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All sourcesNASA NTRSDOE OSTI
DOE OSTI2021

Enhanced medium chain length-polyhydroxyalkanoate production by co-fermentation of lignin and holocellulose hydrolysates

Biological lignin conversion to medium chain length-polyhydroxyalkanoates (mcl-PHA) has recently emerged as an attractive alternative to petroleum-based plastics due to the renewable nature of lignin and the value-added applications of mcl-PHA. Previous reports suggested that addition of limited glucose can improve mcl-PHA accumulation in Pseudomonas putida KT2440 grown in lignin substrates. Herein, we propose a biorefinery process to systematically release lignin and sugars from lignocellulosic biomass and evaluate the potential of co-utilization of all biomass components for conversion to mcl-PHA. Our results indicate that a sequential treatment composed of acid pretreatment, enzymatic hydrolysis and alkaline treatment produces a suitable lignin stream for mcl-PHA production in engineered P. putida. In addition, the PHA titer could be increased by 71% when the lignin stream (AH) was combined with the enzymatic hydrolysate (EH) at a ratio of 75 : 25. Higher ratios of EH : AH negatively affect mcl-PHA accumulation as well as mixtures with sugars from the acid hydrolysate. The optimization of the fermentation conditions, including the inoculum (OD 600 ) and substrate (soluble solid content (SSC)), was carried out by a central composite design. The optimal conditions were achieved at OD 600 = 2.17 and SSC = 68.28 g L –1 . Under these conditions, the titer increased 4.3 fold, achieving a mcl-PHA production of 1.38 g L –1 . Lignin characterization before and after fermentation by nuclear magnetic resonance and gas chromatography showed that p-coumarates were mainly consumed during the fermentation for mcl-PHA production. Overall, this biorefinery strategy allowed us to increase mcl-PHA production by utilizing the different lignocellulosic fractions. Unlike previous studies, no model substrates were utilized at any stage of the fermentation process, representing a step forward towards process feasibility.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2022

Plasma technology for lignocellulosic biomass conversion toward an electrified biorefinery

Lignocellulosic biomass conversion to renewable, carbon-neutral materials, fuels, and chemicals is the cornerstone of the transition to a sustainable future bioeconomy. Green energy in the form of electricity needs to be coupled with or substitute conventional thermally driven processes to realize small-scale, economically viable and environmentally friendly biorefineries. Gas discharge plasmas enable the conversion of renewable electric energy, supplied in the form of an electric field, to chemical energy through the formation of a highly reactive environment that can induce several transformations related to agricultural waste valorization processes. Herein, we review the application of plasma technology to lignocellulosic biomass upgrade, aiming to provide the scientific background and technical challenges in this rapidly emerging research field. To bridge the gap between plasma science and biomass valorization technologies, we initially present the technical aspects of plasma reactors related to biomass processing and further discuss the advances in plasma processing for each biomass conversion technology, providing insights into the related plasma chemistry and interaction mechanisms. Here, we first focus on the low and medium-temperature biomass conversion processes, including biomass pretreatment and delignification to promote enzyme or acid-catalyzed hydrolysis to sugars and biomass liquefaction using plasma electrolysis. Then we discuss the high and very high-temperature conversion processes, such as plasma-assisted pyrolysis and gasification to syngas and plasma application to tar removal, combustion, and vitrification. Overall, this review provides knowledge at the interface of plasma science and biomass conversion technology to promote the interaction between the individual communities, which is crucial for the further advancement of the field.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Prediction of solubility parameters of lignin and ionic liquids using multi-resolution simulation approaches

