SEARCH · Engineering Papers

Results for “Product”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 505 records · Page 28

All sources NASA NTRS DOE OSTI

DOE OSTI2020

Numerical study of the effect of propped surface area and fracture conductivity on shale gas production: Application for multi-size proppant pumping schedule design

Hydraulic fracturing is a technique extensively used in the oil and gas industry, where water, proppant (sand) and additives are injected into unconventional reservoirs to enhance the recovery of shale hydrocarbon. Although some previous studies have developed pumping schedules that maximize gas production for a single-size proppant, there are very few studies that consider the effect of varying proppant diameters across pumping stages on shale gas production. Motivated by this, we carried out an extensive sensitivity analysis to determine the effect of different proppant diameters on the average fracture conductivity (FC), average propped surface area (PSA) and cumulative shale gas production volume. We found out that the cumulative shale gas production volume depends on both the average PSA and average FC. We also found out that small-diameter proppant resulted in higher average PSA and lower average FC, whereas large-diameter proppant resulted in lower average PSA and higher average FC. Hence, we designed a multi-size proppant pumping schedule considering both of these parameters into account for simultaneously propagating multiple fractures to maximize shale gas production from unconventional reservoirs. Since the size of injected proppant particles determines the average PSA and average FC for the propped hydraulic fractures, we developed a novel framework called Sequentially Interlinked Modeling Structure (SIMS) to predict the average PSA, average FC and cumulative shale gas production volume at the end of 10 years for a given pumping schedule. Then, we used this SIMS framework to obtain a multi-size proppant pumping schedule that maximizes shale gas production. In closing, we demonstrated that the obtained pumping schedule gives a cumulative shale gas production volume greater than the values obtained from the existing pumping schedules.

03 NATURAL GAS↗

DOE OSTI2024

Measurement of 𝐽∕𝜓and 𝜓(2𝑆)production in 𝑝 +𝑝and 𝑝 +𝑑interactions at 120 GeV

We report the p + p and p + d differential cross sections measured in the SeaQuest experiment for J/psi and psi(2S) production at 120GeV beam energy covering the forward x-Feynman (x(F)) range of 0.5 < x(F) < 0.9. The measured cross sections are in good agreement with theoretical calculations based on the nonrelativistic QCD (NRQCD) using the long-distance matrix elements deduced from a recent global analysis of proton- and pion-induced charmonium production data. The sigma(psi(2S))/sigma(J/psi) cross section ratios are found to increase as x(F) increases, indicating that the q (q) over bar. annihilation process has larger contributions in the psi(2S) production than the J/psi production. The sigma(pd)/2 sigma(pp) cross section ratios are observed to be significantly different for the DrellYan process and J/psi production, reflecting their different production mechanisms. We find that the sigma(pd)/2 sigma(pp) ratios for J/psi production at the forward x(F) region are sensitive to the (d) over bar/(u) over bar flavor asymmetry of the proton sea, analogous to the Drell-Yan process. The transverse momentum (p(T)) distributions for J/psi and psi(2S) production are also presented and compared with data collected at higher center-of-mass energies.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗

DOE OSTI2023

Evaluating Multimodel Ensemble Seasonal Climate Forecasts on Rangeland Plant Production in the California Annual Grassland

Seasonal precipitation and temperature directly affect total plant production in the California Annual Grassland (CAG). Technological advances have resulted in skillful seasonal climate forecasts (i.e., significant correlations between actual and forecasted climate), which could be input into plant production models to inform stocking and other rangeland management decisions. This study presents a procedure for forecasting plant production in the CAG ecosystem to predict annual plant production for grazing, restoration, or other rangeland management practices using a combination of historical gridMET climate data and seasonal hindcasts (i.e., retrospective forecasts, from the North American Multi-Model Ensemble program). Further, the results of this study first confirmed high forecast skill, throughout the growing season at all sites. We also identified skillful plant production forecasts across most of the growing season at two sites and in three of the seven forecasting months at one study site. Forecasting climate and end-of-year plant production across the growing season at three CAG sites allowed us to identify the places and times in the growing season when forecasting might be most helpful in informing management decisions. Integrating plant production forecasting into rangeland management practices could significantly improve rangeland management outcomes. These procedures provide a user guide for creating plant production forecasts for any given area of interest and may be applicable across a wide range of other agricultural and rangeland management systems.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Managing weather- and market price-related financial risks in algal biofuel production

