. Thus, there is a knowledge gap in utilizing methane in a robust and flexible synthetic biology platform. The yeast Pichia pastoris is a refined microbial factory that is used widely by industry because it efficiently secretes products. Pichia could produce a variety of useful products in space. Pichia does not consume methane but robustly consumes methanol, which is one enzymatic step removed from methane. Our goal is to engineer Pichia to consume methane thereby creating a powerful methane-consuming microbial factory.

methane↗
NASA NTRSDate not supplied

Final Report of the COSPAR Meeting Series on Knowledge Gaps in Planetary Protection for Crewed Missions to Mars

The international scientific community has, through a Committee on Space Research (COSPAR) meeting series, participated in a seven-year review of knowledge gaps in planetary protection for crewed missions to Mars. This meeting series first identified a prioritized list of knowledge gaps that, if addressed, would permit development of an implementable planetary protection policy for the initial crewed Mars mission. Next, a consideration of the available venues (ground, LEO, deep space, planetary surfaces) yielded a roadmap of potential future investigations needed to close the knowledge gaps. Finally, the meeting series evaluated topic-specific measurements that are essential to establishing the figures of merit needed for hardware engineering design requirements. The three main topic threads considered within the meeting series were: microbial and human health monitoring; natural transport of terrestrial biological contamination on Mars, and; technologies and operations for contamination control (of spacecraft systems). Participants at the meeting series included engineers, scientists, managers and various subject matter experts from space agencies, academic and research institutions, commercial companies, and other government departments. This paper provides a detailed summary review of the findings of the meeting series, including descriptions of the basis for each of the figures of merit (where they were identified), and considerations of how planetary protection implementation and policy for crewed missions may progress from the current state of the art.

J Andy Spry↗
NASA NTRS1981

Evaluation of engineered foods for Closed Ecological Life Support System (CELSS)

A system of conversion of locally regenerated raw materials and of resupplied freeze-dried foods and ingredients into acceptable, safe and nutritious engineered foods is proposed. The first phase of the proposed research has the following objectives: (1) evaluation of feasibility of developing acceptable and reliable engineered foods from a limited selection of plants, supplemented by microbially produced nutrients and a minimum of dehydrated nutrient sources (especially those of animal origin); (2) evaluation of research tasks and specifications of research projects to adapt present technology and food science to expected space conditions (in particular, problems arising from unusual gravity conditions, problems of limited size and the isolation of the food production system, and the opportunities of space conditions are considered); (3) development of scenarios of agricultural production of plant and microbial systems, including the specifications of processing wastes to be recycled.

Karel, M.↗
NASA NTRSDate not supplied

Spaceflight Autonomous Multigenerational Microbial Sequencer in Support of Plant-Growth Systems

The CubeSat platform has proven successful in obtaining meaningful life science information when biological payloads are incorporated. Examples include: 1) the first-ever CubeSat with a biological payload, GeneSat-1, which demonstrated decreased growth rates for flight samples of Escherichia coli in low Earth orbit (Parra et al. 2008); 2) PharmaSat, demonstrated that Saccharomyces cerevisiae in the microgravity environment exhibits a significant level of metabolic activity even at high doses of applied antifungal (Ricco et al. 2011); 3) O/OREOS, which used Bacillus subtilis(bacteria) to demonstrate for the first time that microorganisms can be loaded in a dried, dormant form and then rehydrated and grown in orbit months after launch (Nicholson et al. 2011; Ehrenfreund et al. 2014; 4) the SporeSat payload, which investigated Ceratopteris richardii(fern spores) using lab-on-a-chip devices (BioCDs) and minicentrifuges to produce artificial gravitational forces in ground studies (Park et al. 2017), with demonstration of the BioCD and minicentrifuge in space; 5) EcAMSat, the first CubeSat to be directly deployed from the ISS for an experiment assessing antibiotic resistance of E. coli in the microgravity environment (Padgen et al. 2020); 6) BioSentinel, exposed a culture of yeast to galactic cosmic radiation (GCR) and solar particle events while in heliocentric orbit to measure the rate of double-strand-break repair using DNA-repair-deficient mutants. This effort measures the metabolic parameters of yeast in a deep-space environment compared to Earth ambient conditions using a 3-color LED detection system (Ricco et al. 2020; Padgen et al. 2021). We aim to expand this list to include a Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS). SAMMS will allow for the genome level understanding of changes in growth and metabolic activity for any organism. While microbes are suitable for early studies in our proposed platform because of their small size, small and relatively less-complicated genomes, fast generation times, and relevance to life support systems; multicellular organisms can similarly be evaluated for their genetic response to the spaceflight environment. The Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) will enable autonomous sequencing of biological samples in plant production units, cislunar orbit and on the lunar surface to examine spaceflight effects (ie. radiation, altered gravity, reduced pressures) on plant and microbial genomes.On this team a Kennedy Space Center (KSC) space crop production and water systems microbiologist/molecular biologist works with a Johnson Space Center (JSC) International Space Station (ISS) microbial sequencing expert and an Ames Research Center (ARC) CubeSat Engineering team to convert an automated Oxford Nanopore librarypreparation and sequencing method to a fluidic CubeSat payload system. The Oxford Nanopore MinION sequencing platform has proven successful in the spaceflight environment onboard the ISS (Stahl-Rommel et al. 2021). Further long-duration spaceflight and exposure to high levels of radiation will cause genotypic effects in biological organisms that may affect their function. Monitoring the adaption of a population to the spaceflight environment and any subsequent beneficial mutations will allow for the harnessing of organisms best suited for use in life support systems. This will ensure that the selected life support-essential microorganisms maintain their intended specified function over generations of culturing in the relevant spaceflight environment without becoming hazardous to crew or spacecraft systems.

