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DOE OSTI2021

Editorial: Bioconversion and Biorefinery of C1 Compounds

The past decade has seen significant progress in the field of metabolic engineering and synthetic biology. The exponentially growing multi-omics data and technological advances in the development of efficient genetic manipulation tools and techniques have allowed scientists to explore and expand their understanding of microbial metabolisms and further develop sophisticated engineering strategies to realize the use of industrial "workhorses" and non-conventional microorganisms for sustainable bioconversion and biorefinery. There is of great interest for the research community in using C1 compounds (i.e., CO 2 , CO/syngas, methane, methanol) as the next generation feedstocks for microbial cell factories and biocatalysts to promote the sustainable development of a green economy (Figure 1). Considering lowering input costs is also a main consideration for successful business ventures, the use of inexpensive, abundant, and widely accessible C1 compounds is envisioned as a promising route for the sustainable production of fine chemicals, fuels, and other high-value products. Many C1 compounds are waste gases from industrial activities and may have detrimental effects on climate change upon emission into the atmosphere. Therefore, promoting the use of C1 compounds as renewable carbon feedstocks can greatly contribute to the reduction of anthropogenic emission of air pollutants.

09 BIOMASS FUELS↗

DOE OSTI2022

Designing Harvesting and Hauling Cost Models for Energy Cane Production for Biorefineries

The harvesting and hauling operations of bioenergy feedstock is an important area in biofuel production. Production costs can be minimized by maintaining optimal machinery units for these operations. The objective of this study is to design an optimal harvesting unit for bioenergy refinery and estimate harvesting and hauling costs of energy cane. A biorefinery with the annual capacity of processing twenty-five million imp. gallons of ethanol were considered. Given the efficiency of harvesting, a two-row soldier system was considered. Considering the year-round supply of energy cane to the refinery, the optimal machinery unit was designed, and the combined operation costs were derived. The average estimated ownership, repair, labor and fuel and lubricant costs of biomass harvest unit were calculated to be $\$$0.50, $\$$0.54, $\$$1.78 and $\$$1.51/mt, respectively. The costs distribution generated showed harvesting and hauling costs could range between $\$$5.47–$\$$9.23/mt of energy cane. The methodology and the research output will provide guidelines for investors in designing harvesting and hauling units and estimating costs for different scales of operation.

09 BIOMASS FUELS↗

DOE OSTI2021

VFA Biorefinery Design for Informed Production of Ground and Aviation Fuels

The rising demand for low-carbon intensity fuel options requires evaluation of a system which can support their production economically and sustainably. Volatile fatty acids (VFAs) ranging from C2 to C8 can be derived in high yield from arrested anaerobic digestion of biomass. This talk provides an overview of our work in which we upgrade wet waste-derived VFAs using ketonization to elongate their carbon backbone to reach a range relevant to jet or diesel fuels. Kinetic models were used to predict ketone profiles from varying VFA profiles, thereby informing potential carbon flow to either (a) mixed paraffins for use in diesel or aviation fuel, or (b) ethers for use in diesel fuel. To do this, the anticipated products were screened for critical fuel characteristics using predictive tools. The criteria for neat and blended bioblendstocks were chosen based on conventional petrofuel requirements, and they were applied to inform fuel targets, identify limiting characteristics, and guide conversion development. While paraffins can serve as either diesel or aviation fuels, the latter has stringent criteria which include a precise distillation curve range tied to carbon distribution. Fuels which fall outside this range typically fail to meet other property metrics, and as such flashpoint and viscosity were identified as potentially limiting properties of this VFA aviation fuel. However, paraffin fractions which fall outside aviation criteria may meet diesel fuel requirements, which are looser except for the flashpoint minimum. Flashpoint was also a concern for smaller VFA diesel ethers, which posed an additional oxygenate risk of high water solubility. This fuel-informed conversion design process was demonstrated through production of paraffins with reduced sooting as compared to petrodiesel (~34%), and increased renewable aviation fuel blend levels (>70%). VFA-derived ethers improved petrodiesel by increasing fuel autoignition quality by 56% and reducing sooting by 86%. These VFA aviation and diesel fuels could enable greenhouse gas (GHG) reductions of over 165% and 50%, respectively, relative to their purely fossil-derived counterparts. Collectively, this work provides (i) an overview of how we leveraged the flexibility of VFAs for fuel production; and (ii) insight into the potential of a VFA biorefinery to accommodate varied feed and fuel applications, while also considering the merit of tailored fuel production pathways and GHG reduction potential.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

