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Hydrogen production from salt water by Marine blue green algae and solar radiation
Two marine bluegreen algae, Oscillatoria sp. Miami BG 7 and Synechococcus sp Miami 041511 have been selected as the result of over 10 years continuous and intensive effort of isolation, growth examination, and the screening of hydrogen photoproduction capability in this laboratory. Both strains photoproduced hydrogen for several days at high rates and a quantity of hydrogen was accumulated in a closed vessel. Overall hydrogen donor substance of the hydrogen photoproduction was found to be salt water. Using strain Miami BG 7, a two step method of hydrogen photoproduction from salt water was successfully developed and this was recycled several times over a one month period using both free cells and immobilized cells in both indoor and outdoor under natural sunlight. According to these experiments, a prototype floating hydrogen production system was designed for further development of the biosolar hydrogen production system.
Strategies to achieve high productivity, high conversion, and high yield in yeast fermentation of algal biomass hydrolysate
The conversion of carbohydrates in biomass via fermentation is an important component of an overall strategy to decarbonize the production of fuels and chemicals. Owing to the cost and resources required to produce biomass hydrolysates, the economic and environmental sustainability of these fermentation processes requires that they operate with high yields, sugar conversion, and productivity. Immobilized-cell technology in a continuous bioprocess can achieve significantly higher volumetric productivities than is possible from standard batch fermentation using free cells. Here, we demonstrate approaches for improvement of ethanol yield from algal hydrolysates and a mock hydrolysate medium. Saccharomyces cerevisiae was immobilized in alginate and incorporated into a two-column immobilized cell reactor system. Furthermore, the yeast quorum-sensing molecule, 2-phenylethanol, was added to improve ethanol yield by restricting growth and diverting sugar to ethanol. The bioreactor system could achieve high ethanol volumetric productivity (>20 g/ Lreactor ·h) and high glucose conversion (>99%) in mock hydrolysate, while the addition of 0.2% 2-phenylethanol resulted in 4.9% higher ethanol yield. With an algal hydrolysate of <10 g/L sugar, the ethanol volumetric productivity reached 9.8 g/ Lreactor ·h, and the addition of 0.2% 2-phenylethanol increased the ethanol yield by up to 7.4%. These results demonstrate the feasibility of novel strategies to achieve sustainability goals in biomass conversions.
A bioreactor system for the nitrogen loop in a Controlled Ecological Life Support System
As space missions become longer in duration, the need to recycle waste into useful compounds rises dramatically. This problem can be addressed by the development of Controlled Ecological Life Support Systems (CELSS) (i.e., Engineered Closed/Controlled Eco-Systems (ECCES)), consisting of human and plant modules. One of the waste streams leaving the human module is urine. In addition to the reclamation of water from urine, recovery of the nitrogen is important because it is an essential nutrient for the plant module. A 3-step biological process for the recycling of nitrogenous waste (urea) is proposed. A packed-bed bioreactor system for this purpose was modeled, and the issues of reaction step segregation, reactor type and volume, support particle size, and pressure drop were addressed. Based on minimization of volume, a bioreactor system consisting of a plug flow immobilized urease reactor, a completely mixed flow immobilized cell reactor to convert ammonia to nitrite, and a plug flow immobilized cell reactor to produce nitrate from nitrite is recommended. It is apparent that this 3-step bioprocess meets the requirements for space applications.
