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At least 91 records · Page 5
All sourcesNASA NTRSDOE OSTI
DOE OSTI2026

Genetic and metabolic drivers of membrane remodeling in Clostridium thermocellum under alcohol stress

Clostridium thermocellum is a leading candidate for consolidated bioprocessing of lignocellulosic biomass into biofuels due to its native cellulolytic capabilities. Beyond ethanol, C. thermocellum is being developed as a platform for producing higher-chain alcohols such as isobutanol and n-butanol. However, its physiological adaptations to alcohol stress remain poorly understood. Here, we investigate how C. thermocellum remodels its membrane lipid composition in response to exogenous ethanol, n-butanol, isobutanol, and butyrate. Exposure to linear alcohols such as n-butanol or to organic acids like butyrate increased the proportion of straight-chain fatty acids in the membrane at the expense of branched-chain species, whereas exposure to the branched alcohol isobutanol produced the opposite effect. Isotope tracer experiments demonstrated that C. thermocellum directly incorporates the carbon backbones of exogenous alcohols and acids into fatty acids, providing a mechanistic basis for these contrasting shifts. We show that the bifunctional aldehyde/alcohol dehydrogenase AdhE is essential for the assimilation of exogenous alcohols into fatty acids, acting through its oxidative activity by first oxidizing alcohols to aldehydes and then converting them to acyl-CoA intermediates. Deletion of the pyruvate:ferredoxin oxidoreductase isozyme pfor4 abolished branched-chain fatty acid synthesis, but supplementation with isobutanol restored production, indicating that Pfor4 substitutes for the canonical branched-chain α-keto acid dehydrogenase complex. These findings reveal two distinct routes for branched-chain fatty acid production in C. thermocellum: a Pfor4-dependent pathway from α-keto acid intermediates derived from amino acid synthesis, and an AdhE-dependent salvage pathway that assimilates exogenous branched-chain alcohols.

Acetivibrio thermocellus↗
DOE OSTI2020

New perspectives on butyrate assimilation in Rhodospirillum rubrum S1H under photoheterotrophic conditions

Background: The great metabolic versatility of the purple non-sulfur bacteria is of particular interest in green technology. Rhodospirillum rubrum S1H is an α-proteobacterium that is capable of photoheterotrophic assimilation of volatile fatty acids (VFAs). Butyrate is one of the most abundant VFAs produced during fermentative biodegradation of crude organic wastes in various applications. While there is a growing understanding of the photoassimilation of acetate, another abundantly produced VFA, the mechanisms involved in the photoheterotrophic metabolism of butyrate remain poorly studied. Results: In this work, we used proteomic and functional genomic analyses to determine potential metabolic pathways involved in the photoassimilation of butyrate. We propose that a fraction of butyrate is converted to acetyl-CoA, a reaction shared with polyhydroxybutyrate metabolism, while the other fraction supplies the ethylmalonyl-CoA (EMC) pathway used as an anaplerotic pathway to replenish the TCA cycle. Surprisingly, we also highlighted a potential assimilation pathway, through isoleucine synthesis and degradation, allowing the conversion of acetyl-CoA to propionyl-CoA. We tentatively named this pathway the methylbutanoyl-CoA pathway (MBC). An increase in isoleucine abundance was observed during the early growth phase under butyrate condition. Nevertheless, while the EMC and MBC pathways appeared to be concomitantly used, a genome-wide mutant fitness assay highlighted the EMC pathway as the only pathway strictly required for the assimilation of butyrate. Conclusion: Photoheterotrophic growth of Rs. rubrum with butyrate as sole carbon source requires a functional EMC pathway. In addition, a new assimilation pathway involving isoleucine synthesis and degradation, named the methylbutanoyl-CoA (MBC) pathway, could also be involved in the assimilation of this volatile fatty acid by Rs. rubrum.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Muconic Acid Production from P. putida Using High Protein Algae Hydrolysate

