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At least 415 records · Page 23
All sourcesNASA NTRSDOE OSTI
DOE OSTI2026

Comparative genomics provides insights into the cold adaptation of endophytic fungi associated with Deschampsia antarctica

Endophytic fungi from Deschampsia antarctica , the southernmost flowering plant, provide insights into the cold adaptation mechanisms of plant-associated fungi in extreme environments. This study presents the genome sequences and comparative analysis of eight fungal isolates from D. antarctica leaves. These Antarctic fungal isolates were analyzed alongside 121 plant-associated fungal genomes to uncover signatures of adaptation and endophytic specialization. Antarctic endophytes show striking patterns, including reduced genome size (∼26.3 Mb on average), streamlined gene content (∼8844 genes), and notably small secretomes (∼288 proteins). Despite this reduced gene repertoire, they maintain a robust set of genes encoding carbohydrate-active enzymes (CAZymes) but lack those for lignin and bacterial cell wall degradation, indicating a symbiotic lifestyle that avoids host damage and predation. One isolate, Alternaria sp. UNIPAMPA017 stood out, with 26% of its genome occupied by transposable elements. Lifestyle, rather than phylogeny, was the main driver of CAZyme and secretome profiles, underscoring ecological convergence. Compared to endophytes from Arabidopsis and Populus, D. antarctica endophytes harbor fewer pectin-degrading enzymes, reflecting their adaptation to the cell wall structure of their monocot host. Together, these fungi reveal a pattern of genomic reduction and functional fine-tuning, hallmarks of life adapted to persist in cold, nutrient-scarce niches.

Ascomycota↗
DOE OSTI2020

Characterization of two types of polysaccharides from Eremurus hissaricus roots growing in Tajikistan

Eremurus Hissaricus (E.hissaricus) is a native plant of Tajikistan that has been used in traditional medicine and whose raw components have been identified as having potential for the domestic economy. We have found that carbohydrates comprise 80–85% of the root biomass, from which we have characterized two types of polysaccharides: A water-soluble polysaccharide (WSP) and an acid-soluble polysaccharide (ASP), with 23% and 8% yields, respectively. The results from FTIR, HPAEC-PAD, HPSEC, methylation linkage analysis and NMR spectroscopy ( 1 H, 13 C, zTOCSY, HSQC, HSQCTOXY and HMBC) demonstrated that WSP has a backbone of (1 → 4)-linked β-d-mannopyranosyl and β-d-glucopyranosyl residues in a 1.5:1 M ratio, having acetylated groups, with weight-average molecular weights of 495 kDa. Analyses of ASP indicate that it is comprised of d-galactose, d-mannose, and d-glucose in a molar ratio of 1.8: 1.6: 1.0, which is consistent with a branched galactoglucomannan polysaccharide with (1 → 4)-linked β-pyranose configurations, and weight-average molecular weight of 179 kDa. The FTIR absorption bands in the region of 875–890 cm 1 and NMR spectroscopy data confirm the presence of β-pyranose configuration of main polysaccharide backbone in the both ASP and WSP samples. These polysaccharides are an available raw material with the potential for use in a number of industrial applications.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Carrot rhamnogalacturonan I structure and composition changed during 2017 in California

Detailed composition, fine structure, and function were determined for carrot pomace fiber remaining after commercial juice extraction from carrots harvested at four different times during 2017, in different California, USA locations. This pomace was treated with commercial enzymes and the changes in carbohydrate composition were analyzed. The enzyme-treated carrot pomace contained pectic oligosaccharides that were branched, unsaturated, partially acetylated and methylated rhamnogalacturonan I (RG-I) with arabinogalacto-oligosaccharide side chains as well as free galacto-oligosaccharides and galacturono-oligosaccharides detected by MALDI-TOF MS and NMR analysis. The molecular weight and degree of galacto-oligosaccharide polymerization decreased while water solubility increased following enzymatic treatment. While carrot soluble dietary fiber remained relatively constant, insoluble dietary fiber and total dietary fiber increased during the year. The carrot pomace fiber structure changed between the June and September harvests so that it was more susceptible to enzymatic degradation, and its shape became more linear. The diversity of carrot RG-I oligosaccharide structures was the greatest in the enzyme-treated pomace harvested in September and the RG-I became almost completely debranched by the enzyme treatment for the pomace harvested in December. The structural properties of these carrot pomace fiber fractions suggest that they can function as a low viscosity beverage food ingredient, and the rhamnogalacturonan I structure indicated the potential for anti-viral, immuno-modulatory and prebiotic properties. Furthermore, the rhamnogalacturonan structural composition changes during the growing season highlights the need for bioactivity monitoring of carrot pomace obtained from different sources, and harvested at different times during the year.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Ferric chloride aided peracetic acid pretreatment for effective utilization of sugarcane bagasse

