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At least 91 records · Page 5

Bioreactor Development for CO2-Based In Situ Resource Utilization Manufacturing

Sustainable long-duration manned missions on both the Moon and Mars will require in situ resource utilization (ISRU). Carbon dioxide (CO2) has great potential as a harvestable resource, making up 95% of the atmosphere on Mars and being produced as respiratory waste in spacecraft and future planetary habitats. Through ISRU, biomanufacturing has the capability to produce a near limitless array of products from local space resources, which include pharmaceuticals, bioplastics, chemical feedstocks, and industrial enzymes. Here, a CO2-based ISRU recombinant protein bioreactor and associated biomanufacturing organisms were designed to produce a highly stable carbonic anhydrase (CA). Initial work characterized candidate organisms for growth on acetate and formic acid, carbon substrates that can be synthesized via electrochemical conversion of CO2. To improve growth on the CO2 producing substrate formic acid and for direct integration of ISRU CO2, a synthetic Calvin-Benson-Bassam cycle was designed for use in Cyberlindnera jadinii and Escherichia coli. Genetic modifications in E. coli will be facilitated by a tailored CRISPR/Cas9 and λ red recombineering two-vector system. For expression of CA, a blue light regulated T7 promoter was employed for dynamic and small molecule free induction. Efficient bioproduction through a fed-batch exponential feeding strategy was determined via mass balance calculations from ISRU substrates to biomass and CA yield. Flux balance analysis was used to model ISRU substrate metabolism and metabolic pathway engineering in candidate organisms under cultivation strategy conditions for both metabolism reconstruction and pathway design optimization. Finally, a small-scale, disposable bag bioreactor concept for use in the NASA Bioculture System infrastructure was designed to enable CO2-based CA production in reduced-gravity environments.

Biomanufacturing, Pathway Engineering, Flux Balanc↗

Bioreactor Development for CO2-Based In Situ Resource Utilization Manufacturing

Sustainable long-duration manned missions on both the Moon and Mars will require in situ resource utilization (ISRU). Carbon dioxide (CO2) has great potential as a harvestable resource, making up 95% of the atmosphere on Mars and being produced as respiratory waste in spacecraft and future planetary habitats. Through ISRU, biomanufacturing has the capability to produce a near limitless array of products from local space resources. Here, a CO2-based ISRU recombinant protein producing bioreactor and associated biomanufacturing organisms were designed to produce a highly stable carbonic anhydrase (CA). Candidate organisms were selected by growth characterization on acetate and formic acid, carbon substrates that are synthesized via electrochemical conversion of CO2. To improve growth on the CO2 producing substrate formic acid and for direct integration of ISRU CO2, a synthetic Calvin-Benson-Bassam cycle was designed for use in Escherichia coli. Multiplex genetic modification in E. coli was facilitated by a tailored CRISPR/Cas9 and λ red recombineering two-vector system. For expression of CA, a blue light regulated T7 promoter was employed for dynamic and small molecule free induction. Efficient bioproduction through a fed-batch exponential feeding strategy was determined via mass balance analysis from ISRU substrates to biomass and CA yield. Flux balance analysis was used to model ISRU substrate metabolism and metabolic pathway engineering in candidate organisms under cultivation strategy conditions for both metabolism reconstruction and pathway design optimization. Finally, a small-scale, disposable bag bioreactor for use in the NASA Bioculture System infrastructure was designed to enable CO2-based CA biomanufacturing in reduced-gravity environments.

biomanufacturing↗

Update to Developing a Multi-bioreactor Test Stand for the Evaluation of Biofilm Mitigation Technologies

Biofilm formation in spacecraft wastewater systems is a complex and impactful problem. Biomass flowing in these wastewater systems can cause clogging, higher differential pressures, and flow restrictions impacting equipment reliability and function. To explore this issue, the Marshall Space Flight Center’s biofilm test stand was created and has been utilized to examine multiple biofilm mitigation techniques over a long duration test time. This paper is an update to a previous paper, ICES-2021-122, and will serve to discuss the modifications and enhancements to the biofilm test stand. While the previous configuration tested heat treatments and coatings, this phase of testing will examine differing materials, competing bacteria provided by an external bioreactor, a prototype phytoremediation unit, and additional testing with a nutrient filter. Configuration changes include the addition of float switches, adjustment of the drain assembly architecture and additional support hardware for the competing bacteria bioreactor and phytoremediation units.

