Illinois Data Bank Dataset Search Results
Results
published:
2025-11-03
Banerjee, Shivali; Dien, Bruce; Eilts, Kristen; Sacks, Erik; Singh, Vijay
(2025)
Chemical-free hydrothermal pretreatment of Miscanthus x giganteus (Mxg) at the lab scale using high liquid-to-solid ratios resulted in the recovery of anthocyanins and enhanced enzymatic digestibility of residual biomass. In this study, the process is scaled up by using a continuous hydrothermal pretreatment reactor operated at a low liquid-to-solid ratio (50 % w/w solids) as an important step towards commercialization. Anthocyanin yield was 70 % w/w at the pilot scale (50 kg of Mxg), compared to the 94 % w/w yield achieved at the lab scale (0.5 g of Mxg). The pretreated biomass was subsequently refined mechanically using a disc mill to increase the accessibility of cellulose by cellulases. Enzymatic saccharification of the pretreated and disc-milled residue yielded 238 g/L sugar concentration by operating in fed-batch mode at 50 % w/v solids content. Two strains of Rhodosporidium toruloides were evaluated for converting the hydrolysate sugars into microbial lipids, and strain Y-6987 had the highest lipid titer (11.0 g/L). Further, the residue left after enzymatic saccharification was determined to be enriched 1.7-fold in the lignin content. This lignin-rich residue has value as a feedstock for the production of sustainable aviation fuel precursors and other high-value lignin-based chemicals. Hence the proposed biorefinery based on Mxg creates an opportunity for generating revenue from multiple high-value products. As the demand for biofuels and biobased products is rising, the biorefinery products from Mxg would create a niche in the industrial sector.
keywords:
Conversion;Feedstock Production;Feedstock Bioprocessing;Hydrolysate;Lipidomics
published:
2025-11-03
Anaokar, Sanket; Liang, Yuanxue; Yu, Xiao-Hong; Cai, Yingqi; Cai, Yuanheng; Shanklin, John
(2025)
Triacylglycerols (TAG), accumulate within lipid droplets (LD), predominantly surrounded by OLEOSINs (OLE), that protect TAG from hydrolysis. We tested the hypothesis that identifying and removing degradation signals from OLE would promote its abundance, preventing TAG degradation and enhancing TAG accumulation. We tested whether mutating potential ubiquitin-conjugation sites in a previously reported improved Sesamum indicum OLE (SiO) variant, o3-3 Cys-OLE (SiCO herein), would stabilize it and increase its lipogenic potential. SiCOv1 was created by replacing all five lysines in SiCO with arginines. Separately, six cysteine residues within SiCO were deleted to create SiCOv2. SiCOv1 and SiCOv2 mutations were combined to create SiCOv3. Transient expression of SiCOv3 in Nicotiana benthamiana increased TAG by two-fold relative to SiCO. Constitutive expression of SiCOv3 or SiCOv5, containing the five predominant TAG-increasing mutations from SiCOv3, in Arabidopsis along with mouse DGAT2 (mD) increased TAG accumulation by 54% in leaves and 13% in seeds compared with control lines coexpressing SiCO and mD. Lipid synthesis rates increased, consistent with an increase in lipid sink strength that sequesters newly synthesized TAG, thereby relieving the constitutive BADC-dependent inhibition of ACCase reported for WT Arabidopsis. These OLE variants represent novel factors for potentially increasing TAG accumulation in a variety of oil crops.
