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At least 37 records · Page 2

Filling the cellulosic bio-economy gap by utilizing a wedge approach combined with stakeholder collaboration

The price gap between the market and breakeven prices of cellulosic biomass for farmers represents a significant barrier to the development of a low-carbon cellulosic bioeconomy. Using a bottom-up, agent-based modeling tool that replicates the behaviors and interactions of key stakeholders, this study analyzes the emergence of a cellulosic bioeconomy at the local scale through a wedge approach that examines an integrated portfolio of multiple policy options, including subsidies for small-scale bioproducts and environmental credits. Here, the role of collaboration among multiple stakeholders, such as biomass producers (farmers), bio-refinery industry, government, and society, is assessed for filling the price gap. Using the Sangamon River Basin as a case study site, we evaluate the effectiveness of the wedge approach by comparing simulation results from multiple scenarios, each incorporating different combinations of bioeconomy wedges, with and without stakeholder collaboration. Results underscore that active collaboration among stakeholders acts as a catalyst enlarging the effectiveness of bioeconomy wedges. Including the carbon credits and environmental value in the policy portfolio is found to bridge the price gap through collective contributions from diverse stakeholders, where the cellulosic biofuel and bioproduct industry plays a pivotal role. Although this study is conducted at the local watershed scale, the methodology and findings offer valuable insights for market development in other watersheds and the potential scaling of local markets to regional and national levels.

09 BIOMASS FUELS

Cellulose and the control of growth anisotropy

The overall aim of the award was to understand how the anisotropy of plant cell expansion is controlled by cellulose microfibrils. As experimental material, the project used roots of the popular model plant species, Arabidopsis thaliana. In general, cellulose governs expansion anisotropy by virtue of its strength and organized deposition. Because of their rigidity and alignment, cellulose microfibrils endow the cell wall with mechanical anisotropy; cell walls deform more easily in the direction perpendicular to the microfibrils compared to the parallel direction. Expansion anisotropy thus depends appreciably on the control of cellulose alignment.

59 BASIC BIOLOGICAL SCIENCES

Evaluation of Enzymatic Hydrolysis of CELSS Wheat Residue Cellulose at a Scale Environment to NASA's KSC Breadboard Project

Biomass processing at the Kennedy Space Center CELSS breadboard project has focused on the evaluation of breadboard-scale enzymatic hydrolysis of wheat residue cellulose (25%, w/w). Five replicate runs of cellulase production by Trichoderma reesei (QM9414) and enzymatic hydrolysis of residue cellulose were completed. Enzymes were produced in 1 0 days (5 L, 25 g (dry weight) residue). Cellulose hydrolysis (12 L, 50 g (dry weight) residue) using these enzymes produced 5.5 to 6.0 g glucose liter(exp -1) in 7 days. Cellulose conversion efficiency was 29%. These processes are feasible technically on a breadboard scale, but would only increase the edible wheat yield 10%.

Strayer, Richard F.

Empirical evidence that glucan-interacting amino acid side chains within the transmembrane channel collectively facilitate cellulose synthase function

The fundamental mechanism of cellulose synthesis is widely conserved across Kingdoms and depends on cellulose synthases, which are processive, dual-function, family 2 glycosyltransferases (GT-2). These enzymes polymerize glucose on the cytoplasmic side of the plasma membrane and export the glucan chain to the cell surface through an integral transmembrane (TM) channel. Structural studies of active plant cellulose synthases (CESAs) have revealed interactions between the nascent glucan chain and the side chains of polar, charged, and aromatic amino acid residues that line the TM channel. However, the functional consequences of modifying these side chains have not been tested in vivo in CESAs or other processive GT-2s. To test this, we used an established in vivo assay based on genetic complementation of CESA5 in the moss, Physcomitrium patens. For accurate prediction of glucan-interacting amino acid residues, we generated a complete homotrimeric molecular model of PpCESA5 using a combination of homology and de novo modeling. All-atom molecular dynamics-based analyses of contact metrics and interaction energy identified 23 amino acid residues with high propensity to interact with the nascent glucan chain within the TM channel or on the apoplastic surface of PpCESA5. Mutating any one of 18 of these amino acid residues to alanine, thereby removing their side chains, abolished or impaired CESA function, with the strongest effects observed upon the loss of charged amino acid side chains. This provides direct evidence to support the hypothesis that multiple amino acid residues collectively maintain a smooth energy landscape within the TM channel to facilitate glucan translocation.

