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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 145 records · Page 8
All sourcesNASA NTRSDOE OSTI
DOE OSTI2020

Media Robustness and Scalability of Phosphate Regulated Promoters Useful for Two-Stage Autoinduction in E. coli

A key challenge in synthetic biology is the successful utilization of characterized parts, such as promoters, in different biological contexts. We report the evaluation of the media robustness of a small library of E. coli PhoB regulated promoters that enable heterologous protein production in two-stage cultures. Expression levels were measured both in a rich Autoinduction Broth as well as a minimal mineral salts media. Expression was both media and promoter dependent. Of the 16 promoters tested, 4 were identified to have tightly controlled expression, which was also robust to media formulation. Improved promoter robustness led to more predictable scale up and consistent expression in instrumented bioreactors. Furthermore, this subset of PhoB activated promoters, useful for two-stage autoinduction, highlights the impact of the environment on the performance of biological parts and the importance of robustness testing in synthetic biology.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

The pGinger Family of Expression Plasmids

Metabolic engineering and synthetic biology are predicated on the precise control of gene expression. As synthetic biology expands beyond model organisms, more tools will be required that function robustly in a wide range of bacterial hosts. The pGinger family of plasmids constitutes 43 plasmids that will enable both constitutive and inducible gene expression in a wide range of nonmodel Proteobacteria .

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Translating advances in microbial bioproduction to sustainable biotechnology

Advances in synthetic biology have radically changed our ability to rewire microorganisms and significantly improved the scalable production of a vast array of drop-in biopolymers and biofuels. The success of a drop-in bioproduct is contingent on market competition with petrochemical analogues and weighted upon relative economic and environmental metrics. While the quantification of comparative trade-offs is critical for accurate process-level decision making, the translation of industrial ecology to synthetic biology is often ambiguous and assessment accuracy has proven challenging. In this review, we explore strategies for evaluating industrial biotechnology through life cycle and techno-economic assessment, then contextualize how recent developments in synthetic biology have improved process viability by expanding feedstock availability and the productivity of microbes. By juxtaposing biological and industrial constraints, we highlight major obstacles between the disparate disciplines that hinder accurate process evaluation. The convergence of these disciplines is crucial in shifting towards carbon neutrality and a circular bioeconomy.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2026

Causes and consequences of experimental variation in Nicotiana benthamiana transient expression

Infiltration of Agrobacterium tumefaciens into Nicotiana benthamiana has become a foundational technique in plant biology, enabling efficient delivery of transgenes in planta with technical ease, robust signal, and relatively high throughput. Despite transient expression’s prevalence in disciplines such as synthetic biology, little work has been done to describe and address the variability inherent in this system, a concern for experiments that rely on highly quantitative readouts. In a comprehensive analysis of N. benthamiana agroinfiltration experiments, we model sources of variability that affect transient expression. Our findings emphasize the need to validate normalization methods under the specific conditions of each study, as distinct normalization schemes do not always reduce variation either within or between experiments. Using a dataset of 1915 plants collected over three years, we develop a model of variation in N. benthamiana transient expression, using power analysis to determine the number of individual plants required for a given effect size. Drawing on our longitudinal data, these findings inform practical guidelines for minimizing variability through strategic experimental design and power analysis, providing a foundation for more robust and reproducible use of N. benthamiana in quantitative plant biology and synthetic biology applications.

Tang, Sophia N. [Joint BioEnergy Institute (JBEI),↗
NASA NTRS2020

Life Beyond Earth: Past, Present and Future

The key to life anywhere, including off planet, will be focusing in an ecological approach, both understanding the relationship of the organisms to their environment, and to each other. This includes the fundamental questions of astrobiology: “Where do we come from?”, “Where are we going?” and “Are we alone?” For over a decade NASA has used synthetic biology, which is in simplest form, is creating new capabilities with life, as an enabling technology to answer these questions. Examples will be given for the search for life elsewhere, and notably, the use of synthetic biology as an enabler of human exploration. We have a current satellite mission that combines synthetic biology and the first steps towards enabling a human colony off planet using an ecological approach.