Ex Vivo Growth of Bioengineered Ligaments and Other Tissues
A method of growing bioengineered tissues for use in surgical replacement of damaged anterior cruciate ligaments has been invented. An anterior cruciate ligament is one of two ligaments (the other being the posterior cruciate ligament) that cross in the middle of a knee joint and act to prevent the bones in the knee from sliding forward and backward relative to each other. Anterior cruciate ligaments are frequently torn in sports injuries and traffic accidents, resulting in pain and severe limitations on mobility. By making it possible to grow replacement anterior cruciate ligaments that structurally and functionally resemble natural ones more closely than do totally synthetic replacements, the method could create new opportunities for full or nearly full restoration of functionality in injured knees. The method is also adaptable to the growth of bioengineered replacements for other ligaments (e.g., other knee ligaments as well as those in the hands, wrists, and elbows) and to the production of tissues other than ligaments, including cartilage, bones, muscles, and blood vessels. The method is based on the finding that the histomorphological properties of a bioengineered tissue grown in vitro from pluripotent cells within a matrix are affected by the direct application of mechanical force to the matrix during growth generation. This finding provides important new insights into the relationships among mechanical stress, biochemical and cell-immobilization methods, and cell differentiation, and is applicable to the production of the variety of tissues mentioned above. Moreover, this finding can be generalized to nonmechanical (e.g., chemical and electromagnetic) stimuli that are experienced in vivo by tissues of interest and, hence, the method can be modified to incorporate such stimuli in the ex vivo growth of replacements for the various tissues mentioned above. In this method, a three-dimensional matrix made of a suitable material is seeded with pluripotent stem cells. The patient s bone-marrow stromal cells are preferably used as the pluripotent cells in this method. Suitable matrix materials are materials to which cells can adhere for example, collagen type I. The seeded matrix is attached to anchors at opposite ends and then the cells in the matrix are cultured under conditions appropriate for the growth and regeneration of cells. Suitable anchor materials are materials to which the matrix can attach; examples include demineralized bone and Goinopra coral that has been treated to convert its calcium carbonate to calcium phosphate.
Synthesis and characterization of poly/glutaraldehyde/ - A potential reagent for protein immobilization and cell separation
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Analysis of cell performance and thermal regeneration of a lithium-tin cell having an immobilized fused-salt electrolyte
Cell performance and thermal regeneration of a thermally regenerative cell uses lithium and tin and a fused-salt electrolyte. The emf of the Li-Sn cell, as a function of cathode-alloy composition, is shown to resemble that of the Na-Bi cell.
Sol–Gel‐Based Advanced Porous Silica Materials for Biomedical Applications
Abstract Porous silica‐based materials have burgeoning applications ranging from fillers and additives, to adsorbents, catalysts, and recently therapeutic agents and vaccines in nanomedicine. The preponderance of these materials is made by sol–gel processing wherein soluble silica precursors are reacted to form amorphous networks composed of siloxane bonds. The facile sol–gel approach allows for an unlimited variety of binary tertiary and more complex chemical compositions including organic ligands and networks resulting in so‐called organic–inorganic hybrid materials. Here, a brief review of the recent progress in sol–gel‐derived silica materials prepared as particles, thin films, biosilica/silica bioreplicas (of molecules, cells and organisms), and their related preparation, properties, and bioapplications is provided. First, it highlights the recent achievements of mesoporous silica nanoparticles in biomedical applications, including therapeutic agent delivery, multimodal imaging and theranostics, and bone tissue engineering and repair. Second, the research in evaporation‐induced self‐assembly (EISA)‐based mesostructured silica thin films and cell‐directed EISA in bio/nano interfaces for various bioapplications, such as bioactive coatings, biosensing, and living cell immobilization, has been reviewed. Third, the pioneering work in biomimetic silicification/immobilization of biomolecules and bio‐organisms and silica bioreplication of complex bio‐organisms is summarized. Finally, it is concluded with personal perspectives on the directions of future work on this field.
Microbe-Encapsulated Silica Gel Biosorbents for Selective Extraction of Scandium from Coal Byproducts
Establishing an inexpensive and environmentally friendly scandium (Sc) supply is critical for the development of clean energy technologies. However, refining Sc from Sc-bearing sources using current technologies poses economic and environmental challenges due to its low concentration relative to base metals and chemically similar lanthanides. In this work, we developed a biosorption-based flow-through process for extraction of Sc from low-grade feedstocks. A novel biosorbent material was synthesized by encapsulating Arthrobacter nicotianae, a bacterium that selectively adsorbs Sc, within a porous silica matrix. Through the use of several complementary microscopy techniques, we demonstrated a highly porous biosorbent structure with a high cell-loading density and a homogenous cell distribution. Batch adsorption assays revealed selective Sc adsorption over lanthanides and common based metals, with the exception of Fe(III), as evidenced by separation factors greater than 50. The MESG particles were packed into fixed-bed columns to enable adsorption under flow-through conditions, which demonstrated effective Sc extraction at flow rates up to 0.08 cm/s and high stability for reuse; greater than 95% of the adsorption capacity was maintained after 10 consecutive adsorption/desorption cycles. When applied to a lignite coal leachate, whose Fe content was depleted through pH-mediated precipitation, the MESG particles yielded Sc breakthrough at 30 bed volumes, whereas all other metals broke through after only a few bed volumes. Following desorption, an eluate with a 124-fold increase in Sc purity was achieved relative to the lignite leachate with Sc constituting 96.4% of the total REEs. This study established a rapid, facile, and scalable cell immobilization approach that enables the use of microbial biomass for Sc recovery from low-grade sources.