keywords:
Feedstock Production;Genomics;Lipidomics
published:
2025-11-03
Kim, Min Soo; Choi, Dasol; Ha, Jihyo; Choi, Kyuhyeok; Yu, Jae-Hyuk; Dumesic, James; Huber, George
(2025)
This study shows a new route to produce potassium sorbate (KS) from triacetic acid lactone (TAL), which is a chemical platform that can be biologically synthesized from natural sources. Sorbic acid and its potassium salt (KS) are widely used as preservatives in foods and pharmaceuticals. Three steps are used to produce KS from TAL: 1) hydrogenation of TAL into 4-hydroxy-6-methyltetrahydro-2-pyrone (HMP), 2) dehydration of HMP to parasorbic acid (PSA), and 3) ring-opening and hydrolysis of PSA to KS. TAL can be fully hydrogenated over Ni/SiO2 to give near quantitative yields of HMP. A three-step reaction kinetics model was developed for dehydration of HMP into PSA. This model was used to show that the highest PSA yield occurs at low temperatures. An experimental PSA yield of 84.2% with respect to TAL was obtained which agreed with the prediction of the reaction kinetics model. KOH was used as a coreactant for the ring-opening hydrolysis of PSA to produce >99.9% yield of KS from PSA. Tetrahydrofuran (THF) was used to purify the TAL derived-KS (TAL-KS). The TAL-KS had a KS purity of 95.5%. The overall yield of TAL-KS with respect to TAL was calculated to be 77.3%. TAL-KS produced in this study had similar antimicrobial activities as commercial KS.
keywords:
Conversion;Catalysis;Modeling
published:
2025-11-03
von Haden, Adam C.; Eddy, William; Burnham, Mark B.; Brzostek, Edward; Yang, Wendy; DeLucia, Evan H.
(2025)
Root exudation is a key process for plant nutrient acquisition, but the controls on root exudation and its relationship to soil C and N processes in agroecosystems are unclear. We hypothesized that root exudation rates would be related to root morphological traits, N fertilization, and soil moisture. We also anticipated that root exudation would be correlated with bulk soil enzyme activity. Root exudation, root traits, and bulk soil extracellular enzyme activity were assessed in maize (Zea mays L.), soybean (Glycine max (L.) Merr.), biomass sorghum (Sorghum bicolor (L.) Moench), giant miscanthus (Miscanthus × giganteus), and switchgrass (Panicum virgatum L.). Measurements were taken in situ during two growing seasons with contrasting precipitation regimes, and N fertilization rate was varied in sorghum during one year. Specific root exudation (per unit root surface area) was negatively related to root diameter and was generally higher in annuals than perennials. Sorghum N fertilization did not affect root exudation rates, and soil moisture regime had no effect on annual root exudation rates within maize, sorghum, and miscanthus. Specific root exudation was negatively related to bulk soil C- and N-degrading soil enzyme activities. Intrinsic plant characteristics appeared more important than environmental variables in controlling in situ root exudation rates. The relationships between root diameter, root exudation, and soil C and N processes link root morphological traits to soil functions and demonstrate the potential tradeoffs among plant nutrient acquisition strategies in agroecosystems.
keywords:
Sustainability;Biomass Analytics;Field Data
published:
2025-11-03
Woodruff, William; Deshavath, Narendra Naik; Susanto, Vionna; Rao, Christopher V.; Singh, Vijay
(2025)
Oleaginous yeasts are a promising candidate for the sustainable conversion of lignocellulosic feedstocks into fuels and chemicals, but their growth on these substrates can be inhibited as a result of upstream pretreatment and enzymatic hydrolysis conditions. Previous studies indicate a high citrate buffer concentration during hydrolysis inhibits downstream cell growth and ethanol fermentation in Saccharomyces cerevisiae. In this study, an engineered Rhodosporidium toruloides strain with enhanced lipid accumulation was grown on sorghum hydrolysate with high and low citrate buffer concentrations. Both hydrolysis conditions resulted in similar sugar recovery rates and concentrations. No significant differences in cell growth, sugar utilization rates, or lipid production rates were observed between the two citrate buffer conditions during batch fermentation of R. toruloides. Under fed-batch growth on low-citrate hydrolysate a lipid titer of 16.7 g/L was obtained. Citrate buffer was not found to inhibit growth or lipid production in this engineered R. toruloides strain, nor did reducing the citrate buffer concentration negatively affect sugar yields in the hydrolysate. As this process is scaled-up, $131 per ton of hydrothermally pretreated biomass can be saved by use of the lower citrate buffer concentration during enzymatic hydrolysis.
keywords:
Conversion;Hydrolysate;Lipidomics
published:
2025-11-03
Blanc-Betes, Elena; Gomez-Casanovas, Nuria; Hartman, Melannie D.; Hudiburg, Tara W.; Khanna, Madhu; Parton, William; DeLucia, Evan H.
