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At least 19 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2015

Integrated ‘omics analysis for studying the microbial community response to a pH perturbation of a cellulose-degrading bioreactor culture

Propionate accumulation is a common indicator of process imbalances in anaerobic bioreactor systems. The accumulation of propionate can occur due to low retention rates, hydrogen accumulation, or mechanical changes affecting the proximity between propionate oxidizers and partner species, thereby preventing necessary electron transfer. Few studies, however, have observed the changes in microbial community structure during propionate accumulation. We used 454 pyrosequencing of 16S rDNA to evaluate the community membership during propionate accumulations in replicate bioreactors with rumen based cultures. Half of the culture volume from a parent reactor was transferred to a sterile “daughter” reactor, and both systems were run identically. Both reactors experienced a propionate accumulation after roughly 10 days, with the propionate accumulation being less pronounced in the parent reactor as compared to the daughter reactor. Non-metric multidimensional scaling (NMDS) was used to determine clustering patterns of the samples, and correlative methods were used to determine which OTUs were significantly associated with the movements of samples along the NMDS axes. The presence of Saccharofermentans characterized the position of early samples, whereas the presence of Ruminococcus and Succiniclasticum were more indicative of the positions of later samples. Hydrogen accumulation and low sequence counts indicated low methanogen activity. Although both reactor systems were closed to microbial inputs due to the sterilization of influent media, we recorded significant increases in reactor diversity over time. This suggests that changes in the abundances of dominant community members may affect the sequencing of rare taxa within samples.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Impacts of intentional mycoplasma contamination on CHO cell bioreactor cultures

