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At least 145 records · Page 8

Life-Cycle Assessment of Sustainable Aviation Fuel Derived from Paper Sludge

Converting waste paper sludge to sustainable aviation fuel (SAF) offers a circular economy strategy to decarbonize the aviation sector. Here, this study develops a life-cycle assessment (LCA) for converting high-ash paper sludge to SAF in the U.S. using a catalytic sugar upgrading system that consists of ash removal, enzymatic hydrolysis, dehydration, aldol condensation, and hydroprocessing. The LCA is coupled with a process simulation for an industrial-scale biorefinery based on experimental data. We quantified the carbon intensity as 35.7–41.8 gCO 2 eq MJ –1 SAF (–636 to –584 gCO 2 eq per dry kg paper sludge) with acetone as a solvent, renewable fuel, and biobased chemicals; this is further reduced to 5.1–11.1 gCO 2 eq MJ –1 (–925 to –873 gCO 2 eq per dry kg paper sludge) if ash is recycled and used for substituting cement. Converting 1 dry kg paper sludge to SAF with acetone, renewable fuel, and biobased chemicals (–925 to –584 gCO 2 eq) is more climate beneficial than landfilling without landfill gas recovery (791 gCO 2 eq) and with landfill gas recovery (–294 gCO 2 eq). More than 330 million gallons of SAF can be produced annually (>4 million dry t paper sludge/year in the U.S.), resulting in a reduction of 2–7 million tCO 2 eq.

09 BIOMASS FUELS↗

Preserving Aryl Ether Linkages and Higher Yields of Isolated Lignin through Biomass Fibrillation

Ball milling is used for size reduction of biomass for isolating lignin. However, it is known that it can lead to some structural alteration of lignin. Fibrillation using ultrafriction grinding is a newer size reduction technology which was applied to poplar for isolating cellulolytic enzyme lignin. Compositional analysis of poplar, enzymatic hydrolysis yield, water retention value, lignin yield, lignin structural characterization, and lignin molecular weight analysis were carried out to compare lignin isolated from fibrillated and ball milled poplar. By using fibrillation instead of planetary ball milling for size reduction of poplar, higher β-O-4' linkage content, lignin yield, and molecular weight were obtained. Replacing ball milling with fibrillation for size reduction allows recovery of lignin whose structure is closer to that of lignin in biomass.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Tailoring poplar lignin without yield penalty by combining a null and haploinsufficient CINNAMOYL-CoA REDUCTASE2 allele

Lignin causes lignocellulosic biomass recalcitrance to enzymatic hydrolysis. Engineered low-lignin plants have reduced recalcitrance but often exhibit yield penalties, offsetting their gains in fermentable sugar yield. Here, CRISPR/Cas9-generated CCR2(–/*) line 12 poplars have one knockout CCR2 allele while the other contains a 3-bp deletion, resulting in a 114I115A-to-114T conversion in the corresponding protein. Despite having 10% less lignin, CCR2(–/*) line 12 grows normally. On a plant basis, the saccharification efficiency of CCR2(–/*) line 12 is increased by 25–41%, depending on the pretreatment. Analysis of monoallelic CCR2 knockout lines shows that the reduced lignin amount in CCR2(–/*) line 12 is due to the combination of a null and the specific haploinsufficient CCR2 allele. Analysis of another CCR2(–/*) line shows that depending on the specific CCR2 amino-acid change, lignin amount and growth can be affected to different extents. Furthermore, our findings open up new possibilities for stably fine-tuning residual gene function in planta.

59 BASIC BIOLOGICAL SCIENCES↗

Carbohydrate-aromatic interface and molecular architecture of lignocellulose

Abstract Plant cell walls constitute the majority of lignocellulosic biomass and serve as a renewable resource of biomaterials and biofuel. Extensive interactions between polysaccharides and the aromatic polymer lignin make lignocellulose recalcitrant to enzymatic hydrolysis, but this polymer network remains poorly understood. Here we interrogate the nanoscale assembly of lignocellulosic components in plant stems using solid-state nuclear magnetic resonance and dynamic nuclear polarization approaches. We show that the extent of glycan-aromatic association increases sequentially across grasses, hardwoods, and softwoods. Lignin principally packs with the xylan in a non-flat conformation via non-covalent interactions and partially binds the junction of flat-ribbon xylan and cellulose surface as a secondary site. All molecules are homogeneously mixed in softwoods; this unique feature enables water retention even around the hydrophobic aromatics. These findings unveil the principles of polymer interactions underlying the heterogeneous architecture of lignocellulose, which may guide the rational design of more digestible plants and more efficient biomass-conversion pathways.