Large-scale algae production has garnered interest due to its potential as a biofuel feedstock. Previous research assessing the profitability of algae products has been mostly based on values averaged over time, but algae production and resulting financial returns exhibit significant variability due to weather and fluctuations in selling prices for algae-based products. In other sectors, producers often reduce weather- and market price-related financial risk with financial instruments such as insurance, but little research has been performed on the design of insurance products to protect algae producers. Furthermore, this study develops a novel index-based insurance instrument that pays-out during unfavorable weather and market conditions, then explores the instrument's effectiveness, combined with a cash reserve, in reducing revenue stream variability for an algae producer. Results indicate that a biophysically based index-insurance product tailored to the specific financial risks in algae production can reduce variability in net revenues and can do so at a lower cost than relying solely on cash reserves, the most common financial risk management tool. Assessing the performance of index-insurance in algae production is particularly timely given the passage of the 2018 Farm Bill, which newly opens opportunities for the USDA to provide crop insurance to algae producers.

09 BIOMASS FUELS↗

DOE OSTI2022

Decoupling of greenness and gross primary productivity as aridity decreases

Ecosystem primary productivity is a key ecological process influencing many ecosystem services, including carbon storage. Thus, clarifying how primary productivity in terrestrial ecosystems responds to climatic variability can reveal key mechanisms that will drive future changes in the global carbon budget. Satellite products of canopy greenness are widely used as proxies for vegetation productivity to evaluate how ecosystems respond to climate variability. However, to what degree inter-annual variations in productivity are consistent with greenness and how this relationship varies spatially remains unclear. Here we investigated the strength of the coupling between inter-annual variations in leaf area index (LAI, a measure of greenness) and ecosystem gross primary productivity (GPP) derived from eddy covariance towers, i.e., the r2 of the LAI-GPP relationship. Overall, inter-annual GPP and LAI were highly coupled (i.e., high r2) in arid grasslands, but were fully decoupled in mesic evergreen broadleaf forests, indicating that this relationship varies strongly along aridity gradients. A possible mechanism of the spatial variation in the LAI-GPP relationship is that the tradeoff between ecosystem structure (LAI) and physiology (photosynthesis per unit leaf area) becomes stronger in more humid climates. Land models overestimated the r2 of LAI-GPP correlation for most ecosystem types and failed to capture the spatial pattern along aridity gradients. We conclude that relying on greenness products for evaluating inter-annual changes in vegetation productivity may bias assessments, especially in tropical rainforest ecosystems. Our findings may also reconcile observed disparities between responses in greenness and GPP during drought in Amazon forests.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2023

Fluctuating pH for efficient photomixotrophic succinate production

Cyanobacteria are attracting increasing attention as a photosynthetic chassis organism for diverse biochemical production, however, photoautotrophic production remains inefficient. Photomixotrophy, a method where sugar is used to supplement baseline autotrophic metabolism in photosynthetic hosts, is becoming increasingly popular for enhancing sustainable bioproduction with multiple input energy streams. In this study, the commercially relevant diacid, succinate, was produced photomixotrophically. Succinate is an important industrial chemical that can be used for the production of a wide array of products, from pharmaceuticals to biopolymers. In this system, the substrate, glucose, is transported by a proton symporter and the product, succinate, is hypothesized to be transported by another proton symporter, but in the opposite direction. Thus, low pH is required for the import of glucose and high pH is required for the export of succinate. Succinate production was initiated in a pH 7 medium containing bicarbonate. Glucose was efficiently imported at around neutral pH. Utilization of bicarbonate by CO 2 fixation raised the pH of the medium. As succinate, a diacid, was produced, the pH of the medium dropped. By repeating this cycle with additional pH adjustment, those contradictory requirements for transport were overcome. pH affects a variety of biological factors and by cycling from high pH to neutral pH processes such as CO 2 fixation rates and CO 2 solubility can vary. In this study the engineered strains produced succinate during fluctuating pH conditions, achieving a titer of 5.0gL -1 after 10 days under shake flask conditions. Finally, these results demonstrate the potential for photomixotrophic production as a viable option for the large-scale production of succinate.