The composition of algal biomass is highly dynamic, with protein, lipid, and carbohydrate contents varying in response to nutrient and environmental conditions during cultivation. Because shifts in biomass composition are often associated with reduced biomass productivity, production costs can often increase if targeting higher biomass compositional quality (enriched in carbohydrates or lipids at reduced protein content) as input for the algal biorefinery. The optimal algal biorefinery configuration is thus a function of many factors. One of the key strengths of the Combined Algal Processing (CAP) process is the versatility of feedstocks and products produced. The concept has been demonstrated with ethanol and a variety of carboxylic acids (succinic, butyric, muconic) as coproducts along with lipid upgrading to biofuel. Modification of the approaches, processes and downstream upgrading to fuels has allowed the CAP process to reduce costs and improve efficiency. Muconic acid is a high-value, potential fermentation coproduct of interest because it can be easily converted to adipic acid, a high-volume monomer for the production of nylon and other valuable consumer plastics. As such, the production of muconic acid through CAP was explored to expand the suite of products from algal biomass and to begin exploring the valorization of high protein content biomass from rapidly grown algae biomass. We have established initial performance parameters and shown that the range of substrates consumed by the muconic acid-producing microbe, Pseudomonas putida, includes at least glucose, mannose, glycerol, and lactic acid. We achieved complete utilization of these four major hydrolysate substrates achieving productivities of 0.037 (g/L/h) from Scenedesmus obliquus and 0.029 (g/L/h) from Monoraphidium minutum hydrolysates. Final titer and process yield (mol of muconic acid per mol of substrate (molP/molS)) were 0.99 g/L and 0.42 molP/molS from S. obliquus hydrolysate and 0.75 g/L and 0.23 molP/molS from M. minutum hydrolysate, respectively.

BIOMASS FUELS↗
DOE OSTI2020

Development of Clostridium tyrobutyricum as a Microbial Cell Factory for the Production of Fuel and Chemical Intermediates From Lignocellulosic Feedstocks

Microbial conversion of lignocellulosic substrates to fuel and platform chemical intermediates offers a sustainable route to establish a viable bioeconomy. However, such approaches face a series of key technical, economic, and sustainability hurdles, including: incomplete substrate utilization, lignocellulosic hydrolysate, and/or end-product toxicity, inefficient product recovery, incompatible cultivation requirements, and insufficient productivity metrics. Development of a production host with native traits suitable for high productivity conversion of lignocellulosic substrates under process-relevant conditions offers a means to bypass the above-described hurdles and accelerate the development of microbial biocatalyst deployment. Clostridium tyrobutyricum , a native producer of short chain fatty acids, displays a series of characteristics that make it an ideal candidate for conversion of lignocellulosic substrates and thus represents a promising host for microbial production of diverse carboxylate-derived product suites. Herein, recent progress and future directions in the development of this bacterium as an industrial microbial cell factory, with emphases on the utilization of lignocellulosic substrates and metabolic engineering approaches, is reviewed.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Organic Acid Aerosol Measurements from the Mount Airy Site for CoURAGE

This study investigates the prevalence and distribution of organic acid aerosols in a rural environment using filter-based measurements collected in Mount Airy, Maryland, during the CoURAGE campaign from March 19th through June 12th 2025. PM2.5 filters quantify a range of organic acids commonly associated with secondary organic aerosol formation and atmospheric oxidation processes. Each filter was collected using a 15 LPM sampler and was extracted in ultrapure Millipore water (>18 MΩ), allowing water-soluble organic acids to be extracted into solution for analysis. The extracts were then examined using a Waters Acquity I-Class PLUS liquid chromatography system coupled to a Bruker Maxis-II ultra-high-resolution Q-TOF mass spectrometer with electrospray ionization, providing high-sensitivity detection and separation of target compounds. Concentrations of several key organic acids were quantified, including acetic, propionic, pyruvic, butyric, oxalic, isovaleric, valeric, malonic, maleic, succinic, glutaric, malic, adipic, and citric acids. These findings contribute to ongoing efforts to understand regional aerosol composition and their impacts on aerosol-cloud interactions.