The solubility parameter (SP) of a molecular species is a vital feature that indicates polarity and quantifies the ‘like-seeks-like’ principle, which is used in chemistry to screen solvents for dissolution. Recent studies have demonstrated that ionic liquids (ILs) and deep eutectic solvents (DESs) efficiently solubilize lignocellulosic biomass and promote enzymatic saccharification into sugars used for the production of biofuels and value-added chemicals. Understanding the solubility of plant biopolymers, particularly lignin, in ILs and DESs is critical for selecting candidate ILs and DESs for biomass pretreatment; however, experimentally measuring SPs is challenging. Thus, the present study investigates lignin dissolution mechanisms in IL/DES and prediction of the solubility parameters (Hildebrand and Hansen) of lignin, ILs, and DESs using multi-resolution simulation approaches. Here, solubility parameters of the studied compounds were predicted using molecular dynamics (MD) simulations, and the SP of lignin was determined to be 23–27 MPa 1/2 , which was close to the polymeric lignin solubility parameter (24.3–25.5 MPa 1/2 ). The SPs of ILs namely [Ch][Lys], [Ch][Oct], and [Emim][Lys] were predicted to be ~26 MPa 1/2 , which is close to lignin's SPs and resulted in increased biomass delignification. The MD simulated SPs were validated by both the COSMO-RS model and experimental investigations, with the results showing a close agreement between the predicted and experimentally obtained SPs. In addition, the enthalpy of vaporization (ΔH vap ) of ILs/DESs was predicted based on the potential energy of the system, and the ΔH vap of ILs/DESs was around 40–65 kcal mol –1 , which is 5–8 times higher than that of traditional organic solvents.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2023

Enhanced microbial production of protocatechuate from engineered sorghum using an integrated feedstock-to-product conversion technology

Building a stronger bioeconomy requires production capabilities that are largely generated through microbial genetic engineering. Plant feedstocks can additionally be genetically engineered to generate desirable feedstock traits and provide precursors for direct microbial conversion into desired products. The oleaginous yeast Rhodosporidium toruloides is a promising organism for this type of conversion as it can grow on a wide range of deconstructed biomass and consume a variety of carbon sources. Here, we leveraged R. toruloides native p-coumaric acid consumption pathway to accumulate protocatechuate (PCA) from 4-hydroxybenzoate (4HBA) released from a sorghum feedstock line genetically engineered to overproduce 4HBA. We did so by generating and evaluating an R. toruloides strain that accumulates PCA, RSΔ12623. We then show that at two scales a cholinium lysinate pretreatment with enzymatic saccharification successfully extracts 95% of the 4HBA from the engineered sorghum biomass while producing deconstructed lignin that can be more efficiently depolymerized in a subsequent thermochemical reaction. We also demonstrate that strain RSΔ12623 can convert more than 95% of 4HBA to PCA while consuming >95% of the glucose and >80% of the xylose present in sorghum hydrolysates. Finally, to evaluate the scalability of such fermentations, we conducted the conversion of 4HBA to PCA in a 2 L bioreactor under controlled conditions. Importantly, this work demonstrates the potential of purposefully producing aromatic precursors in planta that can be liberated during biomass deconstruction for direct microbial conversion to desirable bioproducts.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2024

Solid–liquid separation of lignocellulosic sugars from biomass by rotating ceramic disc filtration

In many biomass conversion processes, the separation of cellulosic sugars from residual, lignin-rich solids is a critical step, and achieving high recovery yields of sugars by conventional tangential crossflow and vacuum filtration is challenged by the presence of biomass solids, which rapidly foul filters, resulting in decreased throughput. Considering the performance limitations of traditional filtration methods, dynamic filtration, which generates high shear at the membrane surface to decrease fouling, is emerging as a viable alternative for demanding solid–liquid separations. For high solids separations, there is little available information regarding the performance, limitations, and energy consumption of dynamic filtration. To that end, here we characterized the performance of a dynamic filtration module, specifically a rotating ceramic disc (RCD) filter, for the aseptic recovery of cellulosic sugars from biomass solids following pretreatment and enzymatic hydrolysis. We show how RCD rotational velocity and percent biomass solids impact the filter throughput. Additionally, we used computational fluid dynamics (CFD) simulations to estimate the shear rate at the membrane surface and to visualize hydrodynamic profiles within the module. With the combined CFD simulations and experimental results, we estimated the energy demand and operating expenses for a viable dynamic filtration system operating with a lignocellulosic feed slurry. Our results indicate that an RCD filter can achieve ≥95% recovery of sugars and produce a retentate slurry containing 12 wt% insoluble solids with low energy consumption (a 2.2-fold improvement over cross-flow filtration) and low operating costs ($\$$0.06 per kg sugars). These results show a viable path towards operationally reliable, energy efficient, and cost-effective separations of sterilized cellulosic sugars from biomass solids and highlight the potential of dynamic filtration systems for challenging solid–liquid separations.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Maximizing long-term biohydrogen production with Clostridium thermocellum for high solids conversion of lignocellulosic biomass