ECLSS↗

Update to Developing a Multi-bioreactor Test Stand for the Evaluation of Biofilm Mitigation Technologies

Biofilm formation in spacecraft wastewater systems is a complex and impactful problem. Biomass flowing in these wastewater systems can cause clogging, higher differential pressures, and flow restrictions impacting equipment reliability and function. To explore this issue, the Marshall Space Flight Center’s biofilm test stand was created and has been utilized to examine multiple biofilm mitigation techniques over a long duration test time. This paper is an update to a previous paper, ICES-2021-122, and will serve to discuss the modifications and enhancements to the biofilm test stand. While the previous configuration tested heat treatments and coatings, this phase of testing will examine differing materials, competing bacteria provided by an external bioreactor, a prototype phytoremediation unit, and additional testing with a nutrient filter. Configuration changes include the addition of float switches, adjustment of the drain assembly architecture and additional support hardware for the competing bacteria bioreactor and phytoremediation units.

ECLSS↗

Update to Developing a Multi-bioreactor Test Stand for the Evaluation of Biofilm Mitigation Technologies

Biofilm formation in spacecraft wastewater systems is a complex and impactful problem. Biomass flowing in these wastewater systems can cause clogging, higher differential pressures, and flow restrictions impacting equipment reliability and function. To explore this issue, the Marshall Space Flight Center’s biofilm test stand was created and has been utilized to examine multiple biofilm mitigation techniques over a long duration test time. This paper is an update to a previous paper, ICES-2021-122, and will serve to discuss the modifications and enhancements to the biofilm test stand. While the previous configuration tested heat treatments and coatings, this phase of testing will examine differing materials, competing bacteria provided by an external bioreactor, a prototype phytoremediation unit, and additional testing with a nutrient filter. Configuration changes include the addition of float switches, adjustment of the drain assembly architecture and additional support hardware for the competing bacteria bioreactor and phytoremediation units.

ECLSS↗

The fluid dynamic and shear environment in the NASA/JSC rotating-wall perfused-vessel bioreactor

The rotating-wall perfused-vessel (RWPV) bioreactor, used for both microgravity and Earth-based cell science experiments, is characterized in terms of the fluid dynamic and fluid shear stress environment. A numerical model of the flow field is developed and verified with laser Doppler velocimeter measurements. The effects of changes in operating conditions, including rotation rates and fluid perfusion rates, are investigated with the numerical model. The operating conditions typically used for ground-based experiments (equal rotation of the inner and outer cylinders) leads to flow patterns with relatively poor mass distribution characteristics. Approximately 50% of the inlet-perfused fluid bypasses the bulk of the fluid volume and flows to the perfusion exit. For operating conditions typical in microgravity, small differential rotation rates between the inner and outer cylinders lead to greatly improved flow distribution patterns and very low fluid shear stress levels over a large percentage of the fluid volume. Differences in flow patterns for the different operating conditions are explored. Large differences in the hydrodynamic environments for operating conditions typical of true microgravity and ground-based "microgravity simulations" are demonstrated.

Bioreactors↗

A Prototype Early Planetary Organic Processor Assembly (OPA) Based on Dual-Stage Anaerobic Membrane Bioreactor (AnMBR) for Fecal and Food Waste Treatment and Resource Recovery

Long-duration, deep-space exploration and habitation missions demand robust and reliable technologies to ensure crew health, safety, and mission success. Local food production will be essential for crew nutrition and morale. However, at $10,000/lb, the payload costs and mass/volume limitations to transport and provide the necessary resources, including fertilizer, for an anticipated 30-month mission become challenging over time. For mission success and sustainability, the Environmental Control and Life Support System (ECLSS) of the near future will need to recover resources from all “waste” sources and be near-closed loop. Organic wastes (e.g., fecal and food) offer a renewable source of C, N, P, water and other trace elements necessary to sustain crop production. However, these high solidwastes are often difficult to treatdue to factors includingheterogeneity, complexity, high organic strength, and the presence of pathogens. To date, there is no flight-ready technology capable of treating mixed organic wastes, creating a technology gap for future space missions. To address this need, a prototype Organic Processor Assembly (OPA) was developed through collaboration between the University of South Florida (USF) and NASA’s Kennedy Space Center (KSC). The OPA is based on the anaerobic membrane bioreactor (AnMBR), a hybrid technology coupling high-rate anaerobic digestion with membrane filtration. The system is designed for an early planetary base (EPB) scenario to aid in closing the resource recovery loop and decreaseresupply dependence. This presentation discusses initial research pertaining to: 1) design challenges in maximizing hydraulic/organic throughputandReliability, Availability, Maintainability, and Safety(RAMS)while minimizingmass and volume; 2) create capabilities for treating simulated high solids waste under steady and non-steady state conditions; and 3) measuresolids performance parameter(s). Future research and development pertaining to further optimization on system operation, performance, and expanded treatment capabilities arepresented.

Bioreactor↗

Design concepts for bioreactors in space

Microbial food sources are becoming viable and more efficient alternatives to conventional food sources, especially in the context of closed ecological life support systems (CELSS) in space habitats. Two bioreactor design concepts presented represent two dissimilar approaches to grappling with the absence of gravity in space habitats and deserve to be tested for adoption as important components of the life support function aboard spacecraft, space stations and other extra-terrestrial habitats.