59 BASIC BIOLOGICAL SCIENCES↗

A biomass pretreatment using cellulose-derived solvent Cyrene

Despite only recently becoming available in the quantities required for solvent usage, the cellulose-derived solvent, named Cyrene, has gained significant attention in green chemistry in recent years. To fulfill the sustainability criteria of future biorefineries, a novel renewable biomass pretreatment using Cyrene and water was developed for the first time. Results showed that Cyrene has high potential as a green pretreatment solvent in terms of lignin fractionation/recovery and sugar release in the follow-up enzymatic hydrolysis. The mechanism of this pretreatment was revealed by investigating the structural characteristics of pretreated biomass, and the recovered lignin was also fully characterized to assess its valorization potential. Results indicated that Cyrene pretreatment could be performed at a mild condition (120 °C) to reduce the lignin condensation and the cleavage of β-O-4 linkages without compromising lignin removal and the following sugar platform. The successful utilization of this cellulose-derived solvent in pretreatment will further contribute to the realization of a “closed-loop” biorefinery process.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Development of an ammonia pretreatment that creates synergies between biorefineries and advanced biomass logistics models

In this study, a novel ammonia-based pretreatment for densified lignocellulosic biomass was developed to reduce ammonia usage and integrate with viable biomass logistics scenarios. The COmpacted Biomass with Recycled Ammonia (COBRA) pretreatment performed at 100 °C allows >95% conversion of sugarcane bagasse (SCB) carbohydrates into soluble monomeric and oligomeric sugars (glucose and xylose) using industrially relevant 6% glucan loading (~21% solids loading) enzymatic hydrolysis conditions at reduced enzyme loadings. Pretreatment via COBRA with simultaneous lignin extraction (COBRA-LE) improved Saccharomyces cerevisiae 424A(LNH-ST) metabolic yield from 89% to 97.5% relative to COBRA without delignification, allowing a process ethanol yield of 71.6%. A technoeconomic analysis on SCB biorefining to ethanol in the state of São Paulo, Brazil, compared COBRA to other mature technologies, such as AFEX and steam-explosion. Amongst all scenarios studied, biorefineries based on COBRA-LE pretreatment offered the lowest average minimum ethanol selling price of US$1.45 per gallon ethanol. COBRA pretreatment was subsequently tested on perennial grasses and hardwoods, and >80% total sugar yields were achieved for all cases.

09 BIOMASS FUELS↗

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↗

Demonstrating a butylamine-based deconstruction method for poplar biomass and conversion by diverse microbial strains

Low-boiling alkylamines such as butylamine offer promise as effective biomass pretreatment solvents that can be readily recovered and recycled; however, their capability to support microbial conversion of nutrients present in hydrolysates represents an important area for investigation. Here we employed butylamine to pretreat poplar biomass and characterize its effects on the release of fermentable sugars after solvent removal and enzymatic hydrolysis, as well as the biocompatibility of the produced hydrolysates with three organisms commonly used as bioconversion hosts. We observed that residual butylamine and the derivative butylacetamide were present in high enough concentrations to exert toxicity to strains of Aspergillus niger, Pseudomonas putida, and Rhodosporidium toruloides that produce malic acid, isoprenol and bisabolene, respectively. Removal of the toxic compounds by charcoal filtration and nutrient supplementation resulted in a hydrolysate containing >100 g L −1 of sugars that enabled strong growth, substrate consumption and bioproduct accumulation, outperforming defined cultivation media. This is the first demonstration of a butylamine-based deconstruction process for poplar biomass at a pilot-scale to achieve conversion of high sugar concentrations to valuable bioproducts with engineered microbes.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Feedstock variability impacts the bioconversion of sugar and lignin streams derived from corn stover by Clostridium tyrobutyricum and engineered Pseudomonas putida

Abstract Feedstock variability represents a challenge in lignocellulosic biorefineries, as it can influence both lignocellulose deconstruction and microbial conversion processes for biofuels and biochemicals production. The impact of feedstock variability on microbial performance remains underexplored, and predictive tools for microbial behaviour are needed to mitigate risks in biorefinery scale‐up. Here, twelve batches of corn stover were deconstructed via deacetylation, mechanical refining, and enzymatic hydrolysis to generate lignin‐rich and sugar streams. These batches and their derived streams were characterised to identify their chemical components, and the streams were used as substrates for producing muconate and butyrate by engineered Pseudomonas putida and wildtype Clostridium tyrobutyricum , respectively. Bacterial performance (growth, product titers, yields, and productivities) differed among the batches, but no strong correlations were identified between feedstock composition and performance. To provide metabolic insights into the origin of these differences, we evaluated the effect of twenty‐three isolated chemical components on these microbes, including three components in relevant bioprocess settings in bioreactors, and we found that growth‐inhibitory concentrations were outside the ranges observed in the streams. Overall, this study generates a foundational dataset on P. putida and C. tyrobutyricum performance to enable future predictive models and underscores their resilience in effectively converting fluctuating lignocellulose‐derived streams into bioproducts.