DOE OSTI2022

Lignin-First Biorefinery Development

Cost-effective biomass fractionation is an enabling, grand challenge for biorefining, especially when both carbohydrates and lignin are targeted for valorization. The advent of Reductive Catalytic Fractionation (RCF) - an active stabilization approach that solubilizes lignin from biomass and catalytically depolymerizes it into a narrow slate of monomers and oligomers - represents a potential step forward for this important goal. In the Lignin-First Biorefinery Development project, we are employing techno-economic analysis (TEA) and life cycle assessment (LCA) to guide bench-scale R&D efforts towards cost-effective RCF-based biorefining. This project is critical to lignin valorization efforts and could enable the use of woody feedstocks in a traditional biochemical conversion context. To date, we have conducted the first rigorous TEA and LCA study of the RCF process, developed a flow-through system to separate biomass and the chemical catalyst in RCF chemistry, developed models for solvolysis chemistry and transport phenomena for poplar, and co-led an international, authoritative perspective on guidelines for the research community on how to best practice lignin-first biorefining. The primary challenges for the lignin-first RCF process going forward are catalyst stability, the need to utilize, recover, and recycle high boiling point solvents to lower RCF reactor pressure, and the challenge of operating RCF continuously - all of which are being directly tackled by this project.

biomass↗

DOE OSTI2025

Integrated Green Biorefinery for the Production of Anthocyanins, Fermentable Sugars, and High Pure Lignin from Miscanthus × giganteus

Miscanthus x giganteus (Mxg) is a promising perennial crop for producing natural colorants, renewable fuels, and bioproducts. However, natural recalcitrance and high pretreatment cost are major barriers to their complete conversion. In this study, a green processing method has been investigated for efficient recovery of natural pigments (anthocyanins), fermentable sugars, and pure lignin from Mxg genotypes using choline chloride-based natural deep eutectic solvents (NADES) systems. Interestingly, choline chloride: lactic acid (ChCl: LA) NADES-processed biomass resulted in 67.8 ± 2.1 μg g –1 of anthocyanins from dry biomass. A maximum of 87.4%–94.1% glucose yield was achieved after enzymatic saccharification. The effective extraction of lignin with high purity with higher β-aryl ether (β—O—4) bonds from advanced crops is crucial for lignin valorization. Notably, highly pure lignin (≈93.4% ± 1.4%) is achieved after low-temperature NADES pretreatment while retaining lignin's native structure. 31 P nuclear magnetic resonance demonstrated that total phenolics for ChCl: LA-lignin resulted in 1.20 mmol g –1 hydroxyls. The relative monolignol composition of syringyl (S), guaiacyl (G), and p-hydroxyphenyl (H) is 19.0, 65.7, and 14.3%, respectively, as evidenced by heteronuclear single quantum coherence analysis. This study provides a novel approach for obtaining high-purity lignin for catalytic depolymerization for oligomers and bifunctional monoaromatics production and leverages current cellulosic biorefinery technologies.

Raj, Tirath [University of Illinois at Urbana-Cham↗

DOE OSTI2023

Muconic acid production from algae hydrolysate as a high-value co-product of an algae biorefinery

Muconic acid is an attractive bio-derived product because it can be easily converted to adipic acid, a high-value and high-volume monomer used in the production of nylon and other valuable consumer plastics. As such, production of muconic acid from algae hydrolysate was explored to expand the suite of products available from algal biomass. Here we have established initial performance parameters and have shown that the range of substrates present in algae hydrolysate that are consumed by Pseudomonas putida includes at least glucose, mannose, glycerol, acetic acid, and lactic acid. We achieved complete utilization of these major carbon sources found in Scenedesmus acutus hydrolysate with a maximum muconic acid productivity of 0.25 g/L h. Final titer was 13.0 g/L (27% molar process yield) at approximately 50 h. In addition, algae hydrolysate does not require any additional nutrients to achieve these performance metrics. Techno-economic analysis showed the potential to support up to 150 algal biorefineries at this yield with significant greenhouse gas reductions.