Seshan, P. K.↗

Cell culture experiments planned for the space bioreactor

Culturing of cells in a pilot-scale bioreactor remains to be done in microgravity. An approach is presented based on several studies of cell culture systems. Previous and current cell culture research in microgravity which is specifically directed towards development of a space bioprocess is described. Cell culture experiments planned for a microgravity sciences mission are described in abstract form.

Morrison, Dennis R.↗

Application of gain scheduling to the control of batch bioreactors

The implementation of control algorithms to batch bioreactors is often complicated by the inherent variations in process dynamics during the course of fermentation. Such a wide operating range may render the performance of fixed gain PID controllers unsatisfactory. In this work, a detailed study on the control of batch fermentation is performed. Furthermore, a simple batch controller design is proposed which incorporates the concept of gain-scheduling, a subclass of adaptive control, with oxygen uptake rate as an auxiliary variable. The control of oxygen tension in the biorector is used as a vehicle to convey the proposed idea, analysis and results. Simulation experiments indicate significant improvement in controller performance can be achieved by the proposed approach even in the presence of measurement noise.

Cardello, Ralph↗

Optimal feedback control of a bioreactor with a remote sensor

Sensors used to monitor bioreactor conditions directly often perform poorly in the face of adverse nonphysiological conditions. One way to circumvent this is to use a remote sensor block. However, such a configuration usually causes a significant time lag between measurements and the actual state values. Here, the problem of implementing feedback control strategies for such systems, described by nonlinear equations, is addressed. The problem is posed as an optimal control problem with a linear quadratic performance index. The linear control law so obtained is used to implement feedback. A global linearization technique as well as an expansion using Taylor series is used to linearize the nonlinear system, and the feedback is subsequently implemented.

Niranjan, S. C.↗

Multimembrane Bioreactor

Set of hydrophilic and hydrophobic membranes in bioreactor allows product of reaction to be separated, while nutrients fed to reacting cells and byproducts removed from them. Separation process requires no externally supplied energy; free energy of reaction sufficient. Membranes greatly increase productivity of metabolizing cells by continuously removing product and byproducts, which might otherwise inhibit reaction, and by continuously adding oxygen and organic nutrients.

Cho, Toohyon↗

Controlled-Turbulence Bioreactors

Two versions of bioreactor vessel provide steady supplies of oxygen and nutrients with little turbulence. Suspends cells in environment needed for sustenance and growth, while inflicting less damage from agitation and bubbling than do propeller-stirred reactors. Gentle environments in new reactors well suited to delicate mammalian cells. One reactor kept human kidney cells alive for as long as 11 days. Cells grow on carrier beads suspended in liquid culture medium that fills cylindrical housing. Rotating vanes - inside vessel but outside filter - gently circulates nutrient medium. Vessel stationary; magnetic clutch drives filter cylinder and vanes. Another reactor creates even less turbulence. Oxygen-permeable tubing wrapped around rod extending along central axis. Small external pump feeds oxygen to tubing through rotary coupling, and oxygen diffuses into liquid medium.

Wolf, David A.↗

Hydrodynamic effects on cell growth in agitated microcarrier bioreactors

The net growth rate of bovine embryonic kidney cells in microcarrier bioreactor is the result of a variable death rate imposed on a cell culture trying to grow at a constant intrinsic growth rate. The death rate is a function of the agitation conditions in the system, and increases at higher agitation because of increasingly energetic interactions of the cell covered microcarriers with turbulent eddies in the fluid. At very low agitation rates bead-bead bridging becomes important; the large clumps formed by bridging can interact with larger eddies than single beads, leading to a higher death rate at low agitation. The growth and death rate were correlated with a dimensionless eddy number which compares eddy forces to the buoyant force on the bead.

Cherry, Robert S.↗

An expert system based intelligent control scheme for space bioreactors

An expert system based intelligent control scheme is being developed for the effective control and full automation of bioreactor systems in space. The scheme developed will have the capability to capture information from various resources including heuristic information from process researchers and operators. The knowledge base of the expert system should contain enough expertise to perform on-line system identification and thus be able to adapt the controllers accordingly with minimal human supervision.

San, Ka-Yiu↗

Perfusion Bioreactor Module

Perfusion bioreactor module, self-contained, closed-loop cell-culture system that operates in microgravity or on Earth. Equipment supports growth or long-term maintenance of cultures of human or other fragile cells for experiments in basic cell biology or process technology. Designed to support proliferation (initially at exponential rates of growth) of cells in complex growth medium and to maintain confluent cells in defined medium under conditions optimized to permit or encourage selected functions of cells, including secretion of products of cells into medium.

Morrison, Dennis R.↗