The synergetic impacts of ferric chloride aided peracetic acid (FPA) pretreatment were investigated to enhance the total biomass utilization through effective cellulose conversion and high-quality lignin production. The sugarcane bagasse pretreatment with 2% peracetic acid (PAA) and 0.1 mol/L ferric chloride (FeCl 3 ) effectively removed 57.3% of lignin and 72.2% of xylan while preserving ~ 97% of cellulose from sugarcane bagasse under mild temperature (90 °C). The FPA pretreated sugarcane bagasse was effectively hydrolyzed with a glucose yield of 313.0 mg/g-biomass, which was 4.5 times higher than the yield of untreated biomass (69.75 mg/g-biomass) and 1.6 and 3.6 times higher than that of individual PAA and FeCl 3 pretreated sugarcane bagasse, respectively. The regenerated lignin (FPA lignin) showed great potential for further valorization by preserving the major interunit linkage (up to 86% of β-O-4) without significant carbohydrate contamination and lignin condensation due to its mild reaction conditions. In this paper, the combination of PAA and FeCl 3 synergistically enhanced the pretreatment efficiency on sugarcane bagasse and resulted in high fermentable sugar and high-quality lignin production.

09 BIOMASS FUELS↗
DOE OSTI2022

Combustion dynamics of crude and upgraded Thermal DeOxygenation oils in a compression ignition engine

Thermal DeOxygenation (TDO) is a robust thermochemical conversion scheme to produce hydrocarbons from biomass feedstock. The process targets the carbohydrate fraction of biomass to yield a broad mixture of primarily aromatic hydrocarbons within a boiling point range of 348–798 K with low oxygen content (<4 wt%). The resulting materials are amenable to traditional hydrotreating and distillation processes with approximately 70% by mass in the distillate fuel range. The simple conversion scheme combined with commercially viable upgrading routes makes TDO oils a feasible alternative fuel for transportation applications. Here this paper is the first systematic treatment of fit-for-purpose testing of TDO oils in a compression ignition engine. Blends of partially upgraded TDO oils (e.g. whole oil, distilled oil, hydrotreated oil and hydrotreated-distilled oil) are prepared with certified ultra-low sulfur diesel at 5%, 10%, 15% and 20% by volume. The resulting fuels are analyzed for fuel characteristics and combustion dynamics in an instrumented single-cylinder compression ignition engine. All fuel blends at 10% blend level by volume exhibited adequate engine performance. Hydrotreating of the oils is a necessary step to meet cetane number and EPA soot emissions requirements at 20% blend volume. Combined distillation and hydrotreatment meet or exceed all fuel specification and engine performance benchmarks. Partially upgraded TDO oils are suitable fuel options for compression ignition applications.

42 ENGINEERING↗
DOE OSTI2025

Impact of non-irrigation on 1G and 2G bioethanol potential of oilcane feedstock: A field to fuel pipeline study

This study evaluates the bioethanol potential in response to irrigation (IR) and non-irrigation (NIR) of oilcane (OC) during a seasonal drought prior harvest. The juice was extracted through mechanical pressing of stems and fermented by Ethanol Red® yeast to produce first-generation bioethanol. Hydrothermal pretreatment followed by enzymatic hydrolysis of bagasse was performed to produce monomeric sugars from structural carbohydrates. The hydrolysates were fermented with engineered yeast for second-generation bioethanol production. The irrigated oilcane juice (276.3 ± 8.9 g/L) constitutes higher sugar concentrations than non-irrigated oilcane juice (236.5 ± 2.2 g/L). The enzymatic hydrolysis of IR-OC and NIR-OC pretreated bagasse yielded similar concentrations of 247.5 ± 2.22 and 249.7 ± 4.98 g/L fermentable sugars. Industry-relevant bioethanol titers of ≥99 g/L and ≥75 g/L were achieved from juice and hydrolysates, respectively. Therefore, the non-irrigation regime did not impact the 1G and 2G bioethanol titers. However, the overall bioethanol yield can be lower due to the reduction of stem yield (8 %) per hectare.