09 BIOMASS FUELS↗

DOE OSTI2025

Enhancing the flowability of woody biomass slurries in wet biorefineries

Feeding wet lignocellulosic biomass (e.g., softwood and hardwood) slurries into high-pressure, high-temperature reactors at an industrially relevant scale presents significant challenges, such as process equipment plugging. Here, in this study, we investigate the possibility of improving the flowability of biomass slurries in an industrial scale wet biorefinery by tuning the physical and chemical characteristics of the biomass particles. To understand the effects of the chemical characteristics of woody biomass on flowability, cellulose pulp particles are produced from pulp sheets via knife milling, pelletizing, and crumbling. These cellulose pulp particles are processed for acid hydrolysis dehydration (AHDH) at one –tonne-per day pilot scale. The levulinic acid yield (a product of cellulose AHDH) and the flowability of the biomass particles are compared using market pulp, 1 mm crumbled pine softwood, a blend of 2 mm hammer-milled pine softwood with 10 wt.% bark, and 2 mm hammer-milled hardwood material with 10 wt.% bark (to represent forest residues). The results indicate that the presence of hemicellulose and lignin influences the flowability of lignocellulosic feedstocks. Crumbled wood particles show poor flowability at the pilot scale, while the presence of fine materials (less than 0.7 mm) and bark improves the flowability of biomass slurries without affecting the organic acid yields (based on C6 carbohydrate content).

09 - BIOMASS FUELS↗

DOE OSTI2021

Severity factor kinetic model as a strategic parameter of hydrothermal processing (steam explosion and liquid hot water) for biomass fractionation under biorefinery concept

Hydrothermal processes are an attractive clean technology and cost-effective engineering platform for bio-refineries based in the conversion of biomass to biofuels and high-value bioproducts under the basis of sustainability and circular bioeconomy. The deep and detailed knowledge of the structural changes by the severity of biomasses hydrothermal fractionation is scientifically and technological needed in order to improve processes effectiveness, reactors designs, and industrial application of the multi-scale target compounds obtained by stream explosion and liquid hot water systems. The concept of the severity factor [log 10 (R 0 )] established > 30 years ago, continue to be a useful index that can provide a simple description of the relationship between the operational conditions for biomass fractionation in second generation of biorefineries. Furthermore, this review develops a deep explanation of the hydrothermal severity factor based in lignocellulosic biomass fractionation with emphasis in research advances, pretreatment operations and the applications of severity factor kinetic model.

09 BIOMASS FUELS↗

DOE OSTI2022

Utilization of guaiacol-based deep eutectic solvent for achieving a sustainable biorefinery

This study proposed a renewable deep eutectic solvent (DES) pretreatment using lignin-derived guaiacol as the hydrogen bond donor. The DES showed excellent biomass fractionation efficiency after the incorporation of trace AlCl 3 as the reinforcer, which removed 79.1 % lignin while preserving more than 90 % glucan. The pretreated bamboo exhibited 96.2 % glucan enzymatic hydrolysis yield at only 110 °C. The physicochemical properties of the pretreated solids were comprehensively investigated to explain how the DES fractionation overcame the biomass recalcitrance. The regenerated lignin from the DES pretreatment was also analyzed, which revealed that lignin β-O-4 bond was significantly cleaved. In conclusion, this guaiacol-based DES could greatly contribute to establish a closed-loop biorefinery sequence with high lignin fractionation efficiency and great solvent recyclability.

09 BIOMASS FUELS↗

DOE OSTI2025

Redefining the product portfolio of oilcane bagasse biorefinery: Recovering natural colorants, vegetative lipids and sugars

Bioenergy crops have been known for their ability to produce biofuels and bioproducts. In this study, the product portfolio of recently developed transgenic sugarcane (oilcane) bagasse has been redefined for recovering natural pigments (anthocyanins), sugars, and vegetative lipids. The total anthocyanin content in oilcane bagasse has been estimated as 92.9 ± 18.9 µg/g of dried bagasse with cyanidin-3-glucoside (13.5 ± 18.9 µg per g of dried bagasse) as the most prominent anthocyanin present. More than 85 % (w/w) of the total anthocyanins were recovered from oilcane bagasse at a pretreatment temperature of 150 °C for 15 min. These conditions for the hydrothermal pretreatment also led to a 2-fold increase in the glucose yield upon the enzymatic saccharification of the pretreated bagasse. Further, a 1.5-fold enrichment of the vegetative lipids was demonstrated in the pretreated residue. Re-defining green biorefineries with multiple high-value products in a zero-waste approach is the need of the hour for attaining sustainability.