(2025)
Bioenergy with carbon capture and storage (BECCS) sits at the nexus of the climate and energy security. We evaluated trade-offs between scenarios that support climate stabilization (negative emissions and net climate benefit) or energy security (ethanol production). Our spatially explicit model indicates that the foregone climate benefit from abandoned cropland (opportunity cost) increased carbon emissions per unit of energy produced by 14–36%, making geologic carbon capture and storage necessary to achieve negative emissions from any given energy crop. The toll of opportunity costs on the climate benefit of BECCS from set-aside land was offset through the spatial allocation of crops based on their individual biophysical constraints. Dedicated energy crops consistently outperformed mixed grasslands. We estimate that BECCS allocation to land enrolled in the Conservation Reserve Program (CRP) could capture up to 9 Tg C year–1 from the atmosphere, deliver up to 16 Tg CE year–1 in emissions savings, and meet up to 10% of the US energy statutory targets, but contributions varied substantially as the priority shifted from climate stabilization to energy provision. Our results indicate a significant potential to integrate energy security targets into sustainable pathways to climate stabilization but underpin the trade-offs of divergent policy-driven agendas.
keywords:
Sustainability;Field Data;Modeling
published:
2025-11-03
Blake-Bradshaw, Abigail; Bradshaw, Therin; Beilke, Elizabeth; Gilbert, Andrew; Osborn, Joshua; Fournier, Auriel M.V.
(2025)
Data consist of 55 acoustic recordings collected using Autonomous Recording Units (ARUs) from two locations and sampling periods. Specifically, data include 60-minute WAV files (8 folders, each contains 5 WAV files) from a field trial during February 2025 whereby we shot shotguns at varying distance from ARUs at Emiquon Reserve owned by The Nature Conservancy. Data also include 60-minute WAV files (15 WAV files) from one ARU placed at Big Rice Lake State Fish and Wildlife Area on opening day of waterfowl hunting season during 10-26-2024. Filenames include the ARU ID separated by underscores and the associated date and time e.g., MINI10_20241026_060002.wav was from MINI10 on 10/26/24 at 6 AM.
keywords:
hunting; shotgun; waterfowl; acoustics
published:
2025-10-31
Lopes, Daiane; Dien, Bruce; Hector, Ronald; Singh, Vijay; Thompson, Stephanie R.; Slininger, Patricia J.; Boundy-Mills, Kyria; Jagtap, Sujit; Rao, Christopher V.
(2025)
Rhodotorula toruloides is being developed for the use in industrial biotechnology processes because of its favorable physiology. This includes its ability to produce and store large amounts of lipids in the form of intracellular lipid bodies. Nineteen strains were characterized for mating type, ploidy, robustness for growth, and accumulation of lipids on inhibitory switchgrass hydrolysate (SGH). Mating type was determined using a novel polymerase chain reaction (PCR)-based assay, which was validated using the classical microscopic test. Three of the strains were heterozygous for mating type (A1/A2). Ploidy analysis revealed a complex pattern. Two strains were triploid, eight haploid, and eight either diploid or aneuploid. Two of the A1/A2 strains were compared to their parents for growth on 75%v/v concentrated SGH. The A1/A2 strains were much more robust than the parental strains, which either did not grow or had extended lag times. The entire set was evaluated in 60%v/v SGH batch cultures for growth kinetics and biomass and lipid production. Lipid titers were 2.33–9.40 g/L with a median of 6.12 g/L, excluding the two strains that did not grow. Lipid yields were 0.032–0.131 (g/g) and lipid contents were 13.5–53.7% (g/g). Four strains had significantly higher lipid yields and contents. One of these strains, which had among the highest lipid yield in this study (0.131 ± 0.007 g/g), has not been previously described in the literature.