Aubrie E Orourke↗
NASA NTRSDate not supplied

Planetary Protection, Parts, and Perception: The Ranger Missions as the Origin Story of the Myth of Planetary Protection-Related Mission Failures

There has been a long-standing perspective in the planetary engineering community that the use of planetary protection sterilization techniques such as dry heat microbial reduction (DHMR) have led to significant mission failures in the past. This perception has led to a hesitation to craft mission designs that incorporate sterilization beyond the component-level. Via interviews with long-standing members of the planetary engineering community across the Agency, we have identified the root historical missions that have driven this perspective. The source, or Origin Story, which has been passed down to subsequent generations of engineers, is the failures tied to the early series of the Ranger missions. In this presentation, we examine the details behind the Ranger failures, the majority of which are non-planetary protection sterilization-related, and bring to light the myth of this perception.

planetary protection↗
NASA NTRS2015

Human Factors and Habitability Challenges for Mars Missions

As NASA is planning to send humans deeper into space than ever before, adequate crew health and performance will be critical for mission success. Within the NASA Human Research Program (HRP), the Space Human Factors and Habitability (SHFH) team is responsible for characterizing the risks associated with human capabilities and limitations with respect to long-duration spaceflight, and for providing mitigations (e.g., guidelines, technologies, and tools) to promote safe, reliable and productive missions. SHFH research includes three domains: Advanced Environmental Health (AEH), Advanced Food Technology (AFT), and Space Human Factors Engineering (SHFE). The AEH portfolio focuses on understanding the risk of microbial contamination of the spacecraft and on the development of standards for exposure to potential toxins such as chemicals, bacteria, fungus, and lunar/Martian dust. The two risks that the environmental health project focuses on are adverse health effects due to changes in host-microbe interactions, and risks associated with exposure to dust in planetary surface habitats. This portfolio also proposes countermeasures to these risks by making recommendations that relate to requirements for environmental quality, foods, and crew health on spacecraft and space missions. The AFT portfolio focuses on reducing the mass, volume, and waste of the entire integrated food system to be used in exploration missions, and investigating processing methods to extend the shelf life of food items up to five years, while assuring that exploration crews will have nutritious and palatable foods. The portfolio also delivers improvements in both the food itself and the technologies for storing and preparing it. SHFE sponsors research to establish human factors and habitability standards and guidelines in five risk areas, and provides improved design concepts for advanced crew interfaces and habitability systems. These risk areas include: Incompatible vehicle/habitat design, inadequate human-computer interaction, inadequate critical task design, inadequate human-automation/robotic interaction, and performance errors due to training deficiencies. To address the identified research gaps within each risk, SHFH's research plan includes studies in the laboratory, in analogs, and on International Space Station (ISS). In addition to establishing and maintaining the risk-based research portfolio, SHFH is also implementing a qualitative approach to determine how we at NASA evaluate human performance. Via interviews with experts, such as trainers, flight controllers, and flight surgeons, we are collecting the metrics by which they assess human performance, evidence of performance issues, and potential or actual consequences. The Human Performance Data Project will determine what human performance data have been collected in the past at NASA, and what data should be collected in the future in order to complete our knowledgebase and reduce risks related to human factors and habitability.