09 BIOMASS FUELS↗

DOE OSTI2022

Advances and perspectives on mass transfer and enzymatic hydrolysis in the enzyme-mediated lignocellulosic biorefinery: A review

We report enzymatic hydrolysis is a critical process for the cellulase-mediated lignocellulosic biorefinery to produce sugar syrups that can be converted into a whole range of biofuels and biochemicals. Such a process operating at high-solid loadings (i.e., scarcely any free water or roughly ≥ 15% solids, w/w) is considered more economically feasible, as it can generate a high sugar concentration at low operation and capital costs. However, this approach remains restricted and incurs “high-solid effects”, ultimately causing the lower hydrolysis yields with increasing solid loadings. The lack of available water leads to a highly viscous system with impaired mixing that exhibits strong transfer resistance and reaction limitation imposed on enzyme action. Evidently, high-solid enzymatic hydrolysis involves multi-scale mass transfer and multi-phase enzyme reaction, and thus requires a synergistic perspective of transfer and biotransformation to assess the interactions among water, biomass components, and cellulase enzymes. Porous particle characteristics of biomass and its interface properties determine the water form and distribution state surrounding the particles, which are summarized in this review aiming to identify the water-driven multi-scale/multi-phase bioprocesses. Further aided by the cognition of rheological behavior of biomass slurry, solute transfer theories, and enzyme kinetics, the coupling effects of flow-transfer-reaction are revealed under high-solid conditions. Based on the above basic features, this review lucidly explains the causes of high-solid hydrolysis hindrances, highlights the mismatched issues between transfer and reaction, and more importantly, presents the advanced strategies for transfer and reaction enhancements from the viewpoint of process optimization, reactor design, as well as enzyme/auxiliary additive customization.

45 multi-scale, multi-phase, enzymatic reaction↗

DOE OSTI2022

Biocatalytic gateway to convert glycerol into 3-hydroxypropionic acid in waste-based biorefineries: Fundamentals, limitations, and potential research strategies

Microbial conversion of bioenergy-derived waste glycerol into value-added chemicals has emerged as an important bioprocessing technology due to its eco-friendliness, feasible technoeconomics, and potential to provide sustainability in biodiesel and bioethanol production. Glycerol is an abundant liquid waste from bioenergy plants with a projected volume of 6 million tons by 2025, accounting for about 10% of biodiesel and 2.5% of bioethanol yields. 3-Hydroxypropionic acid (3-HP) is a major product of glycerol bioconversion, which is the third largest biobased platform compound with expected market size and value of 3.6 million tons/year and USD 10 billion/year, respectively. Despite these biorefinery values, 3-HP biosynthesis from glycerol is still at an immature stage of commercial exploitation. The main challenges behind this immaturity are the toxic effects of 3-HPA on cells, the distribution of carbon flux to undesirable pathways, low tolerance of cells to glycerol and 3-HP, co-factor dependence of enzymes, low enzyme activity and stability, and the problems of substrate inhibition and specificity of enzymes. To address these challenges, it is necessary to understand the fundamentals of glycerol bioconversion and 3-HP production in terms of metabolic pathways, related enzymes, cell factories, midstream process configurations, and downstream 3-HP recovery, as discussed in this review critically and comprehensively. It is equally important to know the current challenges and limitations in 3-HP production, which are discussed in detail along with recent research efforts and remaining gaps. Finally, possible research strategies are outlined considering the recent technological advances in microbial biosynthesis, aiming to attract further research efforts to achieve a sustainable and industrially exploitable 3-HP production technology. Here, by discussing the use of advanced tools and strategies to overcome the existing challenges in 3-HP biosynthesis, this review will attract researchers from many other similar biosynthesis technologies and provide a common gateway for their further development.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2026