36 MATERIALS SCIENCE↗

DOE OSTI2020

Isobutanol production freed from biological limits using synthetic biochemistry

Cost competitive conversion of biomass-derived sugars into biofuel will require high yields, high volumetric productivities and high titers. Suitable production parameters are hard to achieve in cell-based systems because of the need to maintain life processes. As a result, next-generation biofuel production in engineered microbes has yet to match the stringent cost targets set by petroleum fuels. Removing the constraints imposed by having to maintain cell viability might facilitate improved production metrics. Here, we report a cell-free system in a bioreactor with continuous product removal that produces isobutanol from glucose at a maximum productivity of 4 g L –1 h –1 , a titer of 275 g L –1 and 95% yield over the course of nearly 5 days. These production metrics exceed even the highly developed ethanol fermentation process. Our results suggest that moving beyond cells has the potential to expand what is possible for bio-based chemical production.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2023

Utilization of lignocellulosic hydrolysates for photomixotrophic chemical production in Synechococcus elongatus PCC 7942

Abstract To meet the need for environmentally friendly commodity chemicals, feedstocks for biological chemical production must be diversified. Lignocellulosic biomass are an carbon source with the potential for effective use in a large scale and cost-effective production systems. Although the use of lignocellulosic biomass lysates for heterotrophic chemical production has been advancing, there are challenges to overcome. Here we aim to investigate the obligate photoautotroph cyanobacterium Synechococcus elongatus PCC 7942 as a chassis organism for lignocellulosic chemical production. When modified to import monosaccharides, this cyanobacterium is an excellent candidate for lysates-based chemical production as it grows well at high lysate concentrations and can fix CO 2 to enhance carbon efficiency. This study is an important step forward in enabling the simultaneous use of two sugars as well as lignocellulosic lysate. Incremental genetic modifications enable catabolism of both sugars concurrently without experiencing carbon catabolite repression. Production of 2,3-butanediol is demonstrated to characterize chemical production from the sugars in lignocellulosic hydrolysates. The engineered strain achieves a titer of 13.5 g L −1 of 2,3-butanediol over 12 days under shake-flask conditions. This study can be used as a foundation for industrial scale production of commodity chemicals from a combination of sunlight, CO 2 , and lignocellulosic sugars.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Production of renewable alcohols from maple wood using supercritical methanol hydrodeoxygenation in a semi-continuous flowthrough reactor

Biomass conversion to alcohols using supercritical methanol depolymerization and hydrodeoxygenation (SCM-DHO) with CuMgAl mixed metal oxide is a promising process for biofuel production. Here, we demonstrate how maple wood can be converted at high weight loadings and product concentrations in a batch and a semi-continuous reactor to a mixture of C 2 –C 10 linear and cyclic alcohols. Maple wood was solubilized semi-continuously in supercritical methanol and then converted to a mixture of C 2 –C 9 alcohols and aromatics over a packed bed of CuMgAlO x catalyst. Up to 95 wt% of maple wood can be solubilized in the methanol by using four temperature holds at 190, 230, 300, and 330 °C. Lignin was solubilized at 190 and 230 °C to a mixture of monomers, dimers, and trimers while hemicellulose and cellulose solubilized at 300 and 330 °C to a mixture of oligomeric sugars and liquefaction products. The hemicellulose, cellulose, and lignin were converted to C 2 –C 10 alcohol fuel precursors over a packed bed of CuMgAlO x catalyst with 70–80% carbon yield of the entire maple wood. The methanol reforming activity of the catalyst decreased by 25% over four beds of biomass, which corresponds to 5 turnovers for the catalyst, but was regenerable after calcination and reduction. In batch reactions, maple wood was converted at 10 wt% in methanol with 93% carbon yield to liquid products. The product concentration can be increased to 20 wt% by partially replacing the methanol with liquid products. The yield of alcohols in the semi-continuous reactor was approximately 30% lower than in batch reactions likely due to degradation of lignin and cellulose during solubilization. These results show that solubilization of whole biomass can be separated from catalytic conversion of the intermediates while still achieving a high yield of products. However, close contact of the catalyst and the biomass during solubilization is critical to achieve the highest yields and concentration of products.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2020

