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Carbohydrates produced in source tissues (e.g. mature leaves) must be distributed to sink tissues (e.g. roots and reproductive organs) to fuel growth and development and prevent feedback inhibition of photosynthesis in source leaves.
Biomass deconstruction remains integral for enabling second-generation biofuel production at scale. However, several steps necessary to achieve significant solubilization of biomass, notably harsh pretreatment conditions, impose economic barriers to commercialization. By employing hyperthermostable cellulase machinery, biomass deconstruction can be made more efficient, leading to milder pretreatment conditions and ultimately lower production costs. The hyperthermophilic bacterium Caldicellulosiruptor bescii produces extremely active hyperthermostable cellulases, including the hyperactive multifunctional cellulase CbCel9A/Cel48A. Recombinant CbCel9A/Cel48A components have been previously produced in Escherichia coli and integrated into synthetic hyperthermophilic designer cellulosome complexes. Since then, glycosylation has been shown to be vital for the high activity and stability of CbCel9A/Cel48A. Here, we studied the impact of glycosylation on a hyperthermostable designer cellulosome system in which two of the cellulosomal components, the scaffoldin and the GH9 domain of CbCel9A/Cel48A, were glycosylated as a consequence of employing Ca. bescii as an expression host. Inclusion of the glycosylated components yielded an active cellulosome system that exhibited long-term stability at 75 degrees C. The resulting glycosylated designer cellulosomes showed significantly greater synergistic activity compared to the enzymatic components alone, as well as higher thermostability than the analogous nonglycosylated designer cellulosomes. These results indicate that glycosylation can be used as an essential engineering tool to improve the properties of designer cellulosomes. Additionally, Ca. bescii was shown to be an attractive candidate for production of glycosylated designer cellulosome components, which may further promote the viability of this bacterium both as a cellulase expression host and as a potential consolidated bioprocessing platform organism.
Land use change (LUC) CO 2 emissions associated with bioenergy production depend on the amount of land required to produce bioenergy crops, the carbon stored in such crops (including in the leaves, stalk, roots and soil), and the carbon emitted when another land cover is directly or indirectly displaced as a result. In this study, we use a global integrated assessment model [the Global Change Analysis Model (GCAM)] to explore the differences in estimates of LUC CO 2 emissions for two crops (corn and switchgrass) used to produce ethanol in the United States under alternative assumptions about natural lands protection. Varying the latter assumptions for corn ethanol results in net LUC CO 2 emissions between 7 and 41 gCO 2 per MJ of ethanol, whereas varying the same assumptions for switchgrass ethanol results in net emissions between -26 and 14 gCO 2 per MJ of ethanol. The low-end estimate for each occurs when natural lands are assumed to be fully protected everywhere, which leads to significant cropland intensification. The high-end estimate for each occurs when natural lands are assumed to be unprotected everywhere, leading to greater cropland expansion and associated conversion of unmanaged forest and pasture. Results from this study could be used to inform scenarios of future energy system change or life cycle assessment of biofuels for which LUC emissions would be an input.
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The aim of this study was to determine carbohydrate recovery from hemp for ethanol production and quantify biodiesel from TAG (triacylglycerol) present in hemp. The structural composition of five different hemp varieties (Seward County-SC, York County-YC, Loup County-LC, 19 m96136-19 m, and CBD Hemp-CBD) were analyzed. Concentration of glucan and xylan ranged between 32.63 to 44.52% and 10.62 to 15.48% respectively. The biomass was then pretreated with Liquid hot water followed by disk milling and then hydrolyzed enzymatically to yield monomeric sugars. High glucose (63-85%) and xylose (73-88%) recovery was achieved. Lipids were extracted from hemp using hexane and isopropanol and then transesterified to produce biodiesel. Approximately, 50% of total fatty acids in SC, LC, and CBD hemp were linoleic acid. Palmitic acid was present between 32 to 50% in varieties YC and 19 m. Highest TAG concentration at 25% of total lipids was observed in CBD hemp. The analysis on lipid composition and high sugar recovery demonstrates hemp as a potential bioenergy crop for ethanol and biodiesel coproduction.
A novel process applying high solids loading in chemical-free pretreatment and enzymatic hydrolysis was developed to produce sugars from bioenergy sorghum. Hydrothermal pretreatment with 50% solids loading was performed in a pilot scale continuous reactor followed by disc refining. Sugars were extracted from the enzymatic hydrolysis at 10% to 50% solids content using fed-batch operations. Three surfactants (Tween 80, PEG 4000, and PEG 6000) were evaluated to increase sugar yields. Hydrolysis using 2% PEG 4000 had the highest sugar yields. Glucose concentrations of 105, 130, and 147 g/L were obtained from the reaction at 30%, 40%, and 50% solids content, respectively. The maximum sugar concentration of the hydrolysate, including glucose and xylose, obtained was 232 g/L. Additionally, the glucose recovery (73.14%) was increased compared to that of the batch reaction (52.74%) by using two-stage enzymatic hydrolysis combined with fed-batch operation at 50% w/v solids content.
