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At least 19 records
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Optimization of Spirulina for biomanufacturing and the delivery of protein therapeutics

Lumen Bioscience has developed a novel cyanobacterial platform that enormously decreases the cost of pharmaceuticals used to prevent and treat illnesses. Arthrospira platensis (spirulina), is a photosynthetic microorganism that has been consumed as a dietary supplement for centuries in part because of its high protein content. Commercial cultivation operations have matured over the last 50 years to allow large-scale cultivation in outdoor ponds. Lumen recently discovered methods that transform this commercially important cyanobacterium into a genetically tractable platform for bioengineering. The high protein accumulation in spirulina makes it an ideal chassis for heterologous protein expression, and centuries of safe human consumption suggest its utility as an administration vehicle for biologic drugs. Lumen can express a broad range of recombinant proteins in spirulina and has developed manufacturing processes and strains to mucosally deliver bioactive proteins to treat and/or prevent disease. For example, Lumen has produced an oral cocktail of dried whole-cell spirulina biomass, containing 3 toxin-neutralizing antibodies and 1 endolysin, that is efficacious in a preclinical in vivo models of C. difficile infection. Lumen has also demonstrated that intranasal administration of a spirulina-manufactured neutralizing antibody can prevent disease by a respiratory pathogen, SARS-CoV-2, in a hamster model. Lumen’s current pipeline includes therapeutics to treat or prevent C. difficile infection, COVID-19, inflammatory bowel disease, cardiometabolic disease, and traveler’s diarrhea. Lumen deploys strategies to maximize the expression of these therapeutics for optimal dose sizing, a key limiting factor with past food crop-based expression systems. Spirulina can express exogenous proteins at unusually high levels (>15% of dry weight) but reaching this maximum value requires optimization of multiple interacting factors. We describe our current statistics design of experiments approach that optimizes these factors with minimal replicates. This maximizes exogenous protein expression allowing Lumen to generate spirulina-based therapeutics for a rapidly expanding range of medical uses.

Heinnickel, Mark↗
DOE OSTI2022

Development of spirulina for the manufacture and oral delivery of protein therapeutics

The use of the edible photosynthetic cyanobacterium Arthrospira platensis (spirulina) as a biomanufacturing platform has been limited by a lack of genetic tools. Here we report genetic engineering methods for stable, high-level expression of bioactive proteins in spirulina, including large-scale, indoor cultivation and downstream processing methods. Following targeted integration of exogenous genes into the spirulina chromosome (chr), encoded protein biopharmaceuticals can represent as much as 15% of total biomass, require no purification before oral delivery and are stable without refrigeration and protected during gastric transit when encapsulated within dry spirulina. Oral delivery of a spirulina-expressed antibody targeting campylobacter—a major cause of infant mortality in the developing world—prevents disease in mice, and a phase 1 clinical trial demonstrated safety for human administration. Spirulina provides an advantageous system for the manufacture of orally delivered therapeutic proteins by combining the safety of a food-based production host with the accessible genetic manipulation and high productivity of microbial platforms.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Evaluation of Apparent Metabolizable Energy and Apparent Ileal Amino Acid Digestibility of Spirulina (Arthrospira platensis) in Broiler Chickens and Laying Hens

Our study objective was to determine the apparent metabolizable energy (AME) and apparent ileal amino acid digestibility (AIAAD) of spirulina fed to broiler chickens and laying hens using the difference method. In both experiments, birds were either fed corn-soybean meal basal diets, containing no spirulina and formulated to provide the nutrient requirements of either broilers or layers, or fed test diets consisting of 25% spirulina and 75% of the appropriate basal diet. Titanium dioxide was added to all diets as an indigestible marker. The diets were fed to 10 replicate cages/treatment of broilers and 15 replicate cages/treatment of laying hens. The AME of spirulina for broilers was significantly lower (p < 0.05) (2368 ± 104 kcal/kg, as fed) than for laying hens (3144 ± 173 kcal/kg, as fed), suggesting bird type and age may influence energy utilization. The AIAAD of spirulina fed to broiler chickens did not differ from that of laying hens (p > 0.05), except for valine, alanine, and glycine, which were all significantly higher in laying hens (p < 0.05). Overall, the AIAAD for nonessential amino acids averaged 81.1%, with no significant difference between essential and nonessential amino acids. Differences in spirulina nutrient content cited in the literature support further research to determine the optimal inclusion of this alternative ingredient in broiler and layer diets.

Agriculture↗
NASA NTRS1990

Algae for controlled ecological life support system diet characterization of cyanobacteria 'spirulina' in batch cultures

Spirulina sp. is a bioregenerative photosynthetic and edible alga for space craft crews in a Closed Ecological Life Support System (CLESS). It was characterized for growth rate and biomass yield in batch cultures, under various environmental conditions. The cell characteristics were identified for one strain of Spirulina: S. maxima. Fast growth rate and high yield were obtained. The partitioning of the assimulatory products (proteins, carbohydrates, lipids) were manipulated by varying the environmental conditions. Experiments with Spirulina demonstrated that under stress conditions carbohydrate increased at the expense of protein. In other experiments, where the growth media were sufficient in nutrients and incubated under optimum growth conditions, the total proteins were increased up to almost 70 percent of the organic weight. In other words, the nutritional quality of the alga could be manipulated by growth conditions. These results support the feasibility of considering Spirulina as a subsystem in CELSS because of the ease with which its nutrient content can be manipulated.