Climate change and the future productivity and distribution of crab in the Bering Sea

Crab populations in the eastern Bering Sea support some of the most valuable fisheries in the United States, but their future productivity and distribution are uncertain. Here, we explore observed changes in the productivity and distribution for snow crab, Tanner crab, and Bristol Bay red king crab. We link historical indices of environmental variation and predator biomass with observed time series of centroids of abundance and extent of crab stock distribution; we also fit stock–recruit curves including environmental indices for each stock. We then project these relationships under forcing from global climate models to forecast potential productivity and distribution scenarios. Our results suggest that the productivity of snow crab is negatively related to the Arctic Oscillation (AO) and positively related to ice cover; Tanner crab’s productivity and distribution are negatively associated with cod biomass and sea surface temperature. Aspects of red king crab distribution and productivity appear to be related to bottom temperature, ice cover, the AO, and/or cod biomass. Projecting these relationships forward with available forecasts suggests that Tanner crab may become more productive and shift further offshore, red king crab distribution may contract and move north, and productivity may decrease for snow crab as the population contracts northward.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2024

Economic and environmental bottlenecks in the industrial‐scale production of lipid‐derived biofuels from oleaginous yeasts: A review of the current trends and future prospects

Abstract Concerns about climate change and the reliance on fossil fuel reserves have motivated researchers to identify new renewable sources of energy. Biomass holds the potential to replace fossil‐derived products with biofuels and bio‐based chemicals. Plant‐derived lipids are promising sources of biofuels; however, the production of plant oil often leads to the release of massive amounts of carbon dioxide due to deforestation and land‐use change. The production of biofuels via plant oils (such as soybean) also competes with food production and in turn, impacts biodiversity. To mitigate these issues, the production of lipids from oleaginous yeasts could be an excellent alternative by incorporating these microbes into biorefineries utilizing agricultural or forest residues. Eventually, these microbial lipids could be potential sources for producing lipid‐derived biofuels. However, the current conventional methods for the production and recovery of lipids from oleaginous microbes suffer from economic and ecological challenges that affect its industrial‐scale expansion. This review highlights the major economic and environmental bottlenecks for the production of lipid‐derived biofuels from oleaginous yeasts. It also provides perspectives on the strategies that could be adapted on economic and ecological fronts to assist the expansion of the production of microbial lipid‐derived biofuels at an industrial scale.

Agriculture↗

DOE OSTI2024

Metabolic engineering of Caldicellulosiruptor bescii for 2,3-butanediol production from unpretreated lignocellulosic biomass and metabolic strategies for improving yields and titers

ABSTRACT The platform chemical 2,3-butanediol (2,3-BDO) is used to derive products, such as 1,3-butadiene and methyl ethyl ketone, for the chemical and fuel production industries. Efficient microbial 2,3-BDO production at industrial scales has not been achieved yet for various reasons, including product inhibition to host organisms, mixed stereospecificity in product formation, and dependence on expensive substrates (i.e., glucose). In this study, we explore engineering of a 2,3-BDO pathway inCaldicellulosiruptor bescii, an extremely thermophilic (optimal growth temperature = 78°C) and anaerobic bacterium that can break down crystalline cellulose and hemicellulose into fermentable C 5 and C 6 sugars. In addition, C. besciigrows on unpretreated plant biomass, such as switchgrass. Biosynthesis of 2,3-BDO involves three steps: two molecules of pyruvate are condensed into acetolactate; acetolactate is decarboxylated to acetoin, and finally, acetoin is reduced to 2,3-BDO.C. besciinatively produces acetoin; therefore, in order to complete the 2,3-BDO biosynthetic pathway,C. besciiwas engineered to produce a secondary alcohol dehydrogenase (sADH) to catalyze the final step. Two previously characterized, thermostable sADH enzymes with high affinity for acetoin, one from a bacterium and one from an archaeon, were tested independently. When either sADH was present inC. bescii,the recombinant strains were able to produce up to 2.5-mM 2,3-BDO from crystalline cellulose and xylan and 0.2-mM 2,3-BDO directly from unpretreated switchgrass. This serves as the basis for higher yields and productivities, and to this end, limiting factors and potential genetic targets for further optimization were assessed using the genome-scale metabolic model ofC. bescii. IMPORTANCE Lignocellulosic plant biomass as the substrate for microbial synthesis of 2,3-butanediol is one of the major keys toward cost-effective bio-based production of this chemical at an industrial scale. However, deconstruction of biomass to release the sugars for microbial growth currently requires expensive thermochemical and enzymatic pretreatments. In this study, the thermo-cellulolytic bacteriumCaldicellulosiruptor besciiwas successfully engineered to produce 2,3-butanediol from cellulose, xylan, and directly from unpretreated switchgrass. Genome-scale metabolic modeling ofC. besciiwas applied to adjust carbon and redox fluxes to maximize productivity of 2,3-butanediol, thereby revealing bottlenecks that require genetic modifications.