Mycoplasma contamination events in biomanufacturing facilities can result in loss of production and costly cleanups. Mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and may penetrate the 0.2 µm filters often used in the primary clarification of harvested cell culture fluid. Culture cell-based and indicator cell-based assays that are used to detect mycoplasma are highly sensitive but can take up to 28 days to complete and cannot be used for real-time decision making during the biomanufacturing process. To support real-time measurements of mycoplasma contamination, there is a push to explore nucleic acid testing. However, cell-based methods measure growth or colony forming units and nucleic acid testing measures genome copy number; this has led to ambiguity regarding how to compare the sensitivity of the methods. In addition, the high risk of conducting experiments wherein one deliberately spikes mycoplasma into bioreactors has dissuaded commercial groups from performing studies to explore the multiple variables associated with the upstream effects of a mycoplasma contamination in a manufacturing setting. Here we studied the ability of Mycoplasma arginini to persist in a single-use, perfusion rocking bioreactor system containing a Chinese hamster ovary (CHO) DG44 cell line expressing a model monoclonal immunoglobulin G1 (IgG1) antibody. We examined M. arginini growth and detection by culture methods, as well as the effects of M. arginini on mammalian cell health, metabolism, and productivity. We compared process parameters and controls normally measured in bioreactors including dissolved oxygen, gas mix, and base addition to maintain pH, to examine parameter changes as potential indicators of contamination. Our work showed that M. arginini affects CHO cell growth profile, viability, nutrient consumption, oxygen use, and waste production at varying timepoints after M. arginini introduction to the culture. Importantly, how the M. arginini contamination impacts the CHO cells is influenced by the concentration of CHO cells and rate of perfusion at the time of M. arginini spike. Careful evaluation of dissolved oxygen, pH control parameters, ammonia, and arginine over time may be used to indicate Mycoplasma contamination events in biomanufacturing facilities can result in loss of production and costly cleanups. Mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and may penetrate the 0.2 µm filters often used in the primary clarification of harvested cell culture fluid. Culture cell-based and indicator cell-based assays that are used to detect mycoplasma are highly sensitive but can take up to 28 days to complete and cannot be used for real-time decision making during the biomanufacturing process. To support real-time measurements of mycoplasma contamination, there is a push to explore nucleic acid testing. However, cell-based methods measure growth or colony forming units and nucleic acid testing measures genome copy number; this has led to ambiguity regarding how to compare the sensitivity of the methods. In addition, the high risk of conducting experiments wherein one deliberately spikes mycoplasma into bioreactors has dissuaded commercial groups from performing studies to explore the multiple variables associated with the upstream effects of a mycoplasma contamination in a manufacturing setting. Here we studied the ability of Mycoplasma arginini to persist in a single-use, perfusion rocking bioreactor system containing a Chinese hamster ovary (CHO) DG44 cell line expressing a model monoclonal immunoglobulin G1 (IgG1) antibody. We examined M. arginini growth and detection by culture methods, as well as the effects of M. arginini on mammalian cell health, metabolism, and productivity. We compared process parameters and controls normally measured in bioreactors including dissolved oxygen, gas mix, and base addition to maintain pH, to examine parameter changes as potential indicators of contamination. Our work showed that M. arginini affects CHO cell growth profile, viability, nutrient consumption, oxygen use, and waste production at varying timepoints after M. arginini introduction to the culture. Importantly, how the M. arginini contamination impacts the CHO cells is influenced by the concentration of CHO cells and rate of perfusion at the time of M. arginini spike. Careful evaluation of dissolved oxygen, pH control parameters, ammonia, and arginine over time may be used to indicate Mycoplasma contamination events in biomanufacturing facilities can result in loss of production and costly cleanups. Mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and may penetrate the 0.2 µm filters often used in the primary clarification of harvested cell culture fluid. Culture cell-based and indicator cell-based assays that are used to detect mycoplasma are highly sensitive but can take up to 28 days to complete and cannot be used for real-time decision making during the biomanufacturing process. To support real-time measurements of mycoplasma contamination, there is a push to explore nucleic acid testing. However, cell-based methods measure growth or colony forming units and nucleic acid testing measures genome copy number; this has led to ambiguity regarding how to compare the sensitivity of the methods. In addition, the high risk of conducting experiments wherein one deliberately spikes mycoplasma into bioreactors has dissuaded commercial groups from performing studies to explore the multiple variables associated with the upstream effects of a mycoplasma contamination in a manufacturing setting. Here we studied the ability of Mycoplasma arginini to persist in a single-use, perfusion rocking bioreactor system containing a Chinese hamster ovary (CHO) DG44 cell line expressing a model monoclonal immunoglobulin G1 (IgG1) antibody. We examined M. arginini growth and detection by culture methods, as well as the effects of M. arginini on mammalian cell health, metabolism, and productivity. We compared process parameters and controls normally measured in bioreactors including dissolved oxygen, gas mix, and base addition to maintain pH, to examine parameter changes as potential indicators of contamination. Our work showed that M. arginini affects CHO cell growth profile, viability, nutrient consumption, oxygen use, and waste production at varying timepoints after M. arginini introduction to the culture. Importantly, how the M. arginini contamination impacts the CHO cells is influenced by the concentration of CHO cells and rate of perfusion at the time of M. arginini spike. Careful evaluation of dissolved oxygen, pH control parameters, ammonia, and arginine over time may be used to indicate mycoplasma contamination in CHO cell cultures in a bioreactor before a read-out from a traditional method.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2019

Design, Prototyping, and Testing of a Novel Flowpath with an Array of Six 3D Matrix Vitvo Bioreactors for the NASA Bioculture System

The NASA Bioculture System is an advanced cell culture closed-loop system containing highly automated flowpaths designed to conduct long term biology experiments on ISS with earth remote controllable medium flow, temperature, gas composition, medium exchange, cell sampling and fixation. This technology was already demonstrated with successful cardiomyocyte and osteocyte cultures experiments onboard the ISS and is now supporting NASA PI science. The Bioculture System, however, can only support 10 cassettes with disposable flowpaths, each containing a single hollow fiber bioreactor with a culture capacity of about 2ml. This constraint not only severely limits the number of investigators that can conduct experiments in space, but also subjects the experiments to limitations in the number of replicates and conditions that can be studied. To address these limitations, we sought a novel design solution to maximize the number of separate bioreactor cultures and volume that can be conducted simultaneously. To this end we designed, prototyped, and are now testing a six-Vitvo 3D Matrix 2ml bioreactor insert that replaces the conventional Bioculture System hollow fiber bioreactor. This design will allow the Bioculture System to support up to 60 different bioreactors and samples at once. Specifically, the novel gas-tight containment housing insert contains six COTS Rigenerand VITVO bioreactors stacked on each side of a heat sink powered by the existing heating element and pair of temperature sensors. Medium will be distributed into each bioreactor's cell-free chamber via its built-in Luer connector, then across the 3D matrix to the cell chamber, dissipating laminar flow and limiting fluid shear stresses that might mechanostimulate cell cultures. Gas (5% CO2 in air) will be supplied directly to the bioreactor gas-tight housing for exchange via the bioreactor flat-surface gas-permeable membranes, eliminating the need for the existing Bioculture System cassette oxygenator. If successfully implemented on ISS, this new multi-bioreactor insert for the Bioculture System has the potential to make real-time cell science experimentation in space more efficient and accessible to more investigators.