09 BIOMASS FUELS↗
DOE OSTI2024

Sustainable aviation fuels from biomass and biowaste via bio- and chemo-catalytic conversion: Catalysis, process challenges, and opportunities

Sustainable aviation fuel (SAF) production from biomass and biowaste streams is an attractive option for decarbonizing the aviation sector, one of the most-difficult-to-electrify transportation sectors. Despite ongoing commercialization efforts using ASTM-certified pathways (e.g., lipid conversion, Fischer-Tropsch synthesis), production capacities are still inadequate due to limited feedstock supply and high production costs. New conversion technologies that utilize lignocellulosic feedstocks are needed to meet these challenges and satisfy the rapidly growing market. Combining bio- and chemo-catalytic approaches can leverage advantages from both methods, i.e., high product selectivity via biological conversion, and the capability to build C-C chains more efficiently via chemical catalysis. Herein, conversion routes, catalysis, and processes for such pathways are discussed, while key challenges and meaningful R&D opportunities are identified to guide future research activities in the space. Bio and chemo-catalytic conversion primarily utilize the carbohydrate fraction of lignocellulose, leaving lignin as a waste product. This makes lignin conversion to SAF critical in order to utilize whole biomass, thereby lowering overall production costs while maximizing carbon efficiencies. Thus, lignin valorization strategies are also reviewed herein with vital research areas identified, such as facile lignin depolymerization approaches, highly integrated conversion systems, novel process configurations, and catalysts for the selective cleavage of aryl C–O bonds. The potential efficiency improvements available via integrated conversion steps, such as combined biological and chemo-catalytic routes, along with the use of different parallel pathways, are identified as key to producing all components of a cost-effective, 100% SAF.

09 BIOMASS FUELS↗
DOE OSTI2023

Graphene oxide exposure alters gut microbial community composition and metabolism in an in vitro human model

Graphene oxide (GO) nanomaterials have unique physicochemical properties that make them highly promising for biomedical, environmental, and agricultural applications. Despite the increasing interest and the use of GO, assessments of its nanotoxicity have largely not interrogated its potential impact on the gut microbiome. This study addresses an important knowledge gap by investigating the impact of GO exposure- both at low (25 ppm) and high (250ppm) doses and fed (nutrient rich) and fasted (nutrient deplete) conditions- on the gut microbial community structure and function, using an in vitro human colon bioreactor model. 16S rRNA amplicon sequencing revealed that GO exposure resulted in a restructuring of community composition. 25 ppm GO induced a marked decrease in the Bacteroidota phylum and increased the ratio of Firmicutes to Bacteroidota (F/B). Untargeted metabolomics on the supernatants indicated that 25 ppm GO impaired microbial utilization and metabolism of substrates (amino acids, carbohydrate metabolites) and reduced production of beneficial microbial metabolites such as 5-hydroxyindole-3-acetic acid and GABA. Exposure to 250 ppm GO resulted in community composition and metabolome profiles that were very similar to the controls that lacked both GO and digestive enzymes, suggesting that high concentrations of GO may interact with digestive enzymes to form protein coronas, causing their depletion in the gut environment. Differential abundance analyses revealed that 3 genera from the phylum Bacteroidota (Bacteroides, Dysgonomonas, and Parabacteroides) were more abundant after 250 ppm GO exposure, irrespective of feed state. Integrative correlation network analysis indicated that the phylum Bacteroidota showed strong positive correlations to multiple microbial metabolites including GABA and 3-indoleacetic acid, are much larger number of correlations compared to other phyla. These results show that GO exposure has a significant impact on gut microbial community composition and metabolism and different mechanisms are at play for low and high GO concentrations.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Effects of the advanced organosolv pretreatment strategies on structural properties of woody biomass