Rothschild, Lynn J.↗
NASA NTRSDate not supplied

Life Beyond Earth: Past, Present and Future

The key to life anywhere, including off planet, will be focusing in an ecological approach, both understanding the relationship of the organisms to their environment, and to each other. This includes the fundamental questions of astrobiology: “Where do we come from?”, “Where are we going?” and “Are we alone?” For over a decade NASA has used synthetic biology, which is in simplest form, is creating new capabilities with life, as an enabling technology to answer these questions. Examples will be given for the search for life elsewhere, and notably, the use of synthetic biology as an enabler of human exploration. We have a current satellite mission that combines synthetic biology and the first steps towards enabling a human colony off planet using an ecological approach.

Lynn J Rothschild↗
DOE OSTI2021

Unnatural biosynthesis by an engineered microorganism with heterologously expressed natural enzymes and an artificial metalloenzyme

Synthetic biology enables microbial hosts to produce complex molecules from organisms that are rare or difficult to cultivate, but the structures of these molecules are limited to those formed by reactions of natural enzymes. The integration of artificial metalloenzymes (ArMs) that catalyse unnatural reactions into metabolic networks could broaden the cache of molecules produced biosynthetically. Here, in this work, we report an engineered microbial cell expressing a heterologous biosynthetic pathway, containing both natural enzymes and ArMs, that produces an unnatural product with high diastereoselectivity. We engineered Escherichia coli with a heterologous terpene biosynthetic pathway and an ArM containing an iridium–porphyrin complex that was transported into the cell with a heterologous transport system. We improved the diastereoselectivity and product titre of the unnatural product by evolving the ArM and selecting the appropriate gene induction and cultivation conditions. This work shows that synthetic biology and synthetic chemistry can produce, by combining natural and artificial enzymes in whole cells, molecules that were previously inaccessible to nature.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

Development of a Freeze-Dried CRISPR-Cas12 Sensor for Detecting Wolbachia in the Secondary Science Classroom

Training the future synthetic biology workforce requires the opportunity for students to be exposed to biotechnology concepts and activities in secondary education. Detecting Wolbachia bacteria in arthropods using polymerase chain reaction (PCR) has become a common way for secondary students to investigate and apply recombinant DNA technology in the science classroom. Despite this important activity, cutting-edge biotechnologies such as clustered regularly interspaced short palindromic repeat (CRISPR)-based diagnostics have yet to be widely implemented in the classroom. To address this gap, we present a freeze-dried CRISPR-Cas12 sensing reaction to complement traditional recombinant DNA technology education and teach synthetic biology concepts. The reactions accurately detect Wolbachia from arthropod-derived PCR samples in under 2 h and can be stored at room temperature for over a month without appreciable degradation. Here, the reactions are easy-to-use and cost less than $40 to implement for a classroom of 22 students including the cost of reusable equipment. We see these freeze-dried CRISPR-Cas12 reactions as an accessible way to incorporate synthetic biology education into the existing biology curriculum, which will expand biology educational opportunities in science, technology, engineering, and mathematics.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Optimization of C. ljungdahlii for the Valorization of Combustion Off-Gas

One of the largest problems that faces the realization of the circular economy is the gap between waste streams and usable commodities. Synthetic biology enables a toolkit where this gap can begin to be addressed and closed. This work focuses on using synthetic biology techniques to optimize C. ljungdahlii for the valorization of combustion off-gas waste streams and navigates how to produce 3-hydroxybutyrate, a polymer precursor off of similar-composition syn-gas streams. If successful, these techniques can realize a carbon-negative solution to producing plastics.

3-hydroxybutyrate↗
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↗
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.