Ma, Kristin R.↗
NASA NTRS2002

An Optical Oxygen Sensor for Long-Term Continuous Monitoring of Dissolved Oxygen in Perfused Bioreactors

For long-term growth of man1ITlalian cells in perfused bioreactors, it is essential to monitor the concentration of dissolved oxygen (DO) present in the culture medium to quantitate and control level of DO. Continuous measurement of the amount of DO in the cell culture medium in-line under sterile conditions in NASA's perfused bioreactor requires that the oxygen sensor provide increased sensitivity and be sterilizable and nontoxic. Additionally, long-term cell culture experiments require that the calibration be maintained several weeks or months. Although there are a number of sensors for dissolved oxygen on the market and under development elsewhere, very few meet these stringent conditions. An optical oxygen sensor (BOXY) based on dynamic fluorescent quenching and a pulsed blue LED light source was developed in our laboratory to address these requirements. Tris( 4,7 -diphenyl-l, 1 O-phenanthroline )ruthenium(II) chloride is employed as the fluorescent dye indicator. The sensing element consists of a glass capillary (OD 4.0 mm; ID 2.0 mm) coated internally with a thin layer of the fluorescent dye in silicone matrix and overlayed with a black shielding layer. Irradiation of the sensing element with blue light (blue LED with emission maximum at 475 nm) generates a red fluorescence centered at 626 nm. The fluorescence intensity is correlated to the concentration of DO present in the culture medium, following the modified non-linear Stern-Volmer equation. By using a pulsed irradiating light source, the problem of dye-bleaching, which is often encountered in long-term continuous measurements of tIns type, 'is minimized. To date we achieved sensor resolution of 0.3 mmHg at 50 mmHg p02, and 0.6 mmHg at 100 mmHg p02, with a response time of about one minute. Calibration was accomplished in sterile phosphate-buffered saline with a blood-gas analyzer (BGA) measurement as reference. Stand-alone software was also developed to control the sensor and bioreactor as well as to acquire data. Two HOXY sensors with a single calibration were employed to continuously monitor the DO in GTSF-2 medium during a Baby Hamster Kidney (BHK-21) cell culture in a Rotating Wall Perfused Vessel (RWPV) bioreactor for 90 days. HOXY sensors were located at the inlet to and outlet from the bioreactor. One of the sensors was placed between an oxygenator and the inlet to the bioreactor. The dissolved oxygen concentrations determined by both sensors were compared with those measured regularly with the BGA reference. The cell culture was maintained for 110 days. Sensor output was found to correlate well with the BGA data throughout the experiment, where the DO of the medium ranged between 25 and 50 mmHg at the bioreactor outlet and 90-130 mmHg at the bioreactor inlet. Measuring DO with the HOXY sensors versus the BGA reference indicated bias values of -2 mmHg and -15 mmHg, and precision values of +/-3mmHg and +/-16 mmHg at the bioreactor inlet and outlet, respectively.