Acetic acid↗
DOE OSTI2023

Wetlands harbor lactic acid-driven chain elongators

Wetlands are globally significant carbon storage hotspots. Recent research has suggested that microbially derived metabolites may contribute to soil organic matter formation. Identifying pathways driving the formation of such metabolites is critical to understand the global impact of wetland carbon cycling. Here, we evaluate the presence of chain-elongating organisms converting two to three carbon compounds (i.e., lactic and acetic acid) to medium-chain carboxylic acids (MCCA; i.e., six-carbon caproic acid) in wetland soils. We demonstrate the enrichment of a lactic acid-driven chain-elongating community from wetland soils producing a mixture of butyric and caproic acid. The enriched community was dominated by Clostridiaceae, Ruminococcaceae, and Lachnospiraceae, three families with known chain elongators. Amplicon sequencing identified three Ruminococcaceae and one Clostridiaceae zero-radius OTU (zOTU) that were (i) present in the soil, (ii) enriched over 1% relative abundance in the bioreactor, and (iii) were closely related to known chain elongators. Moreover, close relatives of the three Ruminococcaceae zOTU were also observed in several other wetland microbiomes. From this observation, we conclude that close relatives of known chain elongators, potentially capable of lactic acid-driven MCCA production themselves, are present in wetland soils. This observation may have implications for our understanding of carbon cycling and storage in wetland ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2002

Fish oil increases mitochondrial phospholipid unsaturation, upregulating reactive oxygen species and apoptosis in rat colonocytes

We have shown that a combination of fish oil (high in n-3 fatty acids) with the butyrate-producing fiber pectin, upregulates apoptosis in colon cells exposed to the carcinogen azoxymethane, protecting against colon tumor development. We now hypothesize that n-3 fatty acids prime the colonocytes such that butyrate can initiate apoptosis. To test this, 30 Sprague-Dawley rats were provided with diets differing in the fatty acid composition (corn oil, fish oil or a purified fatty acid ethyl ester diet). Intact colon crypts were exposed ex vivo to butyrate, and analyzed for reactive oxygen species (ROS), mitochondrial membrane potential (MMP), translocation of cytochrome C to the cytosol, and caspase-3 activity (early events in apoptosis). The fatty acid composition of the three major mitochondrial phospholipids was also determined, and an unsaturation index calculated. The unsaturation index in cardiolipin was correlated with ROS levels (R = 0.99; P = 0.02). When colon crypts from fish oil and FAEE-fed rats were exposed to butyrate, MMP decreased (P = 0.041); and translocation of cytochrome C to the cytosol (P = 0.037) and caspase-3 activation increased (P = 0.032). The data suggest that fish oil may prime the colonocytes for butyrate-induced apoptosis by enhancing the unsaturation of mitochondrial phospholipids, especially cardiolipin, resulting in an increase in ROS and initiating apoptotic cascade.

Non-NASA Center↗
DOE OSTI2024

Adaptive laboratory evolution and metabolic engineering of Cupriavidus necator for improved catabolism of volatile fatty acids

Bioconversion of high-volume waste streams into value-added products will be an integral component of the growing bioeconomy. Volatile fatty acids (VFAs) (e.g., butyrate, valerate, and hexanoate) are an emerging and promising waste-derived feedstock for microbial carbon upcycling. Cupriavidus necator H16 is a favorable host for conversion of VFAs into various bioproducts due to its diverse carbon metabolism, ease of metabolic engineering, and use at industrial scales. Here, in this study, we report that a common strategy to improve product titers in C. necator, deletion of the polyhydroxybutyrate (PHB) biosynthetic operon, results in a significant growth defect on VFA substrates. Using adaptive laboratory evolution, we identify mutations to the regulator gene phaR, the two-component response regulator-histidine kinase pair encoded by H16_A1372/H16_A1373, and the tripartite transporter assembly encoded by H16_A2296-A2298 as causative for improved growth on VFA substrates. Deletion of phaR and H16_A1373 led to significantly reduced NADH abundance accompanied by large changes to expression of genes involved in carbon metabolism, balance of electron carriers, and oxidative stress tolerance that may be responsible for improved growth of these engineered strains. These results provide insight into the role of PHB biosynthesis in carbon and energy metabolism and highlight a key role for the regulator PhaR in global regulatory networks. By combining mutations, we generated platform strains with significant growth improvements on VFAs, which can enable improved conversion of waste-derived VFA substrates to target bioproducts.