Biological hydrogen production from lignocellulosic biomass sustainably couples organic waste reduction with renewable energy generation. Efficient conversion is challenged by the structural complexity of lignocellulose and resulting recalcitrance to enzymatic degradation. Clostridium thermocellum natively breaks down biomass with highly effective hemi-/cellulases systems (i.e., cellulosomes) and generates hydrogen in anaerobic cultivation, creating a compelling platform for lignocellulosic biohydrogen production. Achieving commercially viable production rates requires balancing high biomass loading and throughput against uniform mixing conditions required for enzyme dispersion, pH and temperature control, and efficient hydrogen and metabolite removal in continuous operation. To address these barriers to process intensification, we implemented novel reactor and process designs for high-solids lignocellulosic biomass fermentations using the C. thermocellum KJC19-9 strain, genetically engineered for co-utilization of cellulose and hemicellulose sugars (i.e., xylose). Via computational fluid dynamics (CFD) modeling and experimental validation, we achieved a >50% improvement in biohydrogen production with an improved anchor-type impeller morphology, coupled to a threefold reduction in agitation rate. To further reduce rheological constraints and accumulation of toxic metabolites, we then transitioned the process to sequencing fed-batch operation. The resulting process generated 24.87 L H 2 L −1 from 160 g L −1 of deacetylated and mechanically refined (DMR)-pretreated corn stover biomass over 16 days while solubilizing >95% of influent cellulose and hemicellulose, setting a new performance benchmark for continuous production of biohydrogen from lignocellulose.

08 HYDROGEN↗
DOE OSTI2026

Energy utilization strategies in lignocellulosic biorefineries

Lignocellulosic biorefineries can generate various energy-rich side streams, including biogas from anaerobic digestion, a lignin stream from biomass pretreatment, and conversion residues. This study evaluates different chemical energy utilization strategies in lignocellulosic biorefineries based on energy and carbon efficiencies, cost, and greenhouse gas mitigation potential. Specifically, using modeling and optimization, we examine strategies that include electricity generation, biogas upgrade to biomethane, lignin valorization, and carbon capture and storage. We find that upgrading biogas to biomethane demonstrates the highest energy efficiency and leads to the lowest minimum fuel selling price for the main product, ethanol. The biorefinery with carbon capture and storage achieves the lowest net carbon footprint. Lignin valorization has the highest carbon footprint due to the additional materials required for lignin depolymerization, despite its potential to produce high value bioproducts.

Aboagye, Emmanuel A. [Princeton Univ., NJ (United ↗
DOE OSTI2026

Selective designer hydrolysates from Miscanthus bioenergy crop for sustainable utilization

Designer enzymatic hydrolysates deliver targeted, optimized fermentable sugars while reducing the formation of inhibitory by-products. Herein, five distinct enzymatic hydrolysates (EH) were generated from Miscanthus biomass using alkaline-ethanol fractionation and hydrothermal-mechanical refining (HMR) pretreatment approaches. Noticeably, alkali-ethanol fractionated hemicellulose A (HMA) and hemicellulose B (HMB) hydrolysis resulted in xylose-rich enzymatic hydrolysates with 104.8 ± 5.6 g L −1 and 173.4 ± 4.6 g L −1 xylose, respectively. All five hydrolysates encompassing varied carbon to nitrogen ratios were evaluated for lipid production using an engineered oleaginous yeast, R. toruloides 880-ADS. Microbial fermentation of xylose-rich hydrolysates, viz. HMR-EH2, HMA-EH, and HMB-EH, resulted in lipid yields ranging from 0.05 to 0.12 g g −1 sugar. Compositional and 2D NMR analyses revealed that HMA-EH residues are highly lignin-enriched while preserving the non-condensed structure conducive to valorization. Overall, the findings highlighted a novel, selective trade-off approach for maximizing lipid titers from designer hydrolysates, corroborating complete resource recovery for establishing integrated biorefineries.