Plants have evolved their lignocellulosic cell walls through complex structural and chemical mechanisms to protect itself against microbial attacks, which makes native lignocellulosic biomass recalcitrant to enzymatic deconstruction. Pretreatment is a crucial step in the biological conversion of biomass as it can render structural changes in the plant cell wall to reduce the biomass recalcitrance, thus enhancing its sugar release performance. There have been many efforts to develop effective pretreatment technologies to overcome the biomass recalcitrance with a primary focus on the efficient conversion of biomass carbohydrates to liquid fuels, while lignin is significantly underutilized despite its bulk amount and high-value opportunities. In this study, the effects of two recent organosolv pretreatment strategies, co-solvent enhanced lignocellulosic fractionation (CELF) and γ-Valerolactone (GVL) pretreatments, on physicochemical properties of poplar were investigated and compared with the effects of conventional ethanol organosolv pretreatment. Diverse physicochemical properties of biomass including chemical compositions, molecular weights of cellulose and lignin, aromatics and inter-unit linkages of lignin, lignin hydroxyl group contents, cellulose crystallinity, and accessible surface area of cellulose were analyzed before and after pretreatments. The results revealed how each organic solvent pretreatment system affected biomass structural characteristics and recalcitrance.

Meng, Xianzhi↗
DOE OSTI2024

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 1 H 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.

09 BIOMASS FUELS↗
DOE OSTI2026

Synergistic effects of chemical-free hydrothermal pretreatment on the recovery of cellulosic sugars and pectin from sugar mill-derived sugar beet pulp

Sugar mills processing sugar beet generate large amounts of sugar beet pulp (SBP), a carbohydrate-rich byproduct composed of 22–30% cellulose, 24–32 % hemicellulose, and 15-20% pectin. Mild, chemical-free hydrothermal pretreatment offers a promising approach to recover these compounds while minimizing pectin degradation, reducing chemical use, and lowering operating costs for value-added applications. This study evaluates the effects of chemical-free hydrothermal pretreatment on cellulosic sugars recovery and its synergy with subsequent pectin extraction under pretreatment conditions ranging from 80°C to 120°C for 15-45 min. Results show that increasing pretreatment severity preserved most glucans with 4% decrease in galacturonan content. Optimal pretreatment (100°C for 45 min) followed by enzymatic hydrolysis achieved the best glucose (95%) and pentose (74%) yields, while galacturonan remained concentrated in the residual solids. Subsequent citric acid extraction at 80°C for 3 hr and a solid-to-liquid ratio of 1:15 yielded up to 85% pectin, significantly lowering water, chemical, and energy requirements compared to conventional industrial extraction. The integration of mild hydrothermal pretreatment with enzymatic hydrolysis thus maximized sugar recovery and enabled efficient downstream pectin extraction without compromising product yields. These findings advance low-impact valorization strategies to reinforce SBP’s values in biorefinery.

09 - BIOMASS FUELS↗
DOE OSTI2023

Lytic polysaccharide monooxygenase synergized with lignin-degrading enzymes for efficient lignin degradation

Even though the discovery of lytic polysaccharide monooxygenases (LPMOs) has fundamentally shifted our understanding of biomass degradation, most of the current studies focused on their roles in carbohydrate oxidation. However, no study demonstrated if LPMO could directly participate to the process of lignin degradation in lignin-degrading microbes. This study showed that LPMO could synergize with lignin-degrading enzymes for efficient lignin degradation in white-rot fungi. The transcriptomics analysis of fungi Irpex lacteus and Dichomitus squalens during their lignocellulosic biomass degradation processes surprisingly highlighted that LPMOs co-regulated with lignin-degrading enzymes, indicating their more versatile roles in the redox network. Biochemical analysis further confirmed that the purified LPMO from I. lacteus CD2 could use diverse electron donors to produce H 2 O 2 , drive Fenton reaction, and synergize with manganese peroxidase for lignin oxidation. The results thus indicated that LPMO might uniquely leverage the redox network toward dynamic and efficient degradation of different cell wall components.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Insights into the dual cleavage activity of the GH16 laminarinase enzyme class on β-1,3 and β-1,4 glycosidic bonds