Spatial distribution of total, ammonia-oxidizing, and denitrifying bacteria in biological wastewater treatment reactors for bioregenerative life support
Bioregenerative life support systems may be necessary for long-term space missions due to the high cost of lifting supplies and equipment into orbit. In this study, we investigated two biological wastewater treatment reactors designed to recover potable water for a spacefaring crew being tested at Johnson Space Center. The experiment (Lunar-Mars Life Support Test Project-Phase III) consisted of four crew members confined in a test chamber for 91 days. In order to recycle all water during the experiment, an immobilized cell bioreactor (ICB) was employed for organic carbon removal and a trickling filter bioreactor (TFB) was utilized for ammonia removal, followed by physical-chemical treatment. In this study, the spatial distribution of various microorganisms within each bioreactor was analyzed by using biofilm samples taken from four locations in the ICB and three locations in the TFB. Three target genes were used for characterization of bacteria: the 16S rRNA gene for the total bacterial community, the ammonia monooxygenase (amoA) gene for ammonia-oxidizing bacteria, and the nitrous oxide reductase (nosZ) gene for denitrifying bacteria. A combination of terminal restriction fragment length polymorphism (T-RFLP), sequence, and phylogenetic analyses indicated that the microbial community composition in the ICB and the TFB consisted mainly of Proteobacteria, low-G+C gram-positive bacteria, and a Cytophaga-Flexibacter-Bacteroides group. Fifty-seven novel 16S rRNA genes, 8 novel amoA genes, and 12 new nosZ genes were identified in this study. Temporal shifts in the species composition of total bacteria in both the ICB and the TFB and ammonia-oxidizing and denitrifying bacteria in the TFB were also detected when the biofilms were compared with the inocula after 91 days. This result suggests that specific microbial populations were either brought in by the crew or enriched in the reactors during the course of operation.
Development of a gravity-independent wastewater bioprocessor for advanced life support in space
Operation of aerobic biological reactors in space is controlled by a number of challenging constraints, mainly stemming from mass transfer limitations and phase separation. Immobilized-cell packed-bed bioreactors, specially designed to function in the absence of gravity, offer a viable solution for the treatment of gray water generated in space stations and spacecrafts. A novel gravity-independent wastewater biological processor, capable of carbon oxidation and nitrification of high-strength aqueous waste streams, is presented. The system, consisting of a fully saturated pressurized packed bed and a membrane oxygenation module attached to an external recirculation loop, operated continuously for over one year. The system attained high carbon oxidation efficiencies often exceeding 90% and ammonia oxidation reaching approximately 60%. The oxygen supply module relies on hydrophobic, nonporous, oxygen selective membranes, in a shell and tube configuration, for transferring oxygen to the packed bed, while keeping the gaseous and liquid phases separated. This reactor configuration and operating mode render the system gravity-independent and suitable for space applications.