Biotechnology & Applied Microbiology↗

DOE OSTI2025

Alternate routes to acetate tolerance lead to varied isoprenol production from mixed carbon sources in Pseudomonas putida

ABSTRACT Lignocellulose is a renewable resource for the production of a diverse array of platform chemicals, including the biofuel isoprenol. Although this carbon stream provides a rich source of sugars, other organic compounds, such as acetate, can be used by microbial hosts. Here, we examined the growth and isoprenol production in a Pseudomonas putida strain pre-tolerized (“PT”) background where its native isoprenol catabolism pathway is deleted, using glucose and acetate as carbon sources. We found that PT displays impaired growth in minimal medium containing acetate and often fails to grow in glucose-acetate medium. Using a mutant recovery-based approach, we generated tolerized strains that overcame these limitations, achieving fast growth and isoprenol production in the mixed carbon feed. Changes in the glucose and acetate assimilation routes, including an upregulation in PP_0154 (SpcC, succinyl-CoA:acetate CoA-transferase) and differential expression of the gluconate assimilation pathways, were key for higher isoprenol titers in the tolerized strains, whereas a different set of mechanisms were likely enabling tolerance phenotypes in media containing acetate. Among these, a coproporphyrinogen-III oxidase (HemN) was upregulated across all tolerized strains and in one isolate required for acetate tolerance. Utilizing a defined glucose and acetate mixture ratio reflective of lignocellulosic feedstocks for isoprenol production in P. putida allowed us to obtain insights into the dynamics and challenges unique to dual carbon source utilization that are obscured when studied separately. Together, this enabled the development of a P. putida bioconversion chassis able to use a more complex carbon stream to produce isoprenol. IMPORTANCE Acetate is a relatively abundant component of many lignocellulosic carbon streams and has the potential to be used together with sugars, especially in microbes with versatile catabolism such as P. putida . However, the use of mixed carbon streams necessitates additional optimization. Furthermore, the use of P. putida for the production of the biofuel target, isoprenol, requires the use of engineered strains that have additional growth and production constraints when cultivated in acetate and glucose mixtures. In this study, we generate acetate-tolerant P. putida strains that overcome these challenges and examine their ability to produce isoprenol. We show that acetate tolerance and isoprenol production, although independent phenotypes, can both be optimized in a given P. putida strain. Using proteomics and whole genome sequencing, we examine the molecular basis of both phenotypes and show that tolerance to acetate can occur via alternate routes and result in different impacts on isoprenol production.

de Siqueira, Guilherme M. V. (ORCID:00000002364563↗

DOE OSTI2022

Establishing Butyribacterium methylotrophicum as a Platform Organism for the Production of Biocommodities from Liquid C 1 Metabolites