09 BIOMASS FUELS↗

Enzyme kinetics by GH7 cellobiohydrolases on chromogenic substrates is dictated by non‐productive binding: insights from crystal structures and MD simulation

Cellobiohydrolases (CBHs) in the glycoside hydrolase family 7 (GH7) ( EC3.2.1.176 ) are the major cellulose degrading enzymes both in industrial settings and in the context of carbon cycling in nature. Small carbohydrate conjugates such as p‐ nitrophenyl‐β‐ d ‐cellobioside (pNPC), p‐ nitrophenyl‐β‐ d ‐lactoside (pNPL) and methylumbelliferyl‐β‐ d ‐cellobioside have commonly been used in colorimetric and fluorometric assays for analysing activity of these enzymes. Despite the similar nature of these compounds the kinetics of their enzymatic hydrolysis vary greatly between the different compounds as well as among different enzymes within the GH7 family. Through enzyme kinetics, crystallographic structure determination, molecular dynamics simulations, and fluorometric binding studies using the closely related compound o‐nitrophenyl‐β‐ d ‐cellobioside (oNPC), in this work we examine the different hydrolysis characteristics of these compounds on two model enzymes of this class, TrCel7A from Trichoderma reesei and PcCel7D from Phanerochaete chrysosporium . Protein crystal structures of the E212Q mutant of TrCel7A with pNPC and pNPL, and the wildtype TrCel7A with oNPC, reveal that non‐productive binding at the product site is the dominating binding mode for these compounds. Enzyme kinetics results suggest the strength of non‐productive binding is a key determinant for the activity characteristics on these substrates, with PcCel7D consistently showing higher turnover rates ( k cat ) than TrCel7A, but higher Michaelis–Menten ( K M ) constants as well. Furthermore, oNPC turned out to be useful as an active‐site probe for fluorometric determination of the dissociation constant for cellobiose on TrCel7A but could not be utilized for the same purpose on PcCel7D, likely due to strong binding to an unknown site outside the active site.

59 BASIC BIOLOGICAL SCIENCES↗

Virtual Engineering (VE) (vebio)

The Virtual Engineering (VE, vebio) project provides a set of Python modules to enable users to connect mathematical models of unit operations to predict outcomes for an entire chemical process. This VE approach is currently being developed to support the beginning-to-end simulation of the low-temperature conversion of lignocellulosic biomass to a fuel precursor, but the underlying communication functions can be readily extended to connect models from other domains. The models and features currently included in the VE library include a models/simulations for pretreatment, enzymatic hydrolysis, and aerobic bioreaction.

Young, Ethan↗

Water-soluble saponins accumulate in drought-stressed switchgrass and may inhibit yeast growth during bioethanol production

Abstract Background Developing economically viable pathways to produce renewable energy has become an important research theme in recent years. Lignocellulosic biomass is a promising feedstock that can be converted into second-generation biofuels and bioproducts. Global warming has adversely affected climate change causing many environmental changes that have impacted earth surface temperature and rainfall patterns. Recent research has shown that environmental growth conditions altered the composition of drought-stressed switchgrass and directly influenced the extent of biomass conversion to fuels by completely inhibiting yeast growth during fermentation. Our goal in this project was to find a way to overcome the microbial inhibition and characterize specific compounds that led to this inhibition. Additionally, we also determined if these microbial inhibitors were plant-generated compounds, by-products of the pretreatment process, or a combination of both. Results Switchgrass harvested in drought (2012) and non-drought (2010) years were pretreated using Ammonia Fiber Expansion (AFEX). Untreated and AFEX processed samples were then extracted using solvents (i.e., water, ethanol, and ethyl acetate) to selectively remove potential inhibitory compounds and determine whether pretreatment affects the inhibition. High solids loading enzymatic hydrolysis was performed on all samples, followed by fermentation using engineered Saccharomyces cerevisiae . Fermentation rate, cell growth, sugar consumption, and ethanol production were used to evaluate fermentation performance. We found that water extraction of drought-year switchgrass before AFEX pretreatment reduced the inhibition of yeast fermentation. The extracts were analyzed using liquid chromatography–mass spectrometry (LC–MS) to detect compounds enriched in the extracted fractions. Saponins, a class of plant-generated triterpene or steroidal glycosides, were found to be significantly more abundant in the water extracts from drought-year (inhibitory) switchgrass. The inhibitory nature of the saponins in switchgrass hydrolysate was validated by spiking commercially available saponin standard (protodioscin) in non-inhibitory switchgrass hydrolysate harvested in normal year. Conclusions Adding a water extraction step prior to AFEX-pretreatment of drought-stressed switchgrass effectively overcame inhibition of yeast growth during bioethanol production. Saponins appear to be generated by the plant as a response to drought as they were significantly more abundant in the drought-stressed switchgrass water extracts and may contribute toward yeast inhibition in drought-stressed switchgrass hydrolysates.