Methane↗
NASA NTRS2015

Enhanced Polyhydroxybutyrate Production for Long-Term Spaceflight Applications

Synthetic biology holds the promise of advancing long term space fight by the production of medicine, food, materials, and energy. One such application of synthetic biology is the production of biomaterials, specifically polyhydroxyalkanoates (PHAs), using purposed organisms such as Escherichia coli. PHAs are a group of biodegradable bioplastics that are produced by a wide variety of naturally occurring microorganisms, mainly as an energy storage intermediate. PHAs have similar melting point to polypropylene and a Youngs modulus close to polystyrene. Due to limited resources and cost of transportation, large-scale extraction of biologically produced products in situ is extremely cumbersome during space flight. To that end, we are developing a secretion systems for exporting PHA from the cell in order to reduce unit operations. PHAs granules deposited inside bacteria are typically associated with proteins bound to the granule surface. Phasin, a granule bound protein, was targeted for type I secretion by fusion with HlyA signal peptide for indirect secretion of PHAs. In order to validate our secretion strategy, a green fluorescent protein (GFP) was tagged to the PHA polymerase enzyme (phaC), this three part gene cassette consists of phaA and phaB and are required for PHA production. Producing PHAs in situ during space flight or planet colonization will enable mission success by providing a valuable source of biomaterials that can have many potential applications thereby reducing resupply requirements. Biologically produced PHAs can be used in additive manufacturing such as three dimensional (3D) printing to create products that can be made on demand during space flight. After exceeding their lifetime, the PHAs could be melted and recycled back to 3D print other products. We will discuss some of our long term goals of this approach.

In Situ↗
NASA NTRS2013

Evolution to Space

This presentation will discuss recent space exploration results (LCROSS, KEPLER, etc.), increase access to space and the small and cube satellites platform as it relates to the future of space exploration. It will highlight the concept of modularization and the use of biology, and specifically synthetic biology in the future. The presentation will be a general public presentation. When speaking to a younger audience, I will discuss my background. All slides contain only public information. No technical ITAR/Export controlled material will be discussed.

Cohen, Jacob↗
NASA NTRS2019

Results of the Micro-12 Flight Experiment: Effects of Microgravity on Shewanella oneidensis MR-1

The Micro-12 flight experiment was launched on SpaceX-15 and completed during berthing on the International Space Station. The goal of this experiment was to understand the effects of spaceflight and microgravity on the physiology of the model exoelectrogen Shewanella oneidensis MR-1. BioServe Fluid Processing Apparatus (FPA) and Group Activation Pack (GAP) hardware systems were used for both flight and ground control tests. Under spaceflight conditions, extracellular electron transfer (EET) rates were found to be significantly increased on insoluble substrates, while biofilm development appeared to be unchanged under the conditions tested; these processes are critical for microbial-assisted bioelectrochemical systems. Additionally, RNAseq analysis, proteomic profiling, and competitive mutant fitness profiling were performed to gain further understanding of microbial physiology under EET-respiring conditions during spaceflight. Overall, the results of the Micro-12 project support the idea that Shewanella oneidensis MR-1, in particular, and exoelectrogens in general could be useful chassis organisms for synthetic biology applications using microbial bioelectrochemical systems. These findings will assist bioengineering and synthetic biology development efforts harnessing the unique capabilities of exoelectrogens for life support and in situ resource utilization.

Dougherty, Michael↗
NASA NTRS2015

“PowerCell”: The Interface Between Mars Resources and Human Exploration

The barriers to forming human settlements on Mars are high but surmountable within our lifetime. While the Apollo astronauts carried their life support with them, our success in exploring and forming settlements on Mars depends on our ability to use local Martian resources to generate the materials and conditions humans need to survive, so-called in situ resource utilization (ISRU). On Earth, biology provides us with food, shelter, oxygen, and other materials. Off-planet, synthetic biology will enable numerous parallel productions: optimized food production, water treatment, air treatment, environmental monitoring, regolith biomining, waste management, cell based biomaterial production, biocementation, and in situ synthesis based on received DNA sequences. How will the organisms responsible for these synthetic production systems obtain organic carbon and fixed nitrogen in the hostile Martian environment? We envision a synthetic-biology enabled Martian colony and introduce here the critical intermediate component a biological power source needed to transform the in situ resources found on Mars into biological feedstocks to enable growth of production organisms. Here, we present our first PowerCell, a photosynthetic and nitrogen-fixing filamentous cyanobacterium engineered to provide a carbon-rich fuel source for a biological life support system on Mars. We provide a vision of how the PowerCell system will operate in a Martian colony based on ground experiments and preparations for testing in space as a NASA secondary payload aboard the upcoming DLR Eu:CROPIS satellite mission experiments.