Glycoside hydrolases (GH) are involved in the degradation of a wide diversity of carbohydrates and present several biotechnological applications. Many GH families are composed of enzymes with a single well-defined specificity. In contrast, enzymes from the GH16 family can act on a range of different polysaccharides, including β-glucans and galactans. SCLam, a GH16 member derived from a soil metagenome, an endo-β-1,3(4)-glucanase (EC 3.2.1.6), can cleave both β-1,3 and β-1,4 glycosidic bonds in glucans, such as laminarin, barley β-glucan, and cello-oligosaccharides. A similar cleavage pattern was previously reported for other GH16 family members. However, the molecular mechanisms for this dual cleavage activity on (1,3)- and (1,4)-β-D-glycosidic bonds by laminarinases have not been elucidated. In this sense, we determined the X-ray structure of a presumably inactive form of SCLam co-crystallized with different oligosaccharides. The solved structures revealed general bound products that are formed due to residual activities of hydrolysis and transglycosylation. Biochemical and biophysical analyses and molecular dynamics simulations help to rationalize differences in activity towards different substrates. Our results depicted a bulky aromatic residue near the catalytic site critical to select the preferable configuration of glycosidic bonds in the binding cleft. Altogether, these data contribute to understanding the structural basis of recognition and hydrolysis of β-1,3 and β-1,4 glycosidic linkages of the laminarinase enzyme class, which is valuable for future studies on the GH16 family members and applications related to biomass conversion into feedstocks and bioproducts.

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

Machine learning reveals sequence-function relationships in family 7 glycoside hydrolases

Family 7 glycoside hydrolases (GH7) are among the principal enzymes for cellulose degradation in nature and industrially. These important enzymes are often bimodular, including a catalytic domain and carbohydrate binding module (CBM) attached via a flexible linker, and exhibit an active site that binds cello-oligomers of up to ten glucosyl moieties. GH7 cellulases consist of two major subtypes: cellobiohydrolases (CBH) and endoglucanases (EG). Despite the critical importance of GH7 enzymes, there remain gaps in our understanding of how GH7 sequence and structure relate to function. Here, we employed machine learning to gain data-driven insights into relationships between sequence, structure, and function across the GH7 family. Machine-learning models, trained only on the number of residues in the active-site loops as features, were able discriminate GH7 CBHs and EGs with up to 99% accuracy, demonstrating that the lengths of loops A4, B2, B3, and B4 strongly correlate with functional subtype across the GH7 family. Classification rules were derived such that specific residues at 42 different sequence positions each predicted the functional subtype with accuracies surpassing 87%. A random forest model trained on residues at 19 positions in the catalytic domain predicted the presence of a CBM with 89.5% accuracy. Our ML results recapitulate, as top-performing features, a substantial number of the sequence positions determined by previous experimental studies to play vital roles in GH7 activity. We surmise that the yet-to-be-explored sequence positions among the top-performing features also contribute to GH7 functional variation and may be exploited to understand and manipulate function.

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

Expression, purification, and characterization of diacylated Lipo-YcjN from Escherichia coli

YcjN is a putative substrate binding protein expressed from a cluster of genes involved in carbohydrate import and metabolism in Escherichia coli. Here, we determine the crystal structure of YcjN to a resolution of 1.95 Å, revealing that its three-dimensional structure is similar to substrate binding proteins in subcluster D-I, which includes the well-characterized maltose binding protein. Furthermore, we found that recombinant overexpression of YcjN results in the formation of a lipidated form of YcjN that is posttranslationally diacylated at cysteine 21. Comparisons of size-exclusion chromatography profiles and dynamic light scattering measurements of lipidated and nonlipidated YcjN proteins suggest that lipidated YcjN aggregates in solution via its lipid moiety. Additionally, bioinformatic analysis indicates that YcjN-like proteins may exist in both Bacteria and Archaea, potentially in both lipidated and nonlipidated forms. Together, our results provide a better understanding of the aggregation properties of recombinantly expressed bacterial lipoproteins in solution and establish a foundation for future studies that aim to elucidate the role of these proteins in bacterial physiology.