Closure of Regenerative Life Support Systems: Results of the Lunar-Mars Life Support Test Project
Future long duration human exploration missions away from Earth will require closed-loop regenerative life support systems to reduce launch mass, reduce dependency on resupply and increase the level of mission self sufficiency. Such systems may be based on the integration of biological and physiocochemical processes to produce potable water, breathable atmosphere and nutritious food from metabolic and other mission wastes. Over the period 1995 to 1998 a series of ground-based tests were conducted at the National Aeronautics and Space Administration, Johnson Space Center, to evaluate the performance of advanced closed-loop life support technologies with real human metabolic and hygiene loads. Named the Lunar-Mars Life Support Test Project (LMLSTP), four integrated human tests were conducted with increasing duration, complexity and closure. The first test, LMLSTP Phase I, was designed to demonstrate the ability of higher plants to revitalize cabin atmosphere. A single crew member spent 15 days within an atmospherically closed chamber containing 11.2 square meters of actively growing wheat. Atmospheric carbon dioxide and oxygen levels were maintained by control of the rate of photosynthesis through manipulation of light intensity or the availability of carbon dioxide and included integrated physicochemical systems. During the second and third tests, LMLSTP Phases II & IIa, four crew members spent 30 days and 60 days, respectively, in a larger sealed chamber. Advanced physicochemical life support hardware was used to regenerate the atmosphere and produce potable water from wastewater. Air revitalization was accomplished by using a molecular sieve and a Sabatier processor for carbon dioxide absorption and reduction, respectively, with oxygen generation performed by water hydrolysis. Production of potable water from wastewater included urine treatment (vapor compression distillation), primary treatment (ultrafiltration/reverse osmosis and multi-filtration) and post processing. For the Phase II test the water recovery rate ranged from 95 to 98%, depending on the processor. LMLSTP Phase III, the fourth test of the series, had a duration of 91 days and included four crew members. The test demonstrated an integration of physicochemical and biological technologies for air revitalization, water recovery and waste processing. Wheat supplemented the physicochemical air revitalization systems by providing approximately 25% of the oxygen required for the 4-person crew. The water recovery system included immobilized cell and trickling filter bioreactors for primary water treatment, reverse osmosis and air evaporation systems for secondary water treatment, followed by post processing. The 8 day initial supply of water was recycled through the chamber and crew 10 times over the course of the test. Grain from the wheat together with fresh lettuce from a small growth chamber within the crew chamber provided supplementation to the stored food system, but at a level less than 5% of the crew s caloric requirement. An incinerator was used to demonstrate mineralization of the crew s solid waste, with the combustion products (mainly carbon dioxide) returned to the wheat for conversion to oxygen.
Culture systems and methods of using same
Culture systems and methods of using same. The systems include a housing defining an inner space. The inner space includes a headspace and at least a portion of a reservoir. A surface for immobilizing cells is moveable between the headspace and the reservoir. The systems can be used for coculturing methanotrophs and phototrophs for processing biogas and wastewater, particularly from anaerobic digesters.
Culture systems and methods of using same
Culture systems and methods of using same. The systems include a housing defining an inner space. The inner space includes a headspace and at least a portion of a reservoir. A surface for immobilizing cells is moveable between the headspace and the reservoir. The systems can be used for coculturing methanotrophs and phototrophs for processing biogas and wastewater, particularly from anaerobic digesters.
Durable Fuel Cell MEA through Immobilization of Catalyst Particle and Membrane Chemical Stabilizer
For hydrogen-based fuel cell electric vehicle to be cost competitive with the incumbent diesel-powered internal combustion engine heavy-duty vehicle, the total cost of ownership of the truck must be decreased via both capital cost and the operating cost (H 2 fuel expense) reduction. This puts the emphasis on the decrease of fuel cell stack cost via use of low platinum catalyst material usage to decrease capital cost, high H 2 fuel efficiency and high stack durability to decrease the operating cost. US DOE has set a target of ≤ 0.25 mgPt/cm 2 total loading in the membrane electrode assembly, high fuel efficiency of > 68% and a HD-combined target of ≥ 2.5 kW/gPGM after running an accelerated stress test (AST) equivalent to 30,000 hours of heavy-duty fuel cell operation.