keywords:
Conversion;Hydrolysate;Lipidomics
published:
2025-10-30
Cao, Dang Viet; Luo, Guangbin; Korynta, Shelby; Liu, Hui; Liang, Yuanxue; Shanklin, John; Altpeter, Fredy
(2025)
Metabolic engineering for hyperaccumulation of lipids in vegetative tissues is a novel strategy for enhancing energy density and biofuel production from biomass crops. Energycane is a prime feedstock for this approach due to its high biomass production and resilience under marginal conditions. DIACYLGLYCEROL ACYLTRANSFERASE (DGAT) catalyzes the last and only committed step in the biosynthesis of triacylglycerol (TAG) and can be a rate-limiting enzyme for the production of TAG. In this study, we explored the effect of intron-mediated enhancement (IME) on the expression of DGAT1 and resulting accumulation of TAG and total fatty acid (TFA) in leaf and stem tissues of energycane. To maximize lipid accumulation these evaluations were carried out by co-expressing the lipogenic transcription factor WRINKLED1 (WRI1) and the TAG protect factor oleosin (OLE1). Including an intron in the codon-optimized TmDGAT1 elevated the accumulation of its transcript in leaves by seven times on average based on 5 transgenic lines for each construct. Plants with WRI1 (W), DGAT1 with intron (Di), and OLE1 (O) expression (WDiO) accumulated TAG up to a 3.85% of leaf dry weight (DW), a 192-fold increase compared to non-modified energycane (WT) and a 3.8-fold increase compared to the highest accumulation under the intron-less gene combination (WDO). This corresponded to TFA accumulation of up to 8.4% of leaf dry weight, a 2.8-fold or 6.1-fold increase compared to WDO or WT, respectively. Co-expression of WDiO resulted in stem accumulations of TAG up to 1.14% of DW or TFA up to 2.08% of DW that exceeded WT by 57-fold or 12-fold and WDO more than twofold, respectively. Constitutive expression of these lipogenic “push pull and protect” factors correlated with biomass reduction. Intron-mediated enhancement (IME) of the expression of DGAT resulted in a step change in lipid accumulation of energycane and confirmed that under our experimental conditions it is rate limiting for lipid accumulation. IME should be applied to other lipogenic factors and metabolic engineering strategies. The findings from this study may be valuable in developing a high biomass feedstock for commercial production of lipids and advanced biofuels.
keywords:
Feedstock Production;Lipidomics;Metabolomics
published:
2025-10-30
Dwivedi, Nidhi; Yamamoto, Senri; Zhao, Yunjun; Hou, Guichuan; Bowling, Forrest; Tobimatsu, Yuki; Liu, Chang-Jun
(2025)
Grass lignocelluloses feature complex compositions and structures. In addition to the presence of conventional lignin units from monolignols, acylated monolignols and flavonoid tricin also incorporate into lignin polymer; moreover, hydroxycinnamates, particularly ferulate, cross-link arabinoxylan chains with each other and/or with lignin polymers. These structural complexities make grass lignocellulosics difficult to optimize for effective agro-industrial applications. In the present study, we assess the applications of two engineered monolignol 4-O-methyltransferases (MOMTs) in modifying rice lignocellulosic properties. Two MOMTs confer regiospecific para-methylation of monolignols but with different catalytic preferences. The expression of MOMTs in rice resulted in differential but drastic suppression of lignin deposition, showing more than 50% decrease in guaiacyl lignin and up to an 90% reduction in syringyl lignin in transgenic lines. Moreover, the levels of arabinoxylan-bound ferulate were reduced by up to 50%, and the levels of tricin in lignin fraction were also substantially reduced. Concomitantly, up to 11 μmol/g of the methanol-extractable 4-O-methylated ferulic acid and 5–7 μmol/g 4-O-methylated sinapic acid were accumulated in MOMT transgenic lines. Both MOMTs in vitro displayed discernible substrate promiscuity towards a range of phenolics in addition to the dominant substrate monolignols, which partially explains their broad effects on grass phenolic biosynthesis. The cell wall structural and compositional changes resulted in up to 30% increase in saccharification yield of the de-starched rice straw biomass after diluted acid-pretreatment. These results demonstrate an effective strategy to tailor complex grass cell walls to generate improved cellulosic feedstocks for the fermentable sugar-based production of biofuel and bio-chemicals.