Escherichia coli↗
DOE OSTI2021

Economic perspective of ethanol and biodiesel coproduction from industrial hemp

Herein, the economics of producing biofuels from an industrial hemp (Cannabis sativa) genotype e 19m96136 was investigated. A lignocellulosic biofuel plant, hourly consuming 85 metric tons of hemp biomass was modeled in SuperPro Designer®. The integrated bioenergy plant produced hemp biodiesel and bioethanol from lipids and carbohydrates, respectively. The structural composition of the industrial hemp plant was analyzed in a previous study. The data obtained was used to simulate feedstock composition in SuperPro Designer®. The simulation results indicated that Hemp containing 2% lipids can yield up to 3.95 million gallons of biodiesel annually. On improving biomass lipid content to 5 and 10%, biodiesel production increased to 9.88 and 19.91 million gallons, respectively. The breakeven unit production cost of hemp biodiesel with 2, 5, and 10% lipid containing hemp was $18.49, $7.87, and $4.13/gallon, respectively. The biodiesel unit production cost when utilizing 10% lipid-containing hemp was comparable to soybean biodiesel at $4.13/ gallon. Furthermore, sensitivity analysis revealed the possibility of a 7.80% reduction in unit production cost upon a 10% reduction in hemp feedstock cost. Furthermore, industrial hemp was capable of pro- ducing between 307.80 and 325.82 gallons of total biofuels per hectare of agricultural land than soybean.

09 BIOMASS FUELS↗
DOE OSTI2025

Upconversion of non-recycled MSW paper fractions into biochar via slow pyrolysis and life cycle analysis: Pathways to net negative GHG emission

This study presents an integrated and sustainable approach to valorizing non-recycled municipal solid waste (MSW), a heterogeneous and underutilized waste stream destined for landfilling, by converting it into valuable biochar resources. Specifically, we investigated the upcycling of nonrecycled paper waste based on compositional analysis into four major fractions: high cellulose, high lignin, high contamination, and high ash content papers. These fractions were then homogenized and subjected to slow pyrolysis. The high cellulose fraction (36.1 %) was the most abundant, and contained 66.7 % cellulose, while the high lignin fraction showed the highest lignin (12.1 %) and carbon content (44 %), resulting in highest energy value of 17.4 MJ kg −1 . Biochar yields ranged from 25.6 % to 35.6 %, with the high ash fraction producing the highest yield and alkalinity (pH ≈ 11.2) due to its higher mineral content. Elemental analysis revealed enhanced carbon content up to 76.9 % and reduced oxygen and hydrogen, confirming effective carbonization. The high lignin-derived biochar showed the highest aromatic carbon content (82.8 %) and greater structural stability, while contaminated and ash-rich fractions exhibited dense, low-porosity surfaces due to the presence of contaminants and minerals. Spectroscopic analysis revealed degradation of carbohydrates, disappearance of cellulose peaks and formation of aromatic and mineral derived phases. The scaled life cycle process yielded a global warming potential (GWP) of 119.3 kg CO 2 -eq per ton of dry paper waste, offset by soil carbon sequestration of − 556.41 kg CO 2 -eq, resulting in a net impact of − 427.36 kg CO 2 -eq. This represents a net carbon removal exceeding by ~186 % the emissions associated with landfilling paper waste with electricity generation.

09 BIOMASS FUELS↗
DOE OSTI2021

In vivo evolution of lactic acid hyper-tolerant Clostridium thermocellum

Lactic acid (LA) has several applications in the food, cosmetics and pharmaceutical industries, as well as in the production of biodegradable plastic polymers, namely polylactides. Industrial production of LA is essentially based on microbial fermentation. Recent reports have shown the potential of the cellulolytic bacterium Clostridium thermocellum for direct LA production from inexpensive lignocellulosic biomass. However, C. thermocellum is highly sensitive to acids and does not grow at pH < 6.0. Improvement of LA tolerance of this microorganism is pivotal for its application in cost-efficient production of LA. In the present study, the LA tolerance of C. thermocellum strains LL345 (wild-type fermentation profile) and LL1111 (high LA yield) was increased by adaptive laboratory evolution. At large inoculum size (10 %), the maximum tolerated LA concentration of strain LL1111 was more than doubled, from 15 g/L to 35 g/L, while subcultures evolved from LL345 showed 50–85 % faster growth in medium containing 45 g/L LA. Gene mutations (pyruvate phosphate dikinase, histidine protein kinase/phosphorylase) possibly affecting carbohydrate and/or phosphate metabolism have been detected in most LA-adapted populations. Although improvement of LA tolerance may sometimes also enable higher LA production in microorganisms, C. thermocellum LA-adapted cultures showed a yield of LA, and generally of other organic acids, similar to or lower than parental strains. Based on its improved LA tolerance and LA titer similar to its parent strain (LL1111), mixed adapted culture LL1630 showed the highest performing phenotype and could serve as a framework for improving LA production by further metabolic engineering.

59 BASIC BIOLOGICAL SCIENCES↗