09 BIOMASS FUELS↗
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 OSTI2023

A metagenome-level analysis of a microbial community fermenting ultra-filtered milk permeate

Fermentative microbial communities have the potential to serve as biocatalysts for the conversion of low-value dairy coproducts into renewable chemicals, contributing to a more sustainable global economy. To develop predictive tools for the design and operation of industrially relevant strategies that utilize fermentative microbial communities, there is a need to determine the genomic features of community members that are characteristic to the accumulation of different products. To address this knowledge gap, we performed a 282-day bioreactor experiment with a microbial community that was fed ultra-filtered milk permeate, a low-value coproduct from the dairy industry. The bioreactor was inoculated with a microbial community from an acid-phase digester. A metagenomic analysis was used to assess microbial community dynamics, construct metagenome-assembled genomes (MAGs), and evaluate the potential for lactose utilization and fermentation product synthesis of community members represented by the assembled MAGs. This analysis led us to propose that, in this reactor, members of the Actinobacteriota phylum are important in the degradation of lactose, via the Leloir pathway and the bifid shunt, and the production of acetic, lactic, and succinic acids. In addition, members of the Firmicutes phylum contribute to the chain-elongation-mediated production of butyric, hexanoic, and octanoic acids, with different microbes using either lactose, ethanol, or lactic acid as the growth substrate. We conclude that genes encoding carbohydrate utilization pathways, and genes encoding lactic acid transport into the cell, electron confurcating lactate dehydrogenase, and its associated electron transfer flavoproteins, are genomic features whose presence in Firmicutes needs to be established to infer the growth substrate used for chain elongation.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Genomic and functional analyses of fungal and bacterial consortia that enable lignocellulose breakdown in goat gut microbiomes

Abstract The herbivore digestive tract is home to a complex community of anaerobic microbes that work together to break down lignocellulose. These microbiota are an untapped resource of strains, pathways and enzymes that could be applied to convert plant waste into sugar substrates for green biotechnology. We carried out more than 400 parallel enrichment experiments from goat faeces to determine how substrate and antibiotic selection influence membership, activity, stability and chemical productivity of herbivore gut communities. We assembled 719 high-quality metagenome-assembled genomes (MAGs) that are unique at the species level. More than 90% of these MAGs are from previously unidentified herbivore gut microorganisms. Microbial consortia dominated by anaerobic fungi outperformed bacterially dominated consortia in terms of both methane production and extent of cellulose degradation, which indicates that fungi have an important role in methane release. Metabolic pathway reconstructions from MAGs of 737 bacteria, archaea and fungi suggest that cross-domain partnerships between fungi and methanogens enabled production of acetate, formate and methane, whereas bacterially dominated consortia mainly produced short-chain fatty acids, including propionate and butyrate. Analyses of carbohydrate-active enzyme domains present in each anaerobic consortium suggest that anaerobic bacteria and fungi employ mostly complementary hydrolytic strategies. The division of labour among herbivore anaerobes to degrade plant biomass could be harnessed for industrial bioprocessing.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1999

Syntrophus aciditrophicus sp. nov., a new anaerobic bacterium that degrades fatty acids and benzoate in syntrophic association with hydrogen-using microorganisms

Strain SBT is a new, strictly anaerobic, gram-negative, nonmotile, non-sporeforming, rod-shaped bacterium that degrades benzoate and certain fatty acids in syntrophic association with hydrogen/formate-using microorganisms. Strain SBT produced approximately 3 mol of acetate and 0.6 mol of methane per mol of benzoate in coculture with Methanospirillum hungatei strain JF1. Saturated fatty acids, some unsaturated fatty acids, and methyl esters of butyrate and hexanoate also supported growth of strain SBT in coculture with Desulfovibrio strain G11. Strain SBT grew in pure culture with crotonate, producing acetate, butyrate, caproate, and hydrogen. The molar growth yield was 17 +/- 1 g cell dry mass per mol of crotonate. Strain SBT did not grow with fumarate, iron(III), polysulfide, or oxyanions of sulfur or nitrogen as electron acceptors with benzoate as the electron donor. The DNA base composition of strain SBT was 43.1 mol% G+C. Analysis of the 16 S rRNA gene sequence placed strain SBT in the delta-subdivision of the Proteobacteria, with sulfate-reducing bacteria. Strain SBT was most closely related to members of the genus Syntrophus. The clear phenotypic and genotypic differences between strain SBT and the two described species in the genus Syntrophus justify the formation of a new species, Syntrophus aciditrophicus.