keywords:
Feedstock Production;Biomass Analytics;Genome Engineering
published:
2025-10-30
Yang, Boming; Yang, Pan; Golub, Emma; Cai, Ximing
(2025)
The lack of farmers’ willingness to grow perennial bioenergy crops (PBCs) presents a critical barrier to the emergence of cellulosic biofuel production. The willingness relies on a complex network of economic, environmental, and social drivers, among which the influence of social factors (e.g., the influence of neighborhood, community, and communication) is less understood. This study addresses this knowledge gap via a survey analysis of midwestern farmers. The survey data are analyzed through ordinary least square regression and structural equation model, which together investigate the individual and interactive impacts of multiple factors on farmers’ decisions to adopt PBCs. Based on a farm-scale analysis, six statistically significant predictors of farmer willingness to grow PBCs are identified: perception of PBCs’ environment benefits, education level, willingness to take risks, familiarity with PBCs, portion of peers already growing PBCs, and support of biorefineries locating in the local community. Among these, the latter three predictors are social support variables. It is found that familiarity with the crops is the most significant predictor of willingness; familiarity is also an important intermediate variable that mediates the influence of many other predictors. In addition, peer adoption can both directly and indirectly affect willingness via its influence on familiarity. These findings suggest that it is a pressing need to improve farmers’ knowledge of PBCs to promote the adoption of such crops.
keywords:
Sustainability;Economics
published:
2025-10-30
Koh, Hyun Gi; Kim, Jinhong; Rao, Christopher V.; Park, Sung-Jin; Jin, Yong-Su
(2025)
A small and efficient DNA mutation-inducing machine was constructed with an array of microplasma jet devices (7 × 1) that can be operated at atmospheric pressure for microbial mutagenesis. Using this machine, we report disruption of a plasmid DNA and generation of mutants of an oleaginous yeast Rhodosporidium toruloides. Specifically, a compact-sized microplasma channel (25 × 20 × 2 mm3) capable of generating an electron density of greater than 1013 cm–3 was constructed to produce reactive species (N2*, N2+, O, OH, and Hα) under helium atmospheric conditions to induce DNA mutagenesis. The length of microplasma channels in the device played a critical role in augmenting both the volume of plasma and the concentration of reactive species. First, we confirmed that microplasma treatment can linearize a plasmid by creating nicks in vitro. Second, we treated R. toruloides cells with a jet device containing 7 microchannels for 5 min; 94.8% of the treated cells were killed, and 0.44% of surviving cells showed different colony colors as compared to their parental colony. Microplasma-based DNA mutation is energy-efficient and can be a safe alternative for inducing mutations compared to conventional methods using toxic mutagens. This compact and scalable device is amenable for industrial strain improvement involving large-scale mutagenesis.
keywords:
Conversion;Genome Engineering
published:
2025-10-29
Zhang, Zhengyi; Feng, Jianqiang; Yang, Chao; Cui, Haiyang (Ocean); Harrison, Wesley; Zhong, Dongping; Wang, Binju; Zhao, Huimin
(2025)
Since the discovery of Hofmann–Löffler–Freytag reaction more than 130 years ago, both the structure and reactivity of nitrogen-centred radicals have been widely studied. Nevertheless, catalytic enantioselective intermolecular radical hydroamination remains a challenge due to the existence of side reactions, the short lifetime of nitrogen-centred radicals and lack of understanding of the fundamental catalytic steps. In the laboratory, nitrogen-centred radicals are produced with radical initiators, photocatalysts or electrocatalysts. In contrast, their generation and reaction are unknown in nature. Here we report a pure biocatalytic system for the photoenzymatic production of nitrogen-centred radicals and enantioselective intermolecular radical hydroaminations by successfully repurposing an ene-reductase through directed evolution. These reactions progress efficiently at room temperature under visible light without any external photocatalysts and exhibit excellent enantioselectivities. A detailed mechanistic study reveals that the enantioselectivity originates from the radical-addition step while the reactivity originates from the ultrafast photoinduced electron transfer from reduced flavin mononucleotide to nitrogen-containing substrates.
keywords:
Conversion;Catalysis
published:
2025-10-29
Chen, Chu-Chun; Dominguez, Francina; Matus, Sean
(2025)
This dataset contains variables from the European Centre for Medium-Range Weather Forecasts (ECMWF) Reanalysis v5 (ERA5; Hersbach et al., 2020). These data were used for the analysis in “The impact of large-scale land surface conditions on the South American low-level jet” published in Geophysical Research Letters.