Whitmore, Mihriban↗
NASA NTRSDate not supplied

Ground Test Validations for Uncrewed Dormancy Preparation and Recovery

In future long duration space flight and habitation missions, periods of uncrewed dormancy where life support systems are shutdown or in low power-modes will occur. To examine the impacts of these dormant periods, along with potential pre- and post-dormancy procedures, Marshall Space Flight Center has been building an uncrewed dormancy test stand. The uncrewed dormancy test stand will include a urine separator from the Universal Waste Management System, a condensate separator, urine filter, brine filter and distillate filter. In conjunction to the test stand, the Center for Biofilm Engineering (CBE) has ongoing long-term dormancy experiments to assess microbial survival in a variety of simulated shutdown procedures including shut-in, flushing with and without biocide, and dry-out conditions. This publication will serve as a discussion of dormancy considerations, criteria for success, and experimental designs for testing dormancy conditions and shutdown procedures. It will include an update of ongoing test stand construction, including a description of the current lower-fidelity versions of the urine and condensate separators being produced. It will also examine dormancy work being conducted by the CBE and discuss how this may factor into procedures tested on the uncrewed dormancy test stand.

Eric Beitle↗
NASA NTRS2018

EVA Swab Tool to Support Planetary Protection and Astrobiology Evaluations

When we send humans to search for life on other planets, we'll need to know what we brought with us versus what may already be there. To ensure our crewed systems meet planetary protection requirements-and to protect our science from human contamination-we'll need to assess whether microorganisms may be leaking or venting from our spacecraft. Microbial sample collection outside of a pressurized spacecraft is complicated by temperature extremes, low pressures that preclude the use of laboratory standard (wetted) swabs, and operation either in bulky spacesuits or with robotic assistance. Engineers at the National Aeronautics and Space Administration (NASA) recently developed a swab kit for use in collecting microbial samples from the external surfaces of crewed spacecraft, including spacesuits. The Extravehicular Activity (EVA) Swab Kit consists of a single swab tool handle and an eight-canister sample caddy. The design team minimized development cost by re-purposing a heritage Space Shuttle tile repair handle that was designed to quickly snap into different tool attachments by engaging a mating device in each attachment. This allowed the tool handle to snap onto a fresh swab attachment much like popular shaving razor handles can snap onto a disposable blade cartridge. To disengage the handle from a swab, the user performs two independent functions, which can be done with a single hand. This dual operation mitigates the risk that a swab will be inadvertently released and lost in microgravity. Each swab attachment is fitted with commercially available foam swab tips, vendor-certified to be sterile for Deoxyribonucleic Acid (DNA). A microbial filter installed in the bottom of each sample container allows the container to outgas and repressurize without introducing microbial contaminants to internal void spaces. Extensive ground testing, post-test handling, and sample analysis confirmed the design is able to maintain sterile conditions as the canister moves between various pressure environments. To further minimize cost, the design team acquired extensive ground test experience in a relevant flight environment by piggy-backing onto suited crew training runs. These training runs allowed the project to validate tool interfaces with pressurized EVA gloves and collect user feedback on the tool design and function, as well as characterize baseline microbial data for different types of spacesuits. In general, test subjects found the EVA Swab Kit relatively straightforward to operate, but identified a number of design improvements that will be incorporated into the final design. Although originally intended to help characterize human forward contaminants, this tool has other potential applications, such as for collecting and preserving space-exposed materials to support astrobiology experiments.

Rucker, Michelle A.↗
NASA NTRS2013

NASA Tech Briefs: What Does a Microbial Ecologist Do?

Dr. Leslie Bebout works as a microbial ecologist in the Exobiology Branch at NASA's Moffett Field, CA-­‐based Ames Research Center. She and her colleagues study the complexities of carbon, nitrogen and hydrogen cycling in early Earth and Mars analog microbial systems. They concurrently are using this systems biology approach to work with engineers to design systems geared to optimize the use of water, light and nutrient resources relevant both to the development of new green technologies and space exploration capabilities.