59 BASIC BIOLOGICAL SCIENCES

Coarse-grained molecular dynamics simulation of solvent-dependent cellulose nanofiber interactions

Associations between cellulose are important both in biofuel production and in the use of cellulose for biomaterials. Cellulose nanofibers (CNFs) are sustainable, strong, light-weight alternatives to traditional materials in manufacturing, but are challenging to obtain due to irreversible aggregation in solution during preparative fibrillation. Therefore, it is imperative to understand the underlying factors driving aggregation with a view to designing solvents that can effectively compete with interfiber interactions, hence reducing aggregation. Molecular dynamics (MD) simulation at atomic detail can provide useful information on local interactions. However, the length and timescales accessible are too short to fully capture association processes. Here, we provide a method for accessing the longer length and timescales required using coarse-grained (CG) MD simulations with a MARTINI force field to calculate the interaction behavior of CNFs in three selected solvents: NaOH-urea-water, acetone, and neat water. The CG results are consistent with our prior all-atom MD and with previous experimental results. While acetone is found not to be an effective solvent, urea and ionic moieties in NaOH-urea-water not only solvate the fibrils but also improve the confinement of water molecules around them as shown by the solvent residence times and mean-square displacements. Overall, the presence of urea and ions reduces the likelihood of aggregation in multi-CNF systems relative to neat water irrespective of whether the hydrophobic or hydrophilic CNF surfaces are interacting. In conclusion, the CG method shows clear promise for selecting potential high-performance solvents for experimental prioritization in bioenergy and biomaterials research in a relatively fast manner as well as for understanding the aggregation and rheological behavior of CNF-solvent systems.

aggregation

Methodology for Determination of Cellulosic Glucans and Hemicellulose Content in a Fuel Ethanol Production Facility (CRADA Final Report)

Develop a single methodology for determination of the total cellulosic glucan, including both cellulose and beta-glucans, along with hemicellulose content in corn kernel fiber which could be utilized by the dry grind corn ethanol production industry to determine the cellulosic converted fraction of the corn kernel fiber. The work will build upon a methodology already described in literature but will specifically focus on a more complete determination of the structural polysaccharides in corn kernel fiber to make it more applicable to the corn ethanol industry.

09 BIOMASS FUELS

Engineering Clostridium thermocellum for production of 2,3-butanediol from cellulose

Clostridium thermocellum is a promising host for consolidated bioprocessing due to its ability to directly ferment cellulose into fuels and chemicals. However, natural product formation in this organism is limited. Here, we report engineering C. thermocellum for the production of 2,3-butanediol (23BD), a valuable industrial chemical. We functionally expressed a thermophilic 23BD pathway in this organism resulting in a 23BD titer of 19.7 mM from cellulose, representing a metabolic yield of 24%. We used a cell-free systems biology approach to identify limiting steps in the 23BD pathway, revealing that exogenous 23BD dehydrogenase (BDH) activity was essential for production, while native acetolactate synthase (ALS) and acetolactate decarboxylase (ALDC) activities were present but limiting in the parent strain. This approach also revealed redox balance limitations. We demonstrated that this improved understanding of redox balance limitations could be used to increase 23BD titer in vivo, showing that adding acetate could be used to increase 23BD yield. This work establishes a foundation for developing C. thermocellum into a robust platform for 23BD production directly from cellulose and highlights the utility of cell-free systems for guiding metabolic engineering in non-model organisms.

09 BIOMASS FUELS

Development of nonflammable cellulosic foams

The development of a moldable cellulosic foam for use in Skylab instrument storage cushions is considered. Requirements include density of 10 lb cu ft or less, minimal friability with normal handling, and nonflammability in an atmosphere of 70 percent oxygen and 30 percent nitrogen at 6.2 psia. A study of halogenated foam components was made, including more highly chlorinated binders, halogen-containing additives, and halogenation of the cellulose. The immediate objective was to reduce the density of the foam through reduction in inorganic phosphate without sacrificing flame-retarding properties of the foams. The use of frothing techniques was investigated, with particular emphasis on a urea-formaldehyde foam. Halogen-containing flame retardants were deemphasized in favor of inorganic salts and the preparation of phosphate and sulphate esters of cellulose. Utilization of foam products for civilian applications was also considered.

Luttinger, M.

The case for cellulose production on Mars

From examining the consequences of not requiring that all wastes from life support be recycled back to the food plants, it is concluded that cellulose production on Mars could be an important input for many nonmetabolic material requirements on Mars. The fluxes of carbon in cellulose production would probably exceed those in food production, and therefore settlements on Mars could utilize cellulose farms in building a Mars infrastructure.