Gao, F. G.↗
DOE OSTI2015

A comparative study of leachate quality and biogas generation in simulated anaerobic and hybrid bioreactors

Highlights: • Temporary aeration shortened the initial acid inhibition phase for methanogens. • COD decreased faster in the hybrid bioreactor than that in the anaerobic control. • Methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. • MSW settlement increased with increasing the frequency of intermittent aeration. - Abstract: Research has been conducted to compare leachate characterization and biogas generation in simulated anaerobic and hybrid bioreactor landfills with typical Chinese municipal solid waste (MSW). Three laboratory-scale reactors, an anaerobic (A1) and two hybrid bioreactors (C1 and C2), were constructed and operated for about 10 months. The hybrid bioreactors were operated in an aerobic–anaerobic mode with different aeration frequencies by providing air into the upper layer of waste. Results showed that the temporary aeration into the upper layer aided methane generation by shortening the initial acidogenic phase because of volatile fatty acids (VFAs) reduction and pH increase. Chemical oxygen demand (COD) decreased faster in the hybrid bioreactors, but the concentrations of ammonia–nitrogen in the hybrid bioreactors were greater than those in the anaerobic control. Methanogenic conditions were established within 75 d and 60 d in C1 and C2, respectively. However, high aeration frequency led to the consumption of organic matters by aerobic degradation and resulted in reducing accumulative methane volume. The temporary aeration enhanced waste settlement and the settlement increased with increasing the frequency of aeration. Methane production was inhibited in the anaerobic control; however, the total methane generations from hybrid bioreactors were 133.4 L/kg{sub vs} and 113.2 L/kg{sub vs}. As for MSW with high content of food waste, leachate recirculation right after aeration stopped was not recommended due to VFA inhibition for methanogens.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2022

Bioreactors: Design, Background, and Applications

A bioreactor is any reaction vessel designed to contain a biological reaction with the aims of either growing an organism, producing a biological product, bioremediating waste, or investigating the workings of a biological process within a controlled environment. Bioreactors compose a broad category of useful and ubiquitous technologies present throughout the bioeconomy, which are inherently “dualuse” technologies: holding both immense potential for both benefit and harm to society. Due to this duality, national security professionals must bridge technological understanding and policy acumen to understand how bioreactors are integrated into both, many peaceful applications throughout the global bioeconomy such as in developing or manufacturing life-saving pharmaceuticals, as well as potentially harmful and/or illicit uses, such as producing biowarfare agent. The following document gives a surface-level introduction into this broad category of technology. The first two sections of this document describe the historical origins, then typical contemporary uses of bioreactors as a critical component of the modern bioeconomy providing examples for each use case. The following two sections begin to give a technical description of bioreactor technology introducing the features commonly found in simple bioreactors and providing a non-exhaustive framework for categorizing bioreactors commonly found in wet-microbiology applications. The last two sections present two short discussions on the challenges facing modern bioreactor technology, firstly from a technical standpoint, and then from the non-technical standpoint with special attention to the dual-use dilemma and issue of responsible technological development inherent in most biotechnology.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1989

Design challenges for space bioreactors

The design of bioreactors for operation under conditions of microgravity presents problems and challenges. Absence of a significant body force such as gravity can have profound consequences for interfacial phenomena. Marangoni convection can no longer be overlooked. Many speculations on the advantages and benefits of microgravity can be found in the literature. Initial bioreactor research considerations for space applications had little regard for the suitability of the designs for conditions of microgravity. Bioreactors can be classified in terms of their function and type of operation. The complex interaction of parameters leading to optimal design and operation of a bioreactor is illustrated by the JSC mammalian cell culture system. The design of a bioreactor is strongly dependent upon its intended use as a production unit for cell mass and/or biologicals or as a research reactor for the study of cell growth and function. Therefore a variety of bioreactor configurations are presented in rapid summary. Following this, a rationale is presented for not attempting to derive key design parameters such as the oxygen transfer coefficient from ground-based data. A set of themes/objectives for flight experiments to develop the expertise for design of space bioreactors is then proposed for discussion. These experiments, carried out systematically, will provide a database from which engineering tools for space bioreactor design will be derived.