Garud, Sumedha↗
NASA NTRS2013

Microbial Monitoring of the International Space Station

Humans living and working in the harsh environment of space present many challenges for habitability engineers and microbiologists. Spacecraft must provide an internal environment in which physical (gas composition, pressure, temperature, and humidity), chemical, and biological environmental parameters are maintained at safe levels. Microorganisms are ubiquitous and will accompany all human-occupied spacecraft, but if biological contamination were to reach unacceptable levels, long-term human space flight would be impossible. Prevention of microbiological problems, therefore, must have a high priority. Historically, prevention of infectious disease in the crew has been the highest priority, but experience gained from the NASA-Mir program showed that microbial contamination of vehicle and life-support systems, such as biofouling of water and food, are of equal importance. The major sources of microbiological risk factors for astronauts include food, drinking water, air, surfaces, payloads, research animals, crew members, and personnel in close contact with the astronauts. In our efforts to eliminate or mitigate the negative effects of microorganisms in spacecraft, the National Aeronautics and Space Administration (NASA) implemented comprehensive microbial analyses of the major risk factors. This included the establishment of acceptability requirements for food, water, air, surfaces, and crew members. A robust monitoring program was then implemented to verify that the risks were within acceptable limits. Prevention of microbiological problems is preferred over mitigation of problems during flight, and preventive steps must begin very early in the design phase. Spacecraft development must include requirements to control free water from humidity, condensate, hygiene activities, and other releases. If water is available, microbes are likely to grow because sufficient nutrients are potentially available. Materials selected for the spacecraft must not promote or support microbial growth. Air filtration can dramatically reduce the number of airborne bacteria, fungi, and particulates in spacecraft breathing air. Waterborne bacteria can be reduced to acceptable levels by thermal inactivation of bacteria during water processing, along with a residual biocide, and filtration at the point of use can ensure safety. System design must include onboard capability to achieve recovery of the system from contamination. Robust housekeeping procedures that include periodic cleaning and disinfection will prevent high levels of microbial growth on surfaces. Food for consumption in space must be thoroughly tested for excessive microbial content and pathogens before launch. Thorough preflight examination of flight crews, consumables, payloads, and the environment can greatly reduce pathogens in spacecraft. Many of the lessons learned from the Space Shuttle and previous programs were applied in the early design phase of the International Space Station, resulting in the safest space habitat to date. This presentation describes the monitoring program for the International Space Station and will summarize results from preflight and on-orbit monitoring.

Pierson, Duane L.↗
NASA NTRS2006

Planned Environmental Microbiology Aspects of Future Lunar and Mars Missions

With the establishment of the Constellation Program, NASA has initiated efforts designed similar to the Apollo Program to return to the moon and subsequently travel to Mars. Early lunar sorties will take 4 crewmembers to the moon for 4 to 7 days. Later missions will increase in duration up to 6 months as a lunar habitat is constructed. These missions and vehicle designs are the forerunners of further missions destined for human exploration of Mars. Throughout the planning and design process, lessons learned from the International Space Station (ISS) and past programs will be implemented toward future exploration goals. The standards and requirements for these missions will vary depending on life support systems, mission duration, crew activities, and payloads. From a microbiological perspective, preventative measures will remain the primary techniques to mitigate microbial risk. Thus, most of the effort will focus on stringent preflight monitoring requirements and engineering controls designed into the vehicle, such as HEPA air filters. Due to volume constraints in the CEV, in-flight monitoring will be limited for short-duration missions to the measurement of biocide concentration for water potability. Once long-duration habitation begins on the lunar surface, a more extensive environmental monitoring plan will be initiated. However, limited in-flight volume constraints and the inability to return samples to Earth will increase the need for crew capabilities in determining the nature of contamination problems and method of remediation. In addition, limited shelf life of current monitoring hardware consumables and limited capabilities to dispose of biohazardous trash will drive flight hardware toward non-culture based methodologies, such as hardware that rapidly distinguishes biotic versus abiotic surface contamination. As missions progress to Mars, environmental systems will depend heavily on regeneration of air and water and biological waste remediation and regeneration systems, increasing the need for environmental monitoring. Almost complete crew autonomy will be needed for assessment and remediation of contamination problems. Cabin capacity will be limited; thus, current methods of microbial monitoring will be inadequate. Future methodology must limit consumables, and these consumables must have a shelf life of over three years. In summary, missions to the moon and Mars will require a practical design that prudently uses available resources to mitigate microbial risk to the crew.