Acknowledgments:
This work was supported by NSF Award AGS-1852709. We thank Dr. Zhuo Wang and Dr. Divyansh Chug for their valuable feedback and insightful discussions.
References:
Hersbach H, Bell B, Berrisford P, et al. The ERA5 global reanalysis. Q J R Meteorol Soc. 2020; 146: 1999–2049. https://doi.org/10.1002/qj.3803
keywords:
atmospheric sciences; South American low-level jet; land-atmosphere interactions; soil moisture; regional atmospheric circulation; southeastern South America
published:
2025-01-29
Quiroz, Edwin; Ashley, Mary V.; Zaya, David N.
(2025)
These data records weekly aphid and monarch butterfly (Danaus plexippus) neonate counts on individual milkweed plants in multiple raised garden beds in Chicago during the summers of 2023 and 2024. Relationships between aphid infestation and monarch neonates can be investigated along with weekly trends of monarch oviposition and aphid abundances. All gardens included in this study were on the University of Illinois Chicago campus, and within 100 meters of proximity. Data are provided on three milkweed species in 2023, and one milkweed species in 2024.
keywords:
Aphis; Myzocallis; Danaus plexippus; urban gardens; Asclepias syriaca; milkweeds
published:
2025-08-08
Remmers, Justin J.; Allen, Maximilian; Green, Austin M.
(2025)
Count histories from camera traps and remotely sensed covariate data used in N-mixture modeling to assess the site use intensity of raccoons in Illinois.
published:
2025-06-03
Okyem, Samuel; Trinklein, Timothy; Stanislav, Rubakhin; Jonathan, Sweedler
(2025)
This is a peptide imaging data obtained by mtarix assisted laser desoption ionization trapped ion mobility datasets from the central nervous sytem and select ganglion of aplysia Californica.
keywords:
Neuropeptides, Iosmerization, D-amino acids, MALDI-TIMS
published:
2025-07-09
Kim, Ahyoung; Kim, Chansong; Waltmann, Tommy; Vo, Thi; Kim, Eun Mi; Kim, Junseok; Shao, Yu-Tsun; Michelson, Aaron; Crockett, John R.; Kalutantirige, Falon C.; Yang, Eric; Yao, Lehan; Hwang, Chu-Yun; Zhang, Yugang; Liu, Yu-Shen; An, Hyosung; Gao, Zirui; Kim, Jiyeon; Mandal, Sohini; Muller, David; Fichthorn, Kristen; Glotzer, Sharon; Chen, Qian
(2025)
This dataset contains the raw transmission electron microscopy (TEM) and scanning electron microscopy (SEM) images used to calculate the synthesis yield of patchy nanoparticles (NPs), as described in Supplementary Table 1 of the paper “Patchy Nanoparticles by Atomic “Stencilling” (2025).” All the images were taken at the Materials Research Laboratory, University of Illinois at Urbana-Champaign by Qian Chen group.
1. We have 21 subfolders, each with a name corresponding to one of the 21 patchy NPs listed in Supplementary Table 1 of the paper “Patchy Nanoparticles by Atomic “Stencilling” (2025)."
2. In TEM images, the bright and dark regions indicate the polymer patches and NP cores, respectively.
3. In SEM images, the bright and dark regions indicate the NP cores and polymer patches, respectively.
4. Each subfolder contains a “readme (subfolder name).txt” file with more detailed information about each sample.
keywords:
Patchy nanoparticle; polymer; synthesis; self-assembly
published:
2020-06-26
Gasparik, Jessica T.; Ye, Qing; Curtis, Jeffrey H.; Presto, Albert A.; Donahue, Neil M.; Sullivan, Ryan C.; West, Matthew; Riemer, Nicole
(2020)
This dataset contains the PartMC-MOSAIC simulations used in the article "Quantifying Errors in the Aerosol Mixing-State Index Based on Limited Particle Sample Size". The 1000 simulations of output data is organized into a series of archived folders, each containing 100 scenarios. Within each scenario directory are 25 NetCDF files, which are the hourly output of a PartMC-MOSAIC simulation containing all information regarding the environment, particle and gas state. This dataset was used to investigate the impact of sample size on determining aerosol mixing state. This data may be useful as a data set for applying different types of estimators.