Using the Wood-Ljungdahl pathway, acetogens can nonphotosynthetically fix gaseous C 1 molecules, preventing them from entering the atmosphere. Many acetogens can also grow on liquid C 1 compounds such as formate and methanol, which avoid the storage and mass transfer issues associated with gaseous C 1 compounds. Substrate redox state also plays an important role in acetogen metabolism and can modulate products formed by these organisms. Butyribacterium methylotrophicum is an acetogen known for its ability to synthesize longer-chained molecules such as butyrate and butanol, which have significantly higher values than acetate or ethanol, from one-carbon (C 1 ) compounds. We explored B. methylotrophicum’s C 1 metabolism by varying substrates, substrate concentrations, and substrate feeding strategies to improve four-carbon product titers. Our results showed that formate utilization by B. methylotrophicum favored acetate production and methanol utilization favored butyrate production. Cofeeding of both substrates produced a high butyrate titer of 4 g/liter when methanol was supplied in excess to formate. Testing of formate feeding strategies, in the presence of methanol, led to further increases in the butyrate to acetate ratio. Mixotrophic growth of liquid and gaseous C 1 substrates expanded the B. methylotrophicum product profile, as ethanol, butanol, and lactate were produced under these conditions. We also showed that B. methylotrophicum is capable of producing caproate, a six-carbon product, presumably through chain elongation cycles of the reverse β-oxidation pathway. Furthermore, we demonstrated butanol production via heterologous gene expression. Finally, our results indicate that both selection of appropriate substrates and genetic engineering play important roles in determining titers of desired products.

09 BIOMASS FUELS↗

DOE OSTI2023

Measurements of Higgs boson production in the decay channel with a pair of τ leptons in proton–proton collisions at $\sqrt{s}=13$ TeV

Measurements of Higgs boson production, where the Higgs boson decays into a pair of τ leptons, are presented, using a sample of proton-proton collisions collected with the CMS experiment at a center-of-mass energy of , corresponding to an integrated luminosity of 138 fb –1 . Three analyses are presented. Two are targeting Higgs boson production via gluon fusion and vector boson fusion: a neural network based analysis and an analysis based on an event categorization optimized on the ratio of signal over background events. These are complemented by an analysis targeting vector boson associated Higgs boson production. Results are presented in the form of signal strengths relative to the standard model predictions and products of cross sections and branching fraction to τ leptons, in up to 16 different kinematic regions. For the simultaneous measurements of the neural network based analysis and the analysis targeting vector boson associated Higgs boson production signal strengths are found to be 0.82 ± 0.11 for inclusive Higgs boson production, 0.67 ± 0.19 (0.81 ± 0.17) for the production mainly via gluon fusion (vector boson fusion), and 1.79 ± 0.45 for vector boson associated Higgs boson production.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗

DOE OSTI2023

Projecting Future Energy Production from Operating Wind Farms in North America. Part III: Variability

Abstract Daily expected wind power production from operating wind farms across North America are used to evaluate capacity factors (CF) computed using simulation output from the Weather Research and Forecasting (WRF) Model and to condition statistical models linking atmospheric conditions to electricity production. In Parts I and II of this work, we focus on making projections of annual energy production and the occurrence of electrical production drought. Here, we extend evaluation of the CF projections for sites in the Northeast, Midwest, southern Great Plains (SGP), and southwest U.S. coast (SWC) using statewide wind-generated electricity supply to the grid. We then quantify changes in the time scales of CF variability and the seasonality. Currently, wind-generated electricity is lowest in summer in each region except SWC, which causes a substantial mismatch with electricity demand. While electricity of residential heating may shift demand, research presented here suggests that summertime CF are likely to decline, potentially exacerbating the offset between seasonal peak power production and current load. The reduction in summertime CF is manifest for all regions except the SGP and appears to be linked to a reduction in synoptic-scale variability. Using fulfillment of 50% and 90% of annual energy production to quantify interannual variability, it is shown that wind power production exhibits higher (earlier fulfillment) or lower (later fulfillment) production for periods of over 10–30 years as a result of the action of internal climate modes. Significance Statement Electrical power system reassessment and redesign may be needed to aid efficient increased use of variable renewables in the generation of electricity. Currently wind-generated electricity in many regions of North America exhibits a minimum in summertime and hence is not well synchronized with electricity demand, which tends to be maximized in summer. Future projections indicate evidence of reductions in wind power during summer that would amplify this offset. However, electrification of heating may lead to increased wintertime demand, which would lead to greater synchronization.