59 BASIC BIOLOGICAL SCIENCES↗

Data for Hydrothermal Pretreatment for Valorization of Genetically Engineered Bioenergy Crop for Lipid and Cellulosic Sugar Recovery

Lipids accumulated in the vegetative tissues of cellulosic feedstocks can be a potential raw material for biodiesel and bioethanol production. In this work, bagasse of genetically engineered sorghum was subjected to liquid hot-water pretreatment at 170, 180, and 190 °C for different reaction time. Under the optimal pretreatment condition (170 °C, 20 min), the residue was enriched in glucan (57.39 ± 2.63 % w/w) and xylan (13.38 ± 0.49 % w/w). The total lipid content of the pretreated residue was 6.81% w/w, similar to that observed in untreated bagasse (6.30% w/w). Pretreatment improved the enzymatic digestibility of bagasse, allowing a recovery of 79% w/w and 86% w/w of glucose and xylose, respectively. The pretreatment and enzymatic saccharification resulted in a 2-fold increase in total lipid in enzymatic residue compared to the original bagasse. Thus, pretreatment and enzymatic hydrolysis enabled high sugar recovery while concentrating triglycerides and free fatty acids in the residue.

Conversion↗

Data for Natural Deep Eutectic Solvents (NADES) Assisted Deconstruction of Oilcane Bagasse for High Lipid and Sugar Recovery

In a novel approach, metabolically engineered sugarcane “Oilcane” has been investigated for fractionation of lipid and cellulose-rich pulp, using certain Natural deep eutectic solvents (NADES). The exploration of eco-friendly solvents are at the forefront of harnessing the biofuel potential of modern bioenergy crops. For this, six combinations of NADES were prepared using choline chloride (ChCl) as HBA and lactic acid (LA), oxalic acid (OA) and glycerol (Gly) as HBD and were further explored for pretreatment of oilcane bagasse in a molar ratio of 1:1 and 1:2. The impact of NADES ratio, biomass loading (10–50%), residence time (1–2 h), and temperature (90–140 °C) were evaluated for delignification, lipid content, sugar release after enzymatic hydrolysis. The finding demonstrated that under the optimal condition of ChCl: LA (1:2 molar ratio), 140 °C with 2 h retention time, the lipid content in the pre-treated substrate was increased to 2.5-fold (∼8% w/w) and > 80% glucose yield was achieved after 72 h of hydrolysis of pre-treated bagasse. High solid loading (∼50%) during pretreatment resulted in a similar glucose yield. Furthermore, recycling studies demonstrated that nearly 95 to 98% NADES could be recycled after each pretreatment for up to five consecutive cycles without any significant loss in chemical structure as confirmed by 1H NMR and FT IR. FT IR and XRD analyses of native and pre-treated biomass were performed to visualize the morphological changes during NADES pretreatment and their impact on sugar yield. The findings of the study may be used to establish NADES-based biorefinery for the valorization of lipids, and carbohydrates for fuels and chemicals production.