Green pretreatment strategies for enhanced microbial lipid fermentation and synergistic high-quality lignin recovery for next-generation integrated biorefineries

Miscanthus × giganteus (Mxg) is a warm-season perennial grass being commercialized as a biomass feedstock for temperate farms. Three process strategies were compared for converting Mxg into single-cell oil and extracted lignin intermediates, as hydrothermal (HT) processing and two natural deep eutectic solvents (NADES), ChCl:LA (choline chloride:lactic acid) and ChCl:Gly (choline chloride:glycerol). Pretreatments were performed at 10% and 50% solids loading at 140 °C, 2 h; HT: 190 °C, 10 min. Enzymatic hydrolysates, generated at 10% solids using washed and unwashed biomass, were evaluated for microbial lipid production. Maximum glucose conversions from washed biomass reached 83.5% (ChCl:LA), 52.7% (ChCl:Gly), and 74.0% (HT). Remarkably, NADES-derived hydrolysates effectively replaced refined sugars for the cultivation of the oleaginous yeast Rhodotorula toruloides, achieving ∼51% higher biomass (OD 90.7) and lipid titers of 19.36 g/L within 45 h using a two-stage fermentation strategy. Lipid contents ranged from 34.5–44.5% dry weight, demonstrating reduced reliance on purified sugars. Beyond carbohydrate valorization, ChCl:LA pretreatment enabled high-purity lignin recovery (>89%) in a lignin-first strategy. Structural analyses (2D-HSQC and ³¹P NMR) showed syringyl (78.15%), guaiacyl (15.15%), and p-hydroxyphenyl (6.41%) units, with higher phenolic hydroxyl content (0.91 mmol/g) and a lower S/G ratio (0.19) than HT lignin (0.87 mmol/g, S/G 0.22). These attributes favor downstream lignin depolymerization into low-molecular-weight aromatics. Overall, NADES pretreatment simultaneously enhances microbial lipid yields and recovers high-quality lignin, advancing the economic feasibility of renewable diesel and sustainable aviation fuel (SAF) production from bioenergy crops within an integrated biorefinery framework.

2D HSQC↗

DOE OSTI2026

Evaluating CO 2 mitigation strategies in SAF biorefineries: Techno-economic and life cycle analysis

The aviation sector requires scalable decarbonization strategies, and lignocellulosic sustainable aviation fuel (SAF) represents a promising pathway. This study comparatively evaluates the techno-economic analysis and life cycle assessment (LCA) of three CO 2 management strategies integrated within a U.S.-based gasification–Fischer–Tropsch SAF biorefinery: (i) catalytic hydrogenation of captured CO 2 to methanol, (ii) geological CO 2 sequestration, and (iii) mineralization to sodium bicarbonate (NaHCO 3 ). Techno-economic analysis indicates that methanol synthesis requires approximately 26% higher capital investment and 33% higher operating costs than mineralization. Although methanol co-production generates the highest gross revenue, NaHCO 3 production reduces the SAF minimum selling price by approximately 38% relative to both methanol synthesis and geological sequestration pathways, reflecting a more balanced cost allocation through mineral co-product valorization. Geological sequestration lowers operating costs by nearly 50% compared with methanol synthesis but remains highly dependent on carbon credit mechanisms. LCA reveals substantial divergence in climate performance. Relative to methanol synthesis, sequestration improves net greenhouse gas performance by approximately 163%, transitioning the system from net-positive to net-negative emissions. Mineralization further enhances carbon mitigation, achieving roughly 85% greater carbon reduction than sequestration and over sixfold improvement relative to methanol synthesis within the defined system boundary. Sensitivity analysis identified hydrogen price, co-product market value, and process emissions as dominant drivers. Under baseline assumptions, CO 2 mineralization is found to offer the most balanced pathway.