Ott, C. Mark↗
NASA NTRS2017

Impact of Biofilms on the Design and Operation of ISS Life Support Systems

Biofilm growth has been an ongoing issue for US and Russian water systems on the International Space Station, and is a critical issue for exploration missions in which water systems must be designed to accommodate dormant periods of up to one year. On ISS, Russian condensate plumbing has previously clogged with biomass, requiring condensate plumbing to now be regularly replaced. In the US Segment, the release of biofilm from the Water Processor waste tank has clogged a solenoid valve downstream of the tank, resulting in the costly replacement of the inlet separator and process pump. Subsequent management of the biofilm in the waste tank involves restrictions on tank cycles to limit the release of biomass and an additional filter to protect downstream components. Engineering personnel are now evaluating concepts to better manage the biomass, including the use of microbial inhibitors and UV LEDs. Though current ISS operations could likely be sustained for the duration of ISS, a more effective method must be developed for managing the growth and release of biomass in future exploration vehicles. Biofilm management for future missions is complicated by the requirement to accommodate extended periods of dormancy during which time the water system will be stagnant. The current approach under consideration is to flush the waste water with product water to reduce the organic content followed by use of microbial inhibitors or UV. However, other concepts may also be developed based on ongoing research.

Carter, Donald Layne↗
NASA NTRS2006

The Validation of Vapor Phase Hydrogen Peroxide Microbial Reduction for Planetary Protection and a Proposed Vacuum Process Specification

The Jet Propulsion Laboratory, in conjunction with the NASA Planetary Protection Officer, has selected the vapor phase hydrogen peroxide sterilization process for continued development as a NASA approved sterilization technique for spacecraft subsystems and systems. The goal is to include this technique, with an appropriate specification, in NPR 8020.12C as a low temperature complementary technique to the dry heat sterilization process.To meet microbial reduction requirements for all Mars in-situ life detection and sample return missions, various planetary spacecraft subsystems will have to be exposed to a qualified sterilization process. This process could be the elevated temperature dry heat sterilization process (~115 C for 40 hours) which was used to sterilize the Viking lander spacecraft. However, with utilization of such elements as highly sophisticated electronics and sensors in modern spacecraft, this process presents significant materials challenges and is thus an undesirable bioburden reduction method to design engineers. The objective of this work is to introduce vapor hydrogen peroxide (VHP) as an alternative to dry heat microbial reduction to meet planetary protection requirements.The VHP process is widely used by the medical industry to sterilize surgical instruments and biomedical devices, but high doses of VHP may degrade the performance of flight hardware, or compromise material properties. Our goal for this study was to determine the minimum VHP process conditions to achieve microbial reduction levels acceptable for planetary protection.

planetary protection↗
NASA NTRS2013

An Assessment of Environmental Health Needs for Manned Spacecraft

Environmental health fundamentally addresses the physical, chemical, and biological risks external to the human body that can impact the health of a person by assessing and controlling these risks in order to generate and maintain a health‐supportive environment. Environmental monitoring coupled with other measures including active and passive controls and the implementation of environmental standards (SMACs, SWEGs, microbial and acoustics limits) are used to ensure environmental health in manned spacecraft. NASA scientists and engineers consider environmental monitoring a vital component to an environmental health management strategy for maintaining a healthy crew and achieving mission success. Environmental monitoring data confirms the health of ECLS systems, in addition to contributing to the management of the health of human systems. Crew health risks associated with the environment were reviewed by agency experts with the goal of determining risk-based environmental monitoring needs for future NASA manned missions. Once determined, gaps in knowledge and technology, required to address those risks, were identified for various types of Exploration missions. This agency‐wide assessment of environmental health needs will help guide the activities/hardware development efforts to close those gaps and advance the knowledge required to meet NASA manned space exploration objectives. Details of this assessment and findings are presented in this paper.