Singh, Shuchi [University of Illinois at Urbana-Ch↗
DOE OSTI2023

Thermodynamic Properties of Aggregate Coated by Different Types of Waste Plastic: Adhesion and Moisture Resistance of Asphalt-Aggregate Systems

The utilization of acidic aggregates in hot-mix asphalt (HMA) is often inevitable due to the limited supply of locally available aggregates, posing a potential threat of moisture damage to asphalt pavements. In this regard, the use of urban waste plastic to coat the low-quality aggregates is a promising way to improve the moisture resistance. This study fundamentally evaluated the compatibility between asphalt and plastic-coated aggregate by measuring surface energy, and for the first time compared the effects of different types of waste plastic including high-density polyethylene (HDPE), low-density polyethylene (LDPE), polyethylene terephthalate (PET), polyvinyl chloride (PVC), and polypropylene (PP). The plastic-coating significantly altered the chemistry of aggregate surface by increasing the nonpolar components and reducing the polar components. Such changes remarkably increased the dry adhesion energy between asphalt and aggregate and enhanced the wettability of aggregate. For the acidic aggregates, the polymeric treatment could decrease the value of wet adhesive, which indicated the debonding tendency in the presence of water was reduced. Finally, the energy ratios of the asphalt-aggregate combinations significantly increased by the surface pretreatment, which proved the effect of waste plastic coating on improving the moisture damage resistance of asphalt mixtures. By comparison, the HDPE coating was the most effective way to enhance the compatibility with asphalt, followed by PP and LDPE coatings. The PVC and PET coatings were investigated as the least effective way. In conclusion, this study cannot only contribute to the potential large-scale utilization of waste plastic but also remove the obstacles of using large quantities of acidic natural aggregate in asphalt pavements.

36 MATERIALS SCIENCE↗
DOE OSTI2021

On the characteristics of N-polar GaN Schottky barrier contacts with LPCVD SiN interlayers

In this work, we study the behavior of N-polar GaN Schottky diodes with low-pressure chemical vapor deposited (LPCVD) SiN interlayers and unveil the important role of an amphoteric miniband formed in this interlayer due to a previously identified and dominating Si dangling bond defect. Through analysis of temperature-dependent current–voltage (I–V–T), capacitance–voltage (C–V), and x-ray photoelectron spectroscopy measurements, we observe that when nickel is deposited on LPCVD SiN pretreated with hydrofluoric acid, the SiN/GaN interface is responsible for determining the overall system's barrier height. By contrast, contact formation on oxidized LPCVD SiN leads to a metal/SiN-dominant barrier. We, consequently, propose band diagrams that account for an amphoteric miniband in LPCVD SiN, leading to a new understanding of LPCVD SiN as a lossy dielectric with surface barrier-dependent behavior.

42 ENGINEERING↗
DOE OSTI2021

A multi-omics approach to lignocellulolytic enzyme discovery reveals a new ligninase activity from Parascedosporium putredinis NO1

Lignocellulose, the structural component of plant cells, is a major agricultural byproduct and the most abundant terrestrial source of biopolymers on Earth. The complex and insoluble nature of lignocellulose limits its conversion into value-added commodities, and currently, efficient transformation requires expensive pretreatments and high loadings of enzymes. In this work, we report on a fungus from the Parascedosporium genus, isolated from a wheat-straw composting community, that secretes a large and diverse array of carbohydrate-active enzymes (CAZymes) when grown on lignocellulosic substrates. We describe an oxidase activity that cleaves the major β-ether units in lignin, thereby releasing the flavonoid tricin from monocot lignin and enhancing the digestion of lignocellulose by polysaccharidase mixtures. We show that the enzyme, which holds potential for the biorefining industry, is widely distributed among lignocellulose-degrading fungi from the Sordariomycetes phylum.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Ion Exchange of Selected Group II Metals and Lead by Crystalline Silicotitanate and Competition for Cs Exchange Sites