Volk, Tyler

Microbial Cellulose Assembly in Microgravity

Based on evidence indicating a possible correlation between hypo-gravity conditions and alteration of cellulose production by the gram negative bacterium, Acetobacter xylinum, a ground-based study for a possible long term Space Shuttle flight has been conducted. The proposed experiment for A. xylinum aboard the Shuttle is the BRIC (Biological Research in a Canister), a metal container containing spaces for nine Petri plates. Using a common experimental design, the cellulose production capability as well as the survivability of the A. xylinum strains NQ5 and AY201 have been described. It should now be possible to use the BRIC for the first long term microgravity experiments involving the biosynthesis of cellulose.

Brown, R. Malcolm, Jr.

Light Weight Biomorphous Cellular Ceramics from Cellulose Templates

Bimorphous ceramics are a new class of materials that can be fabricated from the cellulose templates derived from natural biopolymers. These biopolymers are abundantly available in nature and are produced by the photosynthesis process. The wood cellulose derived carbon templates have three- dimensional interconnectivity. A wide variety of non-oxide and oxide based ceramics have been fabricated by template conversion using infiltration and reaction-based processes. The cellular anatomy of the cellulose templates plays a key role in determining the processing parameters (pyrolysis, infiltration conditions, etc.) and resulting ceramic materials. The processing approach, microstructure, and mechanical properties of the biomorphous cellular ceramics (silicon carbide and oxide based) have been discussed.

Singh, Mrityunjay

Processes for treating cellulosic material

Disclosed are processes for pretreating cellulosic materials in liquid water by heating the materials in liquid water at a temperature at or above their glass transition temperature but not substantially exceeding 220.degree. C., while maintaining the pH of the reaction medium in a range that avoids substantial autohydrolysis of the cellulosic materials. Such pretreatments minimize chemical changes to the cellulose while leading to physical changes which substantially increase susceptibility to hydrolysis in the presence of cellulase.

Ladisch, Michael R.

Harnessing plant‐based platform for low‐cost cellulosic sugar recovery from bioenergy crops

The price of pure cellulase enzyme for the recovery of fermentable cellulosic sugars is one of the major challenges that limit the commercialization of second-generation biofuels and bio-based products. This work shows a means to greatly reduce the cost of cellulases. The abundant capacity of plants to synthesize and hyperaccumulate transgenic proteins in their plastids has been demonstrated for two gene constructs using either NPT II or Tet C to regulate the expression of bacterial cellulase Cel6A. Previously, the expression of Cel6A to 20–35% of total soluble leaf protein in tobacco was shown in two consecutive field trials. Moving forward, the key challenge was the extraction of active enzymes from the transgenic tobacco leaves hyperaccumulating bacterial cellulases in a cost-effective manner. The study showed that unpurified crude extracts of NPT II and Tet C transgenic tobacco leaves recovered ~23% and ~29% w/w cellulosic sugars from energycane bagasse, respectively. A supplementation of as low as 25% of purified commercial cellulase improved the glucose recovery by 2.9 times (~85% w/w) compared to 100% crude extract, which is comparable to the glucose recovery obtained by commercial cellulases, thereby suggesting a reduced requirement of commercial cellulases leading to a cost reduction of 75% for biorefineries. Assuming a stable 40% cellulase yield in total soluble protein under field conditions and multiple harvests (one to three) a year, the study estimates that the potential cost for saccharification of 1 t of lignocellulosic biomass can be reduced to 67–200 USD by using crude leaf extracts of transgenic tobacco.

biofuel

Xylose metabolic engineering of Issatchenkia orientalis for 3-hydroxypropionic acid production from cellulosic hydrolysate without nutrient supplementation

Bioconversion of lignocellulosic biomass offers a promising alternative to petroleum-based chemical production. However, inefficient xylose utilization and toxic compounds in cellulosic hydrolysate limit microbial fermentation, as the hydrolysate contains substantial amounts of xylose in addition to glucose. To address these challenges, we engineered Issatchenkia orientalis to produce 3-hydroxypropionic acid (3-HP) directly from sorghum hydrolysate under low-pH conditions. A heterologous xylose utilization pathway consisting of XYL1, XYL2, and XYL3 from Scheffersomyces stipitis was introduced into an engineered 3-HP producing strain, enabling efficient conversion of xylose to 3-HP. The engineered strain produced 46.8 g/L 3-HP from sorghum hydrolysate without nutrient supplementation. To eliminate the lag phase under low-pH conditions, fermentation was conducted at pH 6.0 for the first three days, after which pH control was discontinued and in situ 3-HP accumulation buffered the culture. This partial pH control strategy increased 3-HP productivity by 55% from 0.20 to 0.31 g/L∙h, while maintaining low-pH conditions. Introducing an additional copy of XYL2 further increased 3-HP titer to 53.5 g/L and the yield by 33%, from 0.30 to 0.40 g/g sugars, with pH reaching 4.5 at the end of fermentation. This represents one of the highest reported 3-HP titers and yields from cellulosic hydrolysate without additional nutrient supplementation. This work demonstrates a nutrient-independent and low-pH bioprocess for upgrading lignocellulosic hydrolysate into 3-HP, highlighting the industrial potential of engineered xylose-utilizing I. orientalis for sustainable production of platform chemicals from renewable feedstocks.