Tadros, M. G.↗
NASA NTRS1988

Characterization of Spirulina biomass for CELSS diet potential

Spirulina sp. as a bioregenerative photosynthetic and an edible alga for space craft crew in a CELSS, was characterized for growth rate and biomass yield in batch cultures, under various environmental conditions. The cell characteristics were identified for two strains of Spirulina: S. maxima and S. plantensis. Fast growth rate and high yield of both strains were obtained under the following conditions: temperature (30 to 35 C), light irradiance (60 to 100 uE/m/s), nitrate (30 mM), phosphate (2 mM), aeration (300 ml/min), and ph (9 to 10). The partitioning of the assimalatory products (proteins, carbohydrates, lipids) were manipulated by varying the environmental growth conditions. The experiments with Spirulina demonstrated that under stress conditions (high light 120 uE/m/s, temperature 38 C, nitrogen or phosphate limitation; 0.1 M sodium chloride) carbohydrate increased at the expense of protein. In other experiments, where the growth media were sufficient in nutrients and incubated under optimum growth conditions, the total proteins were increased up to almost 70 percent of the organic weight. Conclusion: The nutritional quality of the alga could be manipulated by growth conditions, and therefore usful as a subsystem in CELSS.

Tadros, Mahasin G.↗
NASA NTRS1989

Manipulating cyanobacteria: Spirulina for potential CELSS diet

Spirulina sp. as a bioregenerative photosynthetic and an edible alga for spacecraft crew in a CELSS, was characterized for the biomass yield in batch cultures, under various environmental conditions. The partitioning of the assimalitory products (proteins, carbohydrates, lipids) were manipulated by varying the environmental growth conditions. Experiments with Spirulina have shown that under stress conditions (i.e., high light 160 uE/sq m/s, temperature 38 C, nitrogen or phosphate limitation; 0.1 M sodium chloride) carbohydrates increased at the expense of proteins. In other experiments, where the growth media were sufficient in nutrients and incubated under optimum growth conditions, the total of the algal could be manipulated by growth conditions. These results support the feasibility of considering Spirulina as a subsystem in CELSS because of the ease with which its nutrient content can be manipulated.

Tadros, Mahasin G.↗
NASA NTRS1987

Food production and gas exchange system using blue-green alga (spirulina) for CELSS

In order to reduce the cultivation area required for the growth of higher plants in space adoption of algae, which have a higher photosynthetic ability, seems very suitable for obtaining oxygen and food as a useful source of high quality protein. The preliminary cultivation experiment for determining optimum cultivation conditions and for obtaining the critical design parameters of the cultivator itself was conducted. Spirulina was cultivated in the 6 liter medium containing a sodium hydrogen carbonate solution and a cultivation temperature controlled using a thermostat. Generated oxygen gas was separated using a polypropyrene porous hollow fiber membrane module. Through this experiment, oxygen gas (at a concentration of more than 46 percent) at a rate of 100 to approx. 150 ml per minute could be obtained.

Oguchi, Mitsuo↗
NASA NTRS1993

Characterization of Spirulina biomass for CELSS diet potential

Cyanobacteria, Spirulina maxima as a biogenerative photosynthetic and an edible alga for the space craft crew in a CELSS, was evaluated in an effort to increase the growth rate, biomass, yield, and chemical analysis in continuous cultures. The cell characteristics were determined for cultures maintained at steady state with respect to the substrate concentration. The productivity increased in experiments exposed to low light (30 uE m(exp -2)s(exp -1). Oxygen evolved and protein production were higher in cultures exposed to low light intensity. There was a relationship between nitrate concentration and the yield of the culture. Increasing the concentration of nitrate in the growth medium up to 20 mM was enough to produce a culture having the same chemical composition as that of complete medium. High light was inhibiting the yield of the culture. Increasing the concentration of phosphate beyond 1 mM did not improve the yield of the culture. Increasing the concentration of sodium chloride in the growth medium did not affect the growth of the alga up to 0.1 M but beyond that the culture started to be stressed. The response to stress appeared in high production of total carbohydrate on the expense of protein production. The oxygen production was also higher in cultures stressed with sodium chloride.