A series of batch contact tests were conducted to evaluate the exchange behavior of Ba, Ca, Pb, and Sr onto crystalline silicotitanate (CST) in support of an expedited Cs removal and pretreatment system at the Hanford site. Binary Na/M 2+ and ternary Na/Cs/M 2+ isotherms were generated to understand selectivity, capacity, and competitive impact of each analyte on Cs uptake from a simple 1 M NaOH/4.6 M NaNO 3 simulant. Analyte loading from a 0.1 M NaOH/5.5 M NaNO 3 simulant was assessed to determine the effect of hydroxide concentration on binary Na/M 2+ isotherms. Finally, results from binary and ternary isotherms indicated that group II metals, and Pb do not impact CST performance toward CST at concentrations expected in Hanford tank waste supernate.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Small to Full-Height Scale Comparisons of Cesium Ion Exchange Performance with Crystalline Silicotitanate

Crystalline silicotitanate (CST) ion exchanger is planned to be used to remove cesium (137Cs) from the aqueous phases of Hanford tank wastes in preparation for vitrification at the Waste Treatment and Immobilization Plant (WTP) Pretreatment Facility. Column scale up testing was conducted to evaluate performance of Cs exchange onto the CST. Batch contact testing was conducted to assess exchange kinetics of Cs exchange at four different CST PSDs. Column testing was conducted at three column sizes, small (2.5% full height), medium (12% full height), and full height, to assess Cs load performance behavior. Testing at the small scale was compared to actual Hanford tank waste testing to verify the validity of simulant tests to accurately represent full height column performance. A change in CST particle size was essential in scaling the small column dynamics up to full scale. Based on these results, a determination of intraparticle and film diffusion impacts on overall mass transfer coefficients will allow future modeling of the breakthrough performance at a range of process conditions.

Westesen, Amy M.↗
DOE OSTI2021

Mg x Zn 1−x O contact to CuGa 3 Se 5 absorber for photovoltaic and photoelectrochemical devices

Abstract CuGa 3 Se 5 is a promising candidate material with wide band gap for top cells in tandem photovoltaic and photoelectrochemical (PEC) devices. However, traditional CdS contact layers used with other chalcopyrite absorbers are not suitable for CuGa 3 Se 5 due to the higher position of its conduction band (CB) minimum. Mg x Zn 1− x O (MZO) is a transparent oxide with adjustable band gap and CB position as a function of magnesium composition, but its direct application is hindered by CuGa 3 Se 5 surface oxidation. Here, MZO is investigated as a contact (n-type ‘buffer’ or ‘window’) material to CuGa 3 Se 5 absorbers pretreated in Cd 2+ solution, and an onset potential close to 1 V vs reversible hydrogen electrode in 10 mM hexaammineruthenium (III) chloride electrolyte is demonstrated. The Cd 2+ surface treatment changes the chemical composition and electronic structure of the CuGa 3 Se 5 surface, as demonstrated by photoelectron spectroscopy measurements. The performance of CuGa 3 Se 5 absorber with Cd 2+ treated surface in the solid-state test structure depends on the Zn/Mg ratio in the MZO layer. The measured open circuit voltage of 925 mV is promising for tandem PEC water splitting with CuGa 3 Se 5 /MZO top cells.