Seshan, P. K.↗
DOE OSTI2021

Anaerobic Dynamic Membrane Bioreactor Development to Facilitate Organic Waste Conversion to Medium-Chain Carboxylic Acids and Their Downstream Recovery

Platform chemicals such as medium-chain carboxylic acids (MCCAs) can be produced from organic waste streams via chain elongation in anaerobic mixed-culture bioreactors. A product recovery system is needed to collect MCCAs from the bioreactor effluent. Membrane-based liquid–liquid extraction, the most commonly used product recovery approach, requires suspended solids removal from the bioreactor effluent to avoid membrane fouling. An anaerobic dynamic membrane bioreactor (AnDMBR) was developed to evaluate MCCA production from brewery and prefermented food waste and to produce a permeate with low suspended solids to facilitate downstream product recovery. The AnDMBR employed an inexpensive stainless-steel mesh as the support material for the development of a biofilm or dynamic membrane, which was responsible for solids–liquid separation. The AnDMBR produced a permeate quality with an average total suspended solids (TSS) concentration of 0.12 g L –1 , while the average bioreactor TSS concentration was two orders of magnitude higher (21.6 g L –1 ). A maximum solids removal efficiency of ≥99% was achieved and good permeate quality was sustained for over 200 days without fouling control or cleaning the support material. In addition to solids–liquid separation, the dynamic membrane was responsible for a substantial fraction of the biological activity of the AnDMBR. The relative activity of Clostridiales, as determined by 16S rRNA sequencing, correlated with MCCA production and was higher in the dynamic membrane (20.0 ± 4.9%) than in the suspended biomass (5.2 ± 2.7%) in the bioreactor. This observation was consistent with MCCA production data as the permeate MCCA concentrations were significantly (p = 8.2 × 10 –5 ) higher than that in the bioreactor, suggesting that the dynamic membrane biofilm contributed to chain elongation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161 – 18 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch, a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes are proving an effective control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161-21 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch and a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes have demonstrated control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161-21 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch and a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes have demonstrated control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRSDate not supplied

CDC Bioreactor Configuration Method for Volume Level Control with Controlled Inlet and Outlet Flow

Environmental Control Life Support Systems and other microbiological biofilm studies often utilize small scale bioreactors. Among these options are the popular BioSurface CDC bioreactors, currently being utilized by Marshall Space Flight Center groups researching the impact of biofilms on life support systems. After a recent experimental regime, it was determined additional equipment could be added to augment the capabilities of the bioreactors. Previous research configurations such as ASTM E3161-21 relied on an outlet stream gravity draining from a side drain port located above sampling coupons. This limited applications to experimental conditions with a controlled inlet and uncontrolled outlet flow. With the introduction of a small single pole single throw (SPST) reed float switch and a peristaltic pump connected to the outlet drain and a chassis controller, the bioreactor is able to maintain a set level. The modification allows additional variables to be tested, including highly adjustable fill and flush cycles, bioreactor volume, draining and filling control. Once the configuration modifications were implemented through the installation of the new equipment, data was collected to ensure the stability of the level measurements. As the level control switch is a float switch, consideration was taken into account for effects of internal stirring speed, along with effects of inlet and outlet flow rate. Data presented in this study will illustrate the stability and effectiveness of the configuration changes in equipment made to the bioreactor. These configuration changes have demonstrated control method to conduct biofilm mitigation techniques for ECLSS hardware research. The controlled level capabilities allow for a constant drip feed flow rate into the bioreactors, a key aspect of the biofilm mitigation testing.

CDC Bioreactor↗
NASA NTRS1995

On-line removal of volatile fatty acids from CELSS anaerobic bioreactor via nanofiltration