keywords:
Atmospheric aerosols; single-particle measurements; sampling uncertainty; NetCDF
published:
2025-10-27
Deshavath, Narendra Naik; Dien, Bruce; Slininger, Patricia J.; Jin, Yong-Su; Singh, Vijay
(2025)
A wide range of inorganic and organic chemicals are used during the pretreatment and enzymatic hydrolysis of lignocellulosic biomass to produce biofuels. Developing an industrially relevant 2G biorefinery process using such chemicals is challenging and requires more unit operations for downstream processing. A sustainable process has been developed to achieve industrially relevant titers of bioethanol with significant ethanol yield. The pretreatment of sorghum biomass was performed by a continuous pilot-scale hydrothermal reactor followed by disk milling. Enzymatic hydrolysis was performed without washing the pretreated biomass. Moreover, citrate buffer strength was reduced to 100-fold (50 mM to 0.5 mM) during the enzymatic hydrolysis. Enzymatic hydrolysis at 0.5 mM citrate buffer strength showed that significant sugar concentrations of 222 ± 2.3 to 241 ± 2.3 g/L (glucose + xylose) were attained at higher solids loadings of 50 to 60% (w/v). Furthermore, hydrolysates were fermented to produce bioethanol using two different xylose-fermenting Saccharomyces cerevisiae strains and a co-culture of xylose-fermenting and non-GMO yeast cultures. Bioethanol titer of 81.7 g/L was achieved with an ethanol yield of 0.48 gp/gs. Additionally, lipids were produced using the oleaginous yeast Rhodosporidium toruloides, yielding 13.2 g/L lipids with cellular lipid accumulation of 38.5% w/w from 100 g/L of sugar concentration. In summary, reducing the strength of the citrate buffer during enzymatic hydrolysis and omitting inorganic chemicals from the pretreatment process enhances the fermentability of hydrolysates and can also reduce operating costs.
keywords:
Conversion;Hydrolysate;Lipidomics
published:
2025-10-27
Cheng, Ming-Hsun; Singh, Shuchi; Carr Clennon, Aidan N.; Dien, Bruce; Singh, Vijay
(2025)
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.
keywords:
Conversion;Biomass Analytics;Feedstock Bioprocessing;Hydrolysate
published:
2025-10-27
Jindra, Michael A.; Choe, Kisurb; Chowdhury, Ratul; Kong, Ryan; Ghaffari, Soodabeh; Sweedler, Jonathan; Pfleger, Brian
(2025)
The dominant strategy for tailoring the chain-length distribution of free fatty acids (FFA) synthesized by heterologous hosts is expression of a selective acyl-acyl carrier protein (ACP) thioesterase. However, few of these enzymes can generate a precise (greater than 90% of a desired chain-length) product distribution when expressed in a microbial or plant host. The presence of alternative chain-lengths can complicate purification in situations where blends of fatty acids are not desired. We report the assessment of several strategies for improving the dodecanoyl-ACP thioesterase from the California bay laurel to exhibit more selective production of medium-chain free fatty acids to near exclusivity. We demonstrated that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) was an effective library screening technique for identification of thioesterase variants with favorable shifts in chain-length specificity. This strategy proved to be a more effective screening technique than several rational approaches discussed herein. With this data, we isolated four thioesterase variants which exhibited a more selective FFA distribution over wildtype when expressed in the fatty acid accumulating E. coli strain, RL08. We then combined mutations from the MALDI isolates to generate BTE-MMD19, a thioesterase variant capable of producing free fatty acids consisting of 90% of C12 products. Of the four mutations which conferred a specificity shift, we noted that three affected the shape of the binding pocket, while one occurred on the positively charged acyl carrier protein landing pad. Finally, we fused the maltose binding protein (MBP) from E. coli to the N – terminus of BTE-MMD19 to improve enzyme solubility and achieve a titer of 1.9 g per L of twelve-carbon fatty acids in a shake flask.