36 MATERIALS SCIENCE↗
DOE OSTI2021

Prolonging photoperiod promotes testosterone synthesis of Leydig cells by directly targeting local melatonin system in rooster testes

Abstract The study investigated the effects of prolonging photoperiod on the synthesis of testosterone and melatonin in roosters, and the effect of melatonin on testosterone synthesis in rooster Leydig cells as well as its molecular mechanisms. We randomly divided one hundred and twenty 20-week-old roosters into three groups and provided 6, 12.5 and 16 h light, respectively. The results showed that prolonging photoperiod promoted testosterone synthesis, decreased melatonin production, and inhibited the expression of melatonin membrane receptors MEL1A, MEL1B, MEL1C, and aralkylamine n-acetyltransferase (AANAT) in rooster testes. Subsequently, rooster Leydig cells were isolated and treated with 0, 0.1, 1, 10, and 100 ng/mL melatonin for 36 h. The results suggested that melatonin inhibited testosterone synthesis in rooster Leydig cells, and silencing MEL1A and MEL1B relieved the inhibition of melatonin on testosterone synthesis. Additionally, melatonin reduced the intracellular cyclic adenosine monophosphate (cAMP) level and the phosphorylation level of cAMP-response element binding protein (CREB), and CREB overexpression alleviated the inhibition of melatonin on testosterone synthesis. Furthermore, pretreatment with cAMP activator forskolin or protein kinase A (PKA) activator 8-bromo-cAMP blocked the inhibition of melatonin on CREB phosphorylation and testosterone synthesis. These results indicated that prolonging photoperiod promoted testosterone synthesis associated with the decrease in melatonin production and membrane receptors and biosynthetic enzyme of melatonin in rooster testes, and melatonin inhibited testosterone synthesis of rooster Leydig cells by inhibiting the cAMP/PKA/CREB pathway via MEL1A and MEL1B. This may be evidence that prolonging photoperiod could promote testosterone synthesis through the inhibition of the local melatonin pathway in rooster testes.

Xu, Gaoqing↗
DOE OSTI2021

Immunological Correlates of the HIV-1 Replication-Competent Reservoir Size

Understanding what shapes the latent human immunodeficiency virus type 1 (HIV-1) reservoir is critical for developing strategies for cure. Here, we measured frequency of persistent HIV-1 infection after 5 years of suppressive antiretroviral therapy initiated during chronic infection. Pretreatment CD8 + T-cell activation, nadir CD4 count, and CD4:CD8 ratio predicted reservoir size.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2025

Artemether-Lumefantrine Treatment Selects Plasmodium falciparum Multidrug Resistance 1 ( pfmdr1 ) Increased Copy Number Among African Malaria Infections

Abstract Background Decreased efficacy of artemether-lumefantrine, the globally most used antimalarial, has recently emerged in Africa. Methods An efficacy trial was carried out based on directly observed artemether-lumefantrine therapy at Bengo, Northern Angola. One-hundred Plasmodium falciparum uncomplicated malaria patients (2–10 years old) were enrolled, hospitalized for the treatment period, and followed up for 42 days. Polymerase chain reaction (PCR) correction was performed with pfmsp1/2 plus glurp, with analysis considering 2 or 3 coincident markers. Infections were tested by quantitative PCR (qPCR) for pfmdr1 copy number (pfmdr1×N), a potential P. falciparum marker of lumefantrine resistance previously identified in the region. In vitro clone mixtures were built and used to determine the relation between qPCR copy number scores and actual intrainfection quantitative fractions of pfmdr1×N. Results We observed a significant posttreatment selection of gene amplification, suggesting a role in the parasite in vivo response to this drug. pfmdr1×2 qPCR scores of 1.3, 1.4, and 1.5 were determined to correspond to 15%, 25%, and 35% intrainfection rates. Patients carrying infections with a score ≥1.4 at baseline were linked to decreased artemether-lumefantrine day 42 efficacy (79% vs 97% single-copy pfmdr1). All infections were pfmdr1 N86 carriers and no pfk13 mutations were found. Conclusions Our study suggests pfmdr1×N as a marker of P. falciparum in vivo response to lumefantrine in Africa, while indicating patients carrying infections with a pretreatment pfmdr1×N score ≥1.4 before treatment are a group experiencing decreased artemether-lumefantrine performance.

Fançony, Claudia (ORCID:0000000344211769)↗