Cell Free and Immobilization Technologies (CFIT)
Today, several key factors negatively impact the production of fuels and chemicals from renewable sources. Common hindrances in the biological production of biochemicals are: (1) end-product or intermediate toxicity to the microbial biocatalyst, (2) the diversion of carbon to biomass formation, and (3) co-production of undesired byproducts. A particularly attractive alternative is to eliminate the biocatalyst entirely and instead operate the desired metabolic pathways in isolation, thus circumventing the roadblocks of biological toxicity, lower yields, and lack of specificity. However, cell-free enzyme systems still suffer from low productivities owing in part to the effects of free diffusion of intermediates, lack of long term enzyme stability, cofactor cost or inefficient recycling rates, and finally, the cost of enzyme production/purification. This project represents a new effort to propose innovative and cost competitive routes to producing biochemicals from a variety of feedstocks using cell free approaches. These routes will help reduce the current risk and cost associated with classical cell free production. Cell free technologies show promise for application to the production of toxic/inhibitory products or products difficult to separate from microbial growth media and can help reduce the production barriers in multiple areas of biological conversion of feedstocks to biochemicals. More specifically, we are developing new technologies and routes that could be used to produce high value biochemicals such as 2,3 BDO and terpenes (among many others) from biomass derived C5/C6 sugars or lignin but also from waste byproducts such as glycerol. This project will lead to significant innovation and also lead to new concepts and rational design of pathways and enzymes. Within this project, we will develop new metabolic enzyme cascades that will represent natural or artificial combinations of enzymes to produce the desired biochemicals from a variety of feedstocks. We are also developing basic design principals for constructing synthetic metabolons, using fusion proteins and synthetic protein scaffolds, to promote substrate channeling and stability while conserving peak activity. Additionally, our efforts include a techno economic analysis (TEA) of cell free approaches to provide the sensitivities of the process to enzyme loading, activity, pH, reactor volumes, cofactor recycling rates. Finally, we are focusing on further increasing stability, operating lifetime and efficiency of the pathway enzymes by immobilization on support surfaces. We are also focus on immobilizing pathway enzymes or combinations of enzymes on several different conducting polymers and evaluate the effect on stability and operating lifetime. As more combinations become available we will conduct a more systematic study of the means of immobilizing these enzymes. This preliminary work will enable the in-depth study of cofactor recycling at these interfaces using mediators for electron transfer. Taken together, these approaches will enable process intensification, continuous operation, lower capital and separations costs and end-product flexibility, and thus has the potential to contribute significantly to BETO's goals of cost competitive biofuels and bioproducts. To date we have demonstrated 1) the conversion of pyruvate to 2,3 BDO (4 enzymes with cofactor recycling, no additional cofactors needed) at >100g/L (>3g/L/h) using our enzyme tethering approach, 2) Generated mutants of a key redox enzyme with >180- fold improvement in NADH utilization over WT to >85% of NADPH utilization. 3) Produced mevalonate at >10g/L from glucose, limonene at >10g/L from mevalonate and more than >5g/L from glucose with crude enzyme preparations and complete cofactor recycling. 4) successfully generated fully active cross-linked aggregates (CLEAs) of NOX, a cofactor-regenerating water-forming oxidase that is key to our process with increased stability 5) Engineered and identified enzymes able to use the biomimetic cofactor NMN and synthesized new synthetic cofactors.
BETO 2021 Peer Review - Cell Free and Immobilization Technologies (CFIT)
Today, several key factors negatively impact the production of fuels and chemicals from renewable sources. Common hindrances in the biological production of biochemicals are: (1) end-product or intermediate toxicity to the microbial biocatalyst, (2) the diversion of carbon to biomass formation, and (3) co-production of undesired byproducts. A particularly attractive alternative is to eliminate the biocatalyst entirely and instead operate the desired metabolic pathways in isolation, thus circumventing the roadblocks of biological toxicity, lower yields, and lack of specificity. However, cell-free enzyme systems still suffer from low productivities owing in part to the effects of free diffusion of intermediates, lack of long term enzyme stability, cofactor cost or inefficient recycling rates, and finally, the cost of enzyme production/purification. This project represents a new effort to propose innovative and cost competitive routes to producing biochemicals from a variety of feedstocks using cell free approaches. These routes will help reduce the current risk and cost associated with classical cell free production. Cell free technologies show promise for application to the production of toxic/inhibitory products or products difficult to separate from microbial growth media and can help reduce the production barriers in multiple areas of biological conversion of feedstocks to biochemicals.
Monolithic Continuous-Flow Bioreactors
Monolithic ceramic matrices containing many small flow passages useful as continuous-flow bioreactors. Ceramic matrix containing passages made by extruding and firing suitable ceramic. Pores in matrix provide attachment medium for film of cells and allow free movement of solution. Material one not toxic to micro-organisms grown in reactor. In reactor, liquid nutrients flow over, and liquid reaction products flow from, cell culture immobilized in one set of channels while oxygen flows to, and gaseous reaction products flow from, culture in adjacent set of passages. Cells live on inner surfaces containing flowing nutrient and in pores of walls of passages. Ready access to nutrients and oxygen in channels. They generate continuous high yield characteristic of immobilized cells, without large expenditure of energy otherwise incurred if necessary to pump nutrient solution through dense biomass as in bioreactors of other types.