The CELSS (controlled ecological life support system) resource recovery system, which is a waste processing system, uses aerobic and anaerobic bioreactors to recover plants nutrients and secondary foods from the inedible biomass. The anaerobic degradation of the inedible biomass by means of culture of rumen bacteria,generates organic compounds such as volatile fatty acids (acetic, propionic, butyric, VFA) and ammonia. The presence of VFA in the bioreactor medium at fairly low concentrations decreases the microbial population's metabolic reactions due to end-product inhibition. Technologies to remove VFA continuously from the bioreactor are of high interest. Several candidate technologies were analyzed, such as organic solvent liquid-liquid extraction, adsorption and/or ion exchange, dialysis, electrodialysis, and pressure driven membrane separation processes. The proposed technique for the on-line removal of VFA from the anaerobic bioreactor was a nanofiltration membrane recycle bioreactor. In order to establish the nanofiltration process performance variables before coupling it to the bioreactor, a series of experiments were carried out using a 10,000 MWCO tubular ceramic membrane module. The variables studied were the bioreactor slurry permeation characteristics, such as, the permeate flux, VFA and the nutrient removal rates as a function of applied transmembrane pressure, fluid recirculation velocity, suspended matter concentration, and process operating time. Results indicate that the permeate flux, VFA and nutrients removal rates are directly proportional to the fluid recirculation velocity in the range between 0.6 to 1.0 m/s, applied pressure when these are low than 1.5 bar, and inversely proportional to the total suspended solids concentration in the range between 23,466 to 34,880. At applied pressure higher than 1.5 bar the flux is not more linearly dependent due to concentration polarization and fouling effects over the membrange surface. It was also found that the permeate flux declines rapidly during the first 5 to 8 hours, and then levels off with a diminishing rate of flux decay.

Colon, Guillermo↗
DOE OSTI2014

New generation NMR bioreactor coupled with high-resolution NMR spectroscopy leads to novel discoveries in Moorella thermoaceticum metabolic profiles

An in-situ nuclear magnetic resonance (NMR) bioreactor was developed and employed to monitor microbial metabolism under batch-growth conditions in real time. We selected Moorella thermoacetica ATCC 49707 as a test case. M. thermoacetica (formerly Clostridium thermoaceticum) is a strictly anaerobic, thermophilic, acetogenic, gram-positive bacterium with potential for industrial production of chemicals. The metabolic profiles of M. thermoacetica were characterized during growth in batch mode on xylose (a component of lignocellulosic biomass) using the new generation NMR bioreactor in combination with high-resolution, high sensitivity NMR (HR-NMR) spectroscopy. In-situ NMR measurements were performed using water-suppressed H-1 NMR spectroscopy at an NMR frequency of 500 MHz, and aliquots of the bioreactor contents were taken for 600 MHz HR-NMR spectroscopy at specific intervals to confirm metabolite identifications and expand metabolite coverage. M. thermoacetica demonstrated the metabolic potential to produce formate, ethanol and methanol from xylose, in addition to its known capability of producing acetic acid. Real-time monitoring of bioreactor conditions showed a temporary pH decrease, with a concomitant increase in formic acid during exponential growth. Fermentation experiments performed outside of the magnet showed that the strong magnetic field employed for NMR detection did not significantly affect cell metabolism. Use of the in-situ NMR bioreactor facilitated monitoring of the fermentation process in real time, enabling identification of intermediate and end-point metabolites and their correlation with pH and biomass produced during culture growth. Real-time monitoring of culture metabolism using the NMR bioreactor in combination with the HR-NMR spectroscopy will allow optimization of the metabolism of microorganisms producing valuable bioproducts.

Bioproducts↗
NASA NTRSDate not supplied

The membrane bioreactor (MBR): A hybrid technology for bioregenerative wastewater treatment and resource recovery in space