Rothschild, Lynn J.↗
DOE OSTI2021

Omics-Driven Biotechnology for Industrial Applications

Biomanufacturing is a key component of biotechnology that uses biological systems to produce bioproducts of commercial relevance, which are of great interest to the energy, material, pharmaceutical, food, and agriculture industries. Biotechnology-based approaches, such as synthetic biology and metabolic engineering are heavily reliant on “omics” driven systems biology to characterize and understand metabolic networks. Knowledge gained from systems biology experiments aid the development of synthetic biology tools and the advancement of metabolic engineering studies toward establishing robust industrial biomanufacturing platforms. In this review, we discuss recent advances in “omics” technologies, compare the pros and cons of the different “omics” technologies, and discuss the necessary requirements for carrying out multi-omics experiments. We highlight the influence of “omics” technologies on the production of biofuels and bioproducts by metabolic engineering. Finally, we discuss the application of “omics” technologies to agricultural and food biotechnology, and review the impact of “omics” on current COVID-19 research.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Expanding the genetic engineering toolbox for the metabolically flexible acetogen Eubacterium limosum

Abstract Acetogenic bacteria are an increasingly popular choice for producing fuels and chemicals from single carbon (C1) substrates. Eubacterium limosum is a promising acetogen with several native advantages, including the ability to catabolize a wide repertoire of C1 feedstocks and the ability to grow well on agar plates. However, despite its promise as a strain for synthetic biology and metabolic engineering, there are insufficient engineering tools and molecular biology knowledge to leverage its native strengths for these applications. To capitalize on the natural advantages of this organism, here we extended its limited engineering toolbox. We evaluated the copy number of three common plasmid origins of replication and devised a method of controlling copy number and heterologous gene expression level by modulating antibiotic concentration. We further quantitatively assessed the strength and regulatory tightness of a panel of promoters, developing a series of well-characterized vectors for gene expression at varying levels. In addition, we developed a black/white colorimetric genetic reporter assay and leveraged the high oxygen tolerance of E. limosum to develop a simple and rapid transformation protocol that enables benchtop transformation. Finally, we developed two new antibiotic selection markers—doubling the number available for this organism. These developments will enable enhanced metabolic engineering and synthetic biology work with E. limosum.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

A roadmap to understanding and anticipating microbial gene transfer in soil communities

Engineered microbes are being programmed using synthetic DNA for applications in soil to overcome global challenges related to climate change, energy, food security, and pollution. However, we cannot yet predict gene transfer processes in soil to assess the frequency of unintentional transfer of engineered DNA to environmental microbes when applying synthetic biology technologies at scale. This challenge exists because of the complex and heterogeneous characteristics of soils, which contribute to the fitness and transport of cells and the exchange of genetic material within communities. Here, we describe knowledge gaps about gene transfer across soil microbiomes. Here, we propose strategies to improve our understanding of gene transfer across soil communities, highlight the need to benchmark the performance of biocontainment measures in situ, and discuss responsibly engaging community stakeholders. We highlight opportunities to address knowledge gaps, such as creating a set of soil standards for studying gene transfer across diverse soil types and measuring gene transfer host range across microbiomes using emerging technologies. By comparing gene transfer rates, host range, and persistence of engineered microbes across different soils, we posit that community-scale, environment-specific models can be built that anticipate biotechnology risks. Such studies will enable the design of safer biotechnologies that allow us to realize the benefits of synthetic biology and mitigate risks associated with the release of such technologies.

bioccontainment↗