keywords:
Conversion;Genomics
published:
2025-10-24
Choe, Kisurb; Jindra, Michael A.; Hubbard, Susan; Pfleger, Brian; Sweedler, Jonathan
(2025)
Creating controlled lipid unsaturation locations in oleochemicals can be a key to many bioengineered products. However, evaluating the effects of modifications to the acyl-ACP desaturase on lipid unsaturation is not currently amenable to high-throughput assays, limiting the scale of redesign efforts to <200 variants. Here, we report a rapid mass spectrometry (MS) assay for profiling the positions of double bonds on membrane lipids produced by Escherichia coli colonies after treatment with ozone gas. By MS measurement of the ozonolysis products of Δ6 and Δ8 isomers of membrane lipids from colonies expressing recombinant Thunbergia alata desaturase, we screened a randomly mutagenized library of the desaturase gene at 5 s per sample. Two variants with altered regiospecificity were isolated, indicated by an increase in 16:1 Δ8 proportion. We also demonstrated the ability of these desaturase variants to influence the membrane composition and fatty acid distribution of E. coli strains deficient in the native acyl-ACP desaturase gene, fabA. Finally, we used the fabA deficient chassis to concomitantly express a non-native acyl-ACP desaturase and a medium-chain thioesterase from Umbellularia californica, demonstrating production of only saturated free fatty acids.
keywords:
Conversion;Lipidomics;Mass Spectrometry
published:
2025-10-24
Maitra, Shraddha; Singh, Vijay
(2025)
Sweet sorghum is typically cultivated for the food and fodder market. Recently, sweet sorghum varieties are being metabolically transitioned to enhance energy density by accumulating oil droplets in their vegetative tissues for bioenergy applications. Owing to the high biomass yield of sorghum, the transgenic lines can compete with oil-seed crops for biodiesel yield per unit area. In the initial phase of transgenic development, a high-throughput phenotyping method can bridge the gap between the production pipeline and analysis to improve the efficiency of the process. To meet the requirement, the present study extends the application of time-domain 1H-NMR spectroscopy for rapid quantification and characterization of the total in-situ lipids of sweet sorghum ‘ramada’ to lay the groundwork for analyzing the upcoming large quantity of transgenic samples. NMR technology has been successfully established for analyzing lipid contents of vegetative tissues of non-transgenic variety. The multiexponential analysis of spin-lattice (T1) relaxation spectra obtained from TD-NMR aided the investigation of the dynamics of the free and bound lipid fraction with plant development. The total lipid concentration of bagasse and leaves of non-transgenic sweet sorghum remained unchanged throughout the plant development. Leaves displayed a higher percentage of bound lipids as compared to bagasse. A significant variation in the lipid concentration of juice was observed at the different growth stages with a maximum lipid accumulation of 1.21 ± 0.04% w/w at the boot stage that decreased with further maturity of the plant.
keywords:
Conversion;Biomass Analytics;Lipidomics;Metabolomics
planned publication date:
2026-01-01
Iacaruso, Nicholas J.; Myers, Jared T.; Seider, Michael J.; Davis, Mark
(2026)
This dataset contains the data related to Chapter 2 of Iacaruso, N. (2026) "EVALUATING ENVIRONMENTAL DNA AS AN EARLY DETECTION METHOD FOR AQUATIC INVASIVE SPECIES". Doctoral Dissertation. University of Illinois Urbana-Champaign. This chapter will also be represented in Iacaruso et al. (2025) "Environmental DNA metabarcoding for monitoring fish biodiversity in remote lakes". North American Journal of Fisheries Management. (Forthcoming). The files contain the eDNA metabarcoding sequences from sampling Isle Royale lakes in 2021 and 2022, species read counts for each eDNA sample, and other information collected at each site.
keywords:
eDNA; Fish; Management; Cisco