Carbon capture and storage↗

DOE OSTI2020

Life cycle greenhouse gas emissions of different CO 2 supply options for an algal biorefinery

This paper characterizes the greenhouse gas (GHG) emissions from the provision of CO 2 for twelve hours of the day from a range of sources operating on a twenty-four-hour basis. Twelve CO 2 supply cases spanning from fossil fuel power plant stack gases to direct air capture (DAC) systems are modeled. The results are compared in a life cycle assessment (LCA) framework. The CO 2 supply from the combustion or gasification of biomass has GHG emissions of –1.62 gCO 2 eq./g CO 2 delivered and is the case with the lowest carbon footprint for supplied CO2. The emissions for all the CO 2 supply cases were found to be negative, either because of capturing more CO 2 from the atmosphere than is released from the CO 2 production process, or because of production of excess electricity that is exported to the grid (replacing emissions from grid electricity). A purpose-built Natural Gas Combined Cycle (NGCC) power plant CO 2 source has the highest GHG emissions of -0.40 gCO 2 eq./gCO 2 delivered of the cases considered because of nighttime emissions and natural gas supply chain losses. The impact of a diurnal cycle can be mitigated by using capture and refrigeration systems as part of the CO 2 management system. The results are sensitive to the GHG emissions of the grid electricity, which is imported or exported in different cases. The results provide a benchmark for the comparison of different CO 2 supply options for the establishment of a sustainable algal biorefinery.

42 ENGINEERING↗

DOE OSTI2024

Valorization of lignin from aqueous-based lignocellulosic biorefineries

An additional 100 million tons/year of lignin coproduct will result when lignocellulosic biomass is processed in biorefineries to fiber, sugars, biofuels, and bioproducts. This will double the amount of lignin already generated from pulping and paper production. Unlike pulping that results in lignosulphonate (88% of total) or Kraft lignin (9%), aqueous-based biorefining leaves behind non-sulfonated lignin and aromatic molecules. Furthermore, this new type of lignin provides opportunities for large volume agricultural uses such as controlled-release carriers and soil amendments as well as feedstocks for new chemistries that lead to molecular building blocks for the chemical industry and to precursors for sustainable aviation biofuels.

03 NATURAL GAS↗

DOE OSTI2026

A Catabolic Powerhouse for Biorefineries: Characterization and Engineering Erwinia spp. Strain LJJL01 to Produce Bioproducts

Nonmodel microbial hosts can efficiently biotransform unconventional organic feedstocks into advanced bioproducts, leveraging their superior metabolic capabilities and resilience to process-relevant physicochemical conditions. In this study, we domesticated Erwinia spp. strain. LJJL01 (Er LJJL01) as a potent microbial chassis to advance the emerging biorefinery strategy for the valorization of lignin-rich biomass and plastic, thereby enabling the circular economy. The strain exhibits remarkable chemical tolerance and can biofunnel various substrates, including sugars, acids, polyols, and aromatics, in minimal salt media to produce native fine chemicals such as acetoin, 2,3-butanediol, and lactic acid. As a proof of concept, engineering this strain with advanced genetic tools enables the production of tailored high-value chemicals, such as cis,cis-muconate from plastic-derived terephthalate and polyhydroxybutyrate from lignocellulosic hydrolysate. We established Er LJJL01 as a potent microbial chassis for the green synthesis of bioproducts from unconventional organic feedstocks.

09 BIOMASS FUELS↗

DOE OSTI2020

Materials for the biorefinery: high bio-content, shape memory Kraft lignin-derived non-isocyanate polyurethane foams using a non-toxic protocol

Polyurethanes are among the top six polymers produced in the world and are widely used in the automotive, furniture, construction and appliance industry for their light weight, impact resistance, and insulating properties. However, the use of hazardous diisocyanates used in polyurethane formulations has led many to search for more sustainable alternatives. The lignin component of biomass has been targeted to replace the often toxic and petroleum-derived precursors to polymer synthesis in support of the biorefinery concept where natural materials are used as the feedstock for commodity plastics. The use of lignin is often hampered due to its low reactivity, heterogeneity and the necessity of employing extensive purification and/or functionalization measures to ensure materials are of comparable quality. In this report, a unique method is presented for the synthesis and processing of a non-toxic, non-isocyanate polyurethane foam utilizing unmodified Kraft lignin and a biobased curing agent. Here, raw Kraft lignin is functionalized with green organic carbonates and a non-toxic approach is taken to solubilize the precursors with a curing agent from renewable fatty acids enabling rapid gel-times. For the first time, a NIPU foam from lignin is synthesized while the structure–property relationship of different reaction mixtures is studied demonstrating shape memory capacity and 100% biobased carbon content.

36 MATERIALS SCIENCE↗