Extraplanetary surface habitat life support systems (LSS) on the Moon and Mars, as well as long-duration space travel, will require novel capabilities to withstand anticipated unique, harsh conditions. In order to provide safe, habitable environments for the crew, water purification and waste processing systems will be required to treat all sources of water (condensate, Sabatier, urine, hygiene, fecal, food waste) in order to achieve the necessary levels of recovery needed to sustain life over the long-duration missions. The ability to recycle organic wastes creates an opportunity to recover critical elements (e.g., C, H, O, N, P) for subsequent food production, water purification, and atmospheric regeneration. Bioregenerative systems mimic functions of nature in engineered systems, or bioreactors, utilizing combination of prokaryotes, eukaryotes and archaea. While these systems are commonly used on Earth for wastewater treatment, bioreactors for space travel face additional challenges. Terrestrial bioreactors often rely on gravitational settling of dense flocs and granules for cell retention. For micro- or partial-gravity environments, density differential alone will not be adequate for cell retention; a gravity-independent means for cell retention is crucial. The membrane bioreactor represents the state of the art in wastewater treatment. This hybrid system combines biological processes with membrane filtration to achieve performance beyond what each can accomplish individually. The complete cell retention in an MBR allows for the decoupling of hydraulic retention time (HRT) and solids retention time (SRT), which result in a high-thruput, compact, treatment system. The Bioregenerative Water Technology Team at NASA Kennedy Space Center and the University of South Florida has developed a bioregenerative platform based on the hybrid MBR technology. The overall architecture is compact, modular, flexible, and adaptable to mission evolutions. The main subsystems of the bioregenerative architecture include: 1) Anaerobic membrane bioreactor (AnMBR): Also termed the Organic processor assembly (OPA), the function of the AnMBR is to treat organic wastes such as fecal and food wastes. These wastes are characterized by a concentration of suspended solids comprised of carbohydrates, proteins and lipids. The assigned function of the AnMBR is to break down and covert suspended solids to biogas (methane, hydrogen and carbon dioxide), reduce effluent chemical oxygen demand (COD), liberate organically-bound nutrients, and remove pathogenic organisms. 2) Phototrophic membrane bioreactor (PMBR): The PMBR is comprised of a co-culture of microalgae and bacteria. The assigned function of the PMBR is to polish the permeate of the AnMBR to further remove dissolved organic carbon, manage nutrients (nitrogen transformation, load dampening), and perform air revitalization. 3) Food processor assembly (FPA): The FPA is a food production platform (prokaryotic or eukaryotic), fueled by outputs from the AnMBR, or PMBR. For the presentation, we will describe each step of the bioregenerative architecture, and present performance data from extended trials treating analog and real metabolic wastes.

Bioreactor↗
DOE OSTI2019

Real‐time quantification and supplementation of bioreactor amino acids to prolong culture time and maintain antibody product quality

Abstract Real‐time monitoring of cell cultures in bioreactors can enable expedited responses necessary to correct potential batch failure perturbations which may normally go undiscovered until the completion of the batch and result in failure. Currently, analytical technologies are dedicated to real‐time monitoring of bioreactor parameters such as pH, dissolved oxygen, and temperature, nutrients such as glucose and glutamine, or metabolites such as lactate. Despite the importance of amino acids as the building blocks of therapeutic protein products, other than glutamine their concentrations are not commonly measured. Here, we present a study into amino acid monitoring, supplementation strategies, and how these techniques may impact the cell growth profiles and product quality. We used preliminary bioreactor runs to establish baselines by determining initial amino acid consumption patterns, the results of which were used to select a pool of amino acids which gets depleted in the bioreactor. These amino acids were combined into blends which were supplemented into bioreactors during a subsequent run, the concentrations of which were monitored using a mass spectrometry based at‐line method we developed to quickly assess amino acid concentrations from crude bioreactor media. We found that these blends could prolong culture life, reversing a viable cell density decrease that was leading to batch death. Additionally, we assessed how these strategies might impact protein product quality, such as the glycan profile. The amino acid consumption data were aligned with the final glycan profiles in principal component analysis to identify which amino acids are most closely associated with glycan outcomes.

Powers, David N.↗
DOE OSTI2018

Role of Bioreactors in Microbial Biomass and Energy Conversion

Bioenergy is the world’s largest contributor to the renewable and sustainable energy sector, and it plays a significant role in various energy industries. A large amount of research has contributed to the rapidly evolving field of bioenergy and one of the most important topics is the use of the bioreactor. Bioreactors play a critical role in the successful development of technologies for microbial biomass cultivation and energy conversion. In this chapter, after a brief introduction to bioreactors (basic concepts, configurations, functions, and influencing factors), the applications of the bioreactor in microbial biomass, microbial biofuel conversion, and microbial electrochemical systems are described. Importantly, the role and significance of the bioreactor in the bioenergy process are discussed to provide a better understanding of the use of bioreactors in managing microbial biomass and energy conversion.

Zhang, Liang↗