Coburn, Jacob↗

DOE OSTI2020

Metabolic engineering of Zymomonas mobilis for anaerobic isobutanol production

Biofuels and value-added biochemicals derived from renewable biomass via biochemical conversion have attracted considerable attention to meet global sustainable energy and environmental goals. Isobutanol is a four-carbon alcohol with many advantages that make it attractive as a fossil-fuel alternative. Zymomonas mobilis is a highly efficient, anaerobic, ethanologenic bacterium making it a promising industrial platform for use in a biorefinery. In this study, the effect of isobutanol on Z. mobilis was investigated, and various isobutanol-producing recombinant strains were constructed. The results showed that the Z. mobilis parental strain was able to grow in the presence of isobutanol below 12 g/L while concentrations greater than 16 g/L inhibited cell growth. Integration of the heterologous gene encoding 2-ketoisovalerate decarboxylase such as kdcA from Lactococcus lactis is required for isobutanol production in Z. mobilis. Moreover, isobutanol production increased from nearly zero to 100-150 mg/L in recombinant strains containing the kdcA gene driven by the tetracycline-inducible promoter Ptet. In addition, we determined that overexpression of a heterologous als gene and two native genes (ilvC and ilvD) involved in valine metabolism in a recombinant Z. mobilis strain expressing kdcA can divert pyruvate from ethanol production to isobutanol biosynthesis. This engineering improved isobutanol production to above 1 g/L. Finally, recombinant strains containing both a synthetic operon, als-ilvC-ilvD, driven by Ptet and the kdcA gene driven by the constitutive strong promoter, Pgap, were determined to greatly enhance isobutanol production with a maximum titer about 4.0 g/L. Finally, isobutanol production was negatively affected by aeration with more isobutanol being produced in more poorly aerated flasks. This study demonstrated that overexpression of kdcA in combination with a synthetic heterologous operon, als-ilvC-ilvD, is crucial for diverting pyruvate from ethanol production for enhanced isobutanol biosynthesis. Moreover, this study also provides a strategy for harnessing the valine metabolic pathway for future production of other pyruvate-derived biochemicals in Z. mobilis.

09 BIOMASS FUELS↗

DOE OSTI2025

Optimal production of Phanerochaete chrysosporium manganese peroxidases and Trametes sp. C30 laccase hybrid Lac131 in Aspergillus niger for lignin bioconversion

Background Incorporating the production of related ligninolytic enzymes into industrial filamentous fungus Aspergillus niger will enhance the bioconversion of lignocelluloses to various chemical products. Results In this study, transgenic expression of Phanerochaete chrysosporium manganese peroxidases (mnps) and Trametes sp. C30 laccase hybrid Lac131 (lac131) were examined and optimized in A. niger 11414 prtT∆ strain. Five mnps (mnp1, mnp2, mnp3, mnp4, and mnp5) and lac131 genes were expressed separately or in combination. The transgenic strain containing the entire mnp2 genomic coding sequence (gmnp2) exhibited the highest mnP activity among the five mnp over-expression strains in the modified minimal medium (mMM) with addition of 5 g/L bovine hemoglobin (bHg). We examined the effects of hemin and bHg on mnP production in the gmnp2 strain cultures and found that at least 1 g/L bHg was required, while hemin was not. Culture conditions for mnP production were further optimized for the gmnp2 strain and the highest mnP activities were detected in the cultures grown at 25 °C and 200 rpm with an initial pH of 4.5. Effects of soy protein, skim milk, and bovine serum albumin on mnP production were investigated; 5 g/L of soy proteins or skim milk had comparable effects to 2.5 g/L bHg, while cultures with bovine serum albumin had diminished mnP activity. Disruption of both prtT and vsm1 substantially augmented the mnP production and its activity reached 575 U/L. Trametes sp. C30 laccase hybrid lac131 was strongly expressed in either A. niger gmnp2 (1975 U/L) or 11414prtT∆ (3895 U/L) strain. Both mnP and laccase in the culture supernatants effectively decolorized selected phenolic compounds (dyes) and cleaved tagged model lignin dimers. Conclusion The mnP was successfully produced in A. niger by optimizing the culture conditions and host strain. Co-expression of all four mnp genes in the same expression host by multiplex CRISPR will lead to the mnP production reaching levels comparable to P. chrysosporium, while only requiring 36 h at 25 °C. The Lac131 activity in transgenic A. niger strain is 4- to 7-times higher than that in previous studies. Co-production of mnP and laccase in A. niger will enhance the lignin bioconversion efficiency.

Aspergillus niger↗