Macatangay, Ariel V.↗
NASA NTRS2000

Morphology and Viability of Pleistocene Microbiota from the CRREL Permafrost Tunnel Near Fox, Alaska

The U. S. Army Cold Regions Research and Engineering Laboratory maintains the CRREL Permafrost Tunnel at Fox, Alaska (-10 miles north of Fairbanks.) The active microbial ecosystems and the cryopreserved anabiotic viable microorganisms and dead microbial remains and biomarkers frozen within the permafrost and ice of the CRREL Permafrost Tunnel are of direct relevance to Astrobiology. Microbial extremophiles from permafrost and ice provide information concerning where and how should we search for evidence of life elsewhere in the Cosmos. The permafrost and ice wedges of the Fox tunnel preserves a magnificent of record of Pliocene, Pleistocene and Holocene life on Earth spanning more than 2.5 million years. This record includes frozen fossil bacteria, archaea, algae, mosses, higher plants, insects and mammals. In this paper we present the preliminary results of studies of the morphology, ultramicrostructure and elemental distributions of Fox tunnel microbiota as determined in-situ by the Environmental Scanning Electron Microscope (ESEM) and the Field Emission Scanning Electron Microscope (FESEM) investigations. The long-term viability of cryopreserved microbiota and potential implications to Astrobiology will be discussed.

Hoover, Richard B.↗
NASA NTRS2011

Post-Fragmentation Whole Genome Amplification-Based Method

This innovation is derived from a proprietary amplification scheme that is based upon random fragmentation of the genome into a series of short, overlapping templates. The resulting shorter DNA strands (<400 bp) constitute a library of DNA fragments with defined 3 and 5 termini. Specific primers to these termini are then used to isothermally amplify this library into potentially unlimited quantities that can be used immediately for multiple downstream applications including gel eletrophoresis, quantitative polymerase chain reaction (QPCR), comparative genomic hybridization microarray, SNP analysis, and sequencing. The standard reaction can be performed with minimal hands-on time, and can produce amplified DNA in as little as three hours. Post-fragmentation whole genome amplification-based technology provides a robust and accurate method of amplifying femtogram levels of starting material into microgram yields with no detectable allele bias. The amplified DNA also facilitates the preservation of samples (spacecraft samples) by amplifying scarce amounts of template DNA into microgram concentrations in just a few hours. Based on further optimization of this technology, this could be a feasible technology to use in sample preservation for potential future sample return missions. The research and technology development described here can be pivotal in dealing with backward/forward biological contamination from planetary missions. Such efforts rely heavily on an increasing understanding of the burden and diversity of microorganisms present on spacecraft surfaces throughout assembly and testing. The development and implementation of these technologies could significantly improve the comprehensiveness and resolving power of spacecraft-associated microbial population censuses, and are important to the continued evolution and advancement of planetary protection capabilities. Current molecular procedures for assaying spacecraft-associated microbial burden and diversity have inherent sample loss issues at practically every step, particularly nucleic acid extraction. In engineering a molecular means of amplifying nucleic acids directly from single cells in their native state within the sample matrix, this innovation has circumvented entirely the need for DNA extraction regimes in the sample processing scheme.

Benardini, James↗
NASA NTRS2017

Engineering of Methane Metabolism in Pichia Pastoris Through Methane Monooxygenase Expression

Exploration of the solar system is constrained by the cost of moving mass off Earth. Producing materials in situ will reduce the mass that must be delivered from earth. CO2 is abundant on Mars and manned spacecraft. On the ISS, NASA reacts excess CO2 with H2 to generate CH4 and H2O using the Sabatier System. The resulting water is recovered into the ISS, but the methane is vented to space. Thus, there is a capability need for systems that convert methane into valuable materials. Methanotrophic bacteria consume methane but these are poor synthetic biology platforms. Thus, there is a knowledge gap in utilizing methane in a robust and flexible synthetic biology platform. The yeast Pichia pastoris is a refined microbial factory that is used widely by industry because it efficiently secretes products. Pichia could produce a variety of useful products in space. Pichia does not consume methane but robustly consumes methanol, which is one enzymatic step removed from methane. Our goal is to engineer Pichia to consume methane thereby creating a powerful methane-consuming microbial factory.

Pichia↗