Non-NASA Center↗
NASA NTRS1996

Anaerobic degradation of inedible crop residues produced in a Controlled Ecological Life Support System

An anaerobic reactor seeded with organisms from an anaerobic lagoon was used to study the degradation of inedible crop residues from potato and wheat crops grown in a closed environment. Conversion of this biomass into other products was also evaluated. Degradation of wheat volatile solids was about 25% where that of potato was about 50%. The main product of the anaerobic fermentation of both crops was acetic acid with smaller quantities of propionate and butyrate produced. Nitrate, known to be high in concentration in inedible potato and wheat biomass grown hydroponically, was converted to ammonia in the anaerobic reactor. Both volatile fatty acid and ammonia production may have implications in a crop production system.

NASA Program Advanced Life Support↗
NASA NTRS1977

Formation of fatty acids in photochemical conversions of saturated hydrocarbons

Abiogenic synthesis of fatty acids was studied in photochemical conversions of saturated hydrocarbons. It was shown that, in a hydrocarbon water CaCO3 suspension, the action of 254 nm UV rays caused the formation of fatty acids with a maximum number of carbon atoms in the chain not exceeding that in the initial hydrocarbon. Synthesis of acetic, propionic, butyric, valeric, caproic, enanthic and caprylic (in the case of octane) acids occurs in heptane water CaCO3 and octane water CaCO3 systems.

Telegina, T. A.↗
DOE OSTI2022

The effects of xylo-oligosaccharides on regulating growth performance, nutrient utilization, gene expression of tight junctions, nutrient transporters, and cecal short chain fatty acids profile in Eimeria-challenged broiler chickens

A 21-d experiment was conducted to investigate the effects of xylo-oligosaccharides (XOS) on growth performance, nutrient utilization, gene expression of tight junctions, nutrient transporters, and cecal short chain fatty acids (SCFA) profile of broiler chickens challenged with mixed Eimeria spp. Two hundred fifty-two zero-day-old chicks were allocated to 6 treatments in a 3 £ 2 factorial arrangement (corn-soybean meal diets supplemented with 0, 0.5, or 1.0 g/kg XOS; with or without Eimeria challenge). Challenged groups were inoculated with a solution containing E. maxima, E. acervulina, and E. tenella oocysts on d 15. During the infection period (d 15 to d 21), there was a significant (P < 0.05) Eimeria £ XOS interaction for weight gain (WG). XOS significantly (P < 0.05) increased WG in the unchallenged birds but not in the challenged treatments. There was no significant Eimeria £ XOS interaction for N and minerals utilization responses. XOS supplementation at 0.5 g/kg tended to alleviate Eimeria-induced depression in apparent ileal digestibility of DM (P = 0.052). Challenged birds had lower (P < 0.01) AME, AMEn, and total retention of N, Ca, and P. Eimeria upregulated (P < 0.01) gene expression of tight junction proteins claudin-1, junctional adhesion molecule-2, and glucose transporter GLUT1; but downregulated (P < 0.01) the peptide transporter PepT1, amino acid transporters rBAT, CAT2, y+LAT2, and zinc transporter ZnT1. XOS alleviated (P < 0.05) Eimeria-induced claudin-1 upregulation. Eimeria decreased (P < 0.05) cecal saccharolytic SCFA acetate, butyrate, and total SCFA, but increased (P < 0.05) branched chain fatty acids isobutyrate and isovalerate. The supplementation of XOS tended to decrease the concentration of isobutyrate (P = 0.08) and isovalerate (P = 0.062). In conclusion, 0.5 g/kg XOS supplementation alleviated depression in growth performance and nutrient utilization from the Eimeria challenge. In addition, supplemental XOS reversed the gene expression changes of claudin-1, also showed the potentials of alleviating the negative cecal fermentation pattern induced by Eimeria infection.

60 APPLIED LIFE SCIENCES↗