Conversion↗

Data for Valorization of Miscanthus x giganteus for Sustainable Recovery of Anthocyanins and Enhanced Production of Sugars

The increased awareness for eco-friendliness and sustainability has shifted the interest of stakeholders from synthetic colors to natural plant-based pigments. In this study, purple stemmed Miscanthus x giganteus was evaluated as a source of anthocyanins. Hydrothermal pretreatment was studied as a green, chemical-free process for recovering maximum anthocyanins in the pretreatment liquor. The highest recovery of 94.3 ± 1.5% w/w of the total anthocyanin concentration was obtained for a temperature and time combination of 170 °C and 10 min. The pretreatment also improved the enzymatic digestibility of the biomass and led to a 2.1-fold increase in the overall recovery of glucose (70.6 ± 0.5% w/w) at the end of 72 h. The sugar monomers obtained after the enzymatic hydrolysis of the pretreated biomass could be used for the production of biofuels or biochemicals in an integrated biorefinery based on purple-stemmed miscanthus. Overall, this study demonstrates that the clean pretreatment method developed could lead to an additional product stream (rich in anthocyanins) along with its effect in reducing the recalcitrance of miscanthus biomass.

Biomass Analytics↗

Data for Production of Designer Xylose-Acetic Acid Enriched Hydrolysate from Bioenergy Sorghum, Oilcane, and Energycane Bagasses

Xylan accounts for up to 40% of the structural carbohydrates in lignocellulosic feedstocks. Along with xylan, acetic acid in sources of hemicellulose can be recovered and marketed as a commodity chemical. Through vibrant bioprocessing innovations, converting xylose and acetic acid into high-value bioproducts via microbial cultures improves the feasibility of lignocellulosic biorefineries. Enzymatic hydrolysis using xylanase supplemented with acetylxylan esterase (AXE) was applied to prepare xylose-acetic acid enriched hydrolysates from bioenergy sorghum, oilcane, or energycane using sequential hydrothermal-mechanical pretreatment. Various biomass solids contents (15 to 25%, w/v) and xylanase loadings (140 to 280 FXU/g biomass) were tested to maximize xylose and acetic acid titers. The xylose and acetic acid yields were significantly improved by supplementing with AXE. The optimal yields of xylose and acetic acid were 92.29% and 62.26% obtained from hydrolyzing energycane and oilcane at 25% and 15% w/v biomass solids using 280 FXU xylanase/g biomass and AXE, respectively.

Biomass Analytics↗

Techno-Economic Case Study: Low-Temperature Conversion Performance Based on Isolated Anatomical Fractions of Corn Stover

This report summarizes analysis conducted to support a case study under the Feedstock-Conversion Interface Consortium (FCIC) focused on techno-economic analysis (TEA) modeling to quantify the yield and cost ramifications for processing isolated anatomical fractions of corn stover through a low-temperature conversion (biochemical pathway) biorefinery. It is hypothesized that different individual anatomical fractions of corn stover vary in composition and recalcitrance, such that processing each fraction on its own e.g. through dedicated campaigns may enable economic benefits relative to processing the whole stover material. To investigate this concept, we worked with FCIC researchers to obtain data on compositional analysis, as well as processing conditions and yields, for conversion of whole corn stover plus three individual anatomical fractions (cobs, husks, and stalks) across key steps of the biorefinery conversion process within FCIC research scope, namely pretreatment and enzymatic hydrolysis. After running the data through the TEA models, fractionated cobs and husks achieved higher fuel yields and lower minimum fuel selling prices than whole corn stover, but stalks fared worse than whole stover on both metrics. The most direct takeaway from this assessment is that a biorefinery would stand to benefit economically from maximizing the use of cob and husk fractions while avoiding or minimizing the use of stalks, though recognizing practical constraints with this approach in maintaining an equivalent processing capacity (fixed at 2,000 dry metric tonne per day in all cases).

09 BIOMASS FUELS↗

How does pelleting alter the enzymatic digestibility of corn stover?

Pelleting of lignocellulosic biomass facilitates its transportation, storage and handling and offers opportunities to improve the enzymatic digestibility of pelleted biomass. Here, we report the impact of pelleting on the enzymatic digestibility of corn stover prior to pretreatment and associated substrate characteristics. Pelleting almost doubles the digestibility of unpretreated corn stover, from 8.2 to 15.5% glucan conversion, at 5% solid loading using 1 FPU Cellic CTec2 per g solids. Compositional analysis indicates that loose and pelleted corn stover have similar non-dissolvable compositions, although their extractives are slightly different. Enzymatic hydrolysis of washed corn stover after size reduction to normalize particle sizes and removal of extractives confirms that pelleting improves corn stover digestibility. Such differences may be explained by the decreased particle size, increased specific surface area, improved cellulose accessibility, and reduction of the polymer length of cellulose induced by pelleting. These findings help to design processing schemes for sustainable and efficient use of lignocellulose.

Chen, Xueli↗