3-Hydroxypropionic acid

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

Zwitterion Moieties in Polypeptides Synergistically Enhance the Release of Cellulose and Amorphous Polysaccharides from Plant Cell Walls

This study demonstrated that covalently localized zwitterionic moieties in zwitterionic polypeptides (ZIPs) effectively disrupt hydrogen bonds in cellulosic substrates, including filter paper and plant cell wall materials, without significant cytotoxicity. ZIPs with varying densities of zwitterionic side chains were synthesized via the postpolymerization modification of histidinecontaining oligopeptides. The newly developed ZIPs predominantly comprised repeating units with zwitterionically converted side chains. Such ZIPs can cleave multiple hydrogen bonds by anchoring the zwitterionic structure at specific sites, thereby partially dissociating the polysaccharide chains in the cell wall. They are especially effective in dissolving amorphous cellulose, even at low concentrations in aqueous solutions. Importantly, this effect was achieved with minimal cellular toxicity, harnessing the advantages of ionic liquid-like properties while mitigating their high-toxicity limitations. This biofriendly approach to cell wall denaturation highlights a novel method for controlling hydrogen bond networks in polysaccharides and cell walls. These findings indicate a new approach for reducing biomass recalcitrance and developing next-generation biobased materials and fuels derived from plant cell walls.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH

Hybrid Bio-Based Composites: Enabling Cellulose Nanofiber (CNF) Incorporation into Composites via Macroscale Natural Fiber Carriers

Cellulose nanofibers (CNFs) have significant potential in composites as additives to improve mechanical properties, melt rheology, and more. However, agglomeration of CNFs is a key challenge in composite melt processing as obtaining nano-level dispersion of CNFs often requires cost- and energy-intensive processes (e.g., solvent exchange or freeze drying) due to the strong hornification tendencies of CNF. Herein, we avoid these challenges by using a natural fiber carrier method to integrate CNF into thermoplastic composites. Fibers are co-dried to create a hybrid fiber feedstock for compounding in which natural fibers are decorated with dispersed nanofibers. The hybridized fibers result in up to a 24% increase in tensile strength and up to a 35% increase in Young’s modulus compared to composites only containing natural fibers. The lignocellulosic nanofibers are found to outperform their purely cellulosic counterpart, which is theorized to be due to either an increased propensity for fibrillation of the lignocellulosic fibers or the increased hydrophobicity of the fibers due to the presence of lignin. Surface analysis of fiber feedstocks, via streaming potential measurements and dynamic light scattering (DLS), confirmed a significant change in the feedstock hydrophobicity before and after hybridization. While mild additions of CNF (1 wt.% on the macroscale fiber) do not impact the composite melt viscosity, the viscosity is found to increase at higher CNF loadings (5 wt.% on the macroscale fiber), indicating its utility as a rheology modifier. Lastly, use of these materials as novel feedstocks for medium-scale additive manufacturing in high-fidelity part production was demonstrated.

bio-based

Polyimide Cellulose Nanocrystal Composite Aerogels

Polyimide (PI) aerogels are highly porous solids having low density, high porosity and low thermal conductivity with good mechanical properties. They are ideal for various applications including use in antenna and insulation such as inflatable decelerators used in entry, decent and landing operations. Recently, attention has been focused on stimuli responsive materials such as cellulose nano crystals (CNCs). CNCs are environmentally friendly, bio-renewable, commonly found in plants and the dermis of sea tunicates, and potentially low cost. This study is to examine the effects of CNC on the polyimide aerogels. The CNC used in this project are extracted from mantle of a sea creature called tunicates. A series of polyimide cellulose nanocrystal composite aerogels has been fabricated having 0-13 wt of CNC. Results will be discussed.

Aerogels