Tadros, Mahasin G.↗
NASA NTRS2006

Novel Concept for LSS Based on Advanced Microalgal Biotechnologies

One of the key issues for successful human space exploration is biomedical life support in hostile space and planetary environments that otherwise cannot sustain life. Bioregenerative life support systems (LSS) are one of the options for atmospheric regeneration. To date, no bioregenerative LSS has shown capability for 100% air regeneration. Nor have these LSS been robust enough to simultaneously provide a regenerable complete food source. In contrast to microalgae, traditional plant approaches, e.g. wheat and lettuce, are lacking essential amino acids, vitamins, and micronutrients. Moreover, the rate of photosynthesis by microalgae significantly exceeds that of high plants. Nevertheless, the employment of microalgae in LSS technology was restricted, until recently, due to high water demands. Also the per person requirement of a 40L volume of microalgae in a photobioreactor, to provide daily O2 production, made an algae-based approach less attractive. By employing a vertically stacked membrane bioreactor, coupled with a solar tracker and photon-delivery system, a lightweight air revitalization system for space based applications, with minimal water requirements, can be developed. Our preliminary estimations suggest that a membrane bioreactor, 8m3 in volume, comprised of 80m2 (twenty 2m x 2m membranes, each spaced 10 cm apart), and a total of 70L of water could produce 2.7 kg of dried microalgal biomass that would supply the energy and essential amino acid requirements, as well as producing sufficient O2 for the daily needs of a 15 member crew. Research on the biochemical content of edible blue-green alga Spirulina (Arthrospira) platensis shows a wide spectrum of stable Spirulina mutants with an enhanced content of amino acids, -carotene, and phycobilliprotein c-phycocyanin. Feeding animals suffering from radiation-induced lesions, c-phycocyanin, extracted from strain 27G, led to a correction in the decrement of dehydrogenase activity and energy-rich phosphate levels, as well as improved antioxidant defense and pyruvate levels, compared to untreated animals. Experimental anemia in rats was corrected by feeding Spirulina platensis strains 198B and 27G, (with an enhanced content of methionine, phycobiliproteins and carotenoids). Spirulina was recently shown by Ananyev et al, 2005, to be an oxygenic organism with the highest level of photosystem II activity (O2 production). We propose therefore to develop a design for membrane-based photoreactors for Lunar and Mars exploration habitat LSS, for the cultivation of genetically modified strains of Spirulina to scrub CO2 and supply astronauts with O2, protein, vitamins, and immunostimulators.

Brown, I.↗
NASA NTRS2024

Space Algae-2 Ground and Lunar Analog Studies in Preparation for Long-Duration Propagation of Cyanobacteria in Spaceflight

There are numerous applications for microalgae in spaceflight missions and on Earth, such as oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments optimized conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and the protein and metabolite profile. The concept of operations for Space Algae-2 was tested during a lunar mission simulation within a semi-controlled environment. During a six-day lunar analog mission at the Hawai’i Space Exploration Analog and Simulation (HI-SEAS) A. platensis was successfully grown using flight-like hardware. The cyanobacteria were harvested and used to supplement bread as an example of spirulina biomass utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.

Algae↗
NASA NTRSDate not supplied

Space Algae-2 Ground and Lunar Analog Studies in Preparation for Long-Duration Propagation of Cyanobacteria in Spaceflight

There are numerous applications for microalgae in spaceflight missions and on Earth, such as oxygen production, carbon dioxide removal, nutrition, wastewater processing, and biofuel production. Space Algae-2 aims to test the genetic stability of Arthrospira platensis, commonly known as spirulina, during six-months of continuous culture on the International Space Station. Long-duration exposure to ionizing radiation and microgravity may impact growth, nutrient composition, and genetic stability. The high protein, vitamin, antioxidant content, and radiation resistance make spirulina a promising candidate for bioregenerative life support systems. A concept of operations was developed to grow and harvest algal biomass in space. Preflight testing experiments optimized conditions for an extended growth period in a gas permeable bioreactor bag. Preflight and post-harvest storage methods were developed in addition to a novel cryopreservation method. After sample return, multi-omics analysis will be conducted to determine the mutation rate, gene expression, and the protein and metabolite profile. The concept of operations for Space Algae-2 was tested during a lunar mission simulation within a semi-controlled environment. During a six-day lunar analog mission at the Hawai’i Space Exploration Analog and Simulation (HI-SEAS) A. platensis was successfully grown using flight-like hardware. The cyanobacteria were harvested and used to supplement bread as an example of spirulina biomass utilization. Overall, the data collected from Space Algae-2 will inform potential bioengineering of spirulina for space and terrestrial applications.

Algae↗
NASA NTRS1987

Fundamental study on gas monitoring in CELSS

A mass spectrometer and computer system was developed for conducting a fundamental study on gas monitoring in a Controlled Ecological Life Support System. Respiration and metabolism of the hamster and photosynthesis of the Spirulina were measured in a combination system consisting of a hamster chamber and a Spirulina cultivator. They are connected through a membrane gas exchanger. Some technical problems were examined. In the mass spectrometric gas monitoring, a simultaneous multisample measurement was developed by employing a rotating exchange valve. Long term precise measurement was obtained by employing an automatic calibration system. The membrane gas sampling probe proved to be useful for long term measurement. The cultivation rate of the Spirulina was effectively changed by controlling CO2 and light supply. The experimental results are helpful for improving the hamster-spirulina system.

Nishi, I.↗
NASA NTRSDate not supplied

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗
NASA NTRSDate not supplied

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