The objective of Liatris’s 3-year DOE BENEFIT project was to develop and validate a high-performance, lightweight, aerogel-based insulation board of ≥R-10/inch at a cost of ≤$\$$1.50 per board foot using non-flammable inorganic nanocomposites. We have summarized each yearly goal below: Year 1 – Ambient dried inorganic/organic nanocomposite, R-6/inch Year 2 – Processing efficiency and cost down, R-8/inch at $\$$2.0/board-ft Year 3 – Scale up to continuous, pilot production of R-10 at ≤ $\$$1.50/board-ft Over the course of the project period, Liatris has successfully met or exceeded all of the goals outlined above. The project scope over 3 years was extensive, covering method development of ambient dried silica aerogels in Budget Period 1, process formulation and R-value improvement in Budget Period 2, and composite scale up with corporate partners in Budget Period 3. Over the course of the project Liatris has been able to progressively improve the thermal resistance properties from about R-6/in to > R-11/in, 2-3X the performance of conventional insulation products.
GTI Energy has integrated the Cool Reformer technology with the IH 2® -50 pilot plant (which can process a maximum of 50kg/day of biomass feed) to demonstrate the suitability of this new reforming technology for producing hydrogen within IH 2 ® process. As part of this study, we showed that all of the hydrogen required for the IH 2® process could be produced from steam reforming the IH 2® light gas generated within the process and the system can be integrated in a cost-effective way compared to the use of standard SMRs.
The purpose of the research project was to reveal the perceptions of residents and communities of the City and County of Honolulu concerning bioenergy projects, feedstock, and sustainable aviation fuel. The perception study was conducted via community-scale surveys, interviews, and town meetings to gather feedback on the preliminary design of a proposed bioenergy project, including feedstock choice and the need for sustainable aviation fuel in Hawaii. Residents on the west side of Oahu were targeted due to their proximity to the proposed plant site location. Findings from this study aim to identify resident perceptions, understanding, and desire for bioenergy projects and new infrastructure to, improve the energy efficiency and sustainability of Hawaiian islands. While Hawaii has created policies and mandates to move toward 100% renewable energy sources by 2045, the focus has been predominately on renewable electricity, largely overlooking other sustainable energy options such as sustainable aviation fuel. There is a lack of research on the community engagement and perceptions of Hawaiian residents on bioenergy projects and sustainable aviation fuel, contributing to the low adoption of bioenergy projects on the islands. Insights from this study aim to add literature on the need for community engagement in the design process, the importance of accepting new sustainable infrastructure, and the production and use of sustainable aviation fuels.
As awareness of climate change increases, the need for carbon neutral fuel sources is growing. Lignocellulosic biofuel derived from pine trees has been suggested as one potential energy source; however, it requires more research before its efficacy for climate change mitigation can be determined. Due to the large share of forest carbon held in soils and the extensive area of pine plantations in the southeast U.S., a better understanding of plantation soil carbon dynamics is critical for biofuel carbon accounting. This study evaluated the effects of canopy development and productivity on soil CO2 efflux, a proxy for soil respiration (Rs), in an intensively managed loblolly pine (Pinus taeda) stand over a period from May 2015 to December 2019. We found that leaf area index (LAI) and gross ecosystem production (GEP), as well as meteorological variables, had significant effects on Rs, but that both overall Rs and soil carbon pools did not increase over the course of the study. We thus hypothesize that GEP and LAI had intra-annual effects on Rs, and that the lack of change in Rs is the result of an increase in autotrophic respiration (Ra) that offset a decrease in decomposition of the previous stand’s organic matter.
Phase II work is meant to develop an advanced understanding of how production of a renewable diesel blendstock from lignocellulosic biomass feedstocks via aerobic respiration at different production scales affects emissions of criteria pollutants and greenhouse gases (GHGs), local and regional air quality. The results could inform decisions by stakeholders (e.g., industry, community, air quality planners, investors) regarding relationships and tradeoffs between multiple environmental impacts and the challenges and opportunities for the design and deployment of a sustainable biofuel supply chain.
Consistent and reliable preprocessing, conveyance, and reactor in-feed systems, particularly for low-cost waste feedstocks, remains a major technical challenge for the emerging Bioeconomy. By identifying critical biomass attributes and connecting them to flow and conversion behavior, science-driven system designs can address these often-overlooked solids handling challenges. The Wonderful Company (TWC) is the world's largest almond and pistachio grower, generating 250,000 dry tons/year of waste material including hulls, shells, and wood (>5 million tons/year industry wide in the U.S.). This project seeks to turn this environmental and economic liability into a sustainable and profitable resource, targeting conversion via gasification to syngas for electricity and bio-char, by addressing related material handling and feeding challenges. Based on optimized preprocessing strategies, bulk material flow, and thermal conversion properties, an overall system design will be developed. The methodology will be tested with FCIC's benchmark loblolly pine residues, to demonstrate the robustness of the overall approach and provide insight and guidance for future systems. The project will culminate in extended field trials to demonstrate an improved continuous feeding system with a commercial biomass-to-electricity gasifier vendor, and an economic analysis demonstrating a reduction in electricity production costs by maximizing on-stream time while minimizing preprocessing and CapEx costs.
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Preparation of deuterium incorporated cellulose is a vital tool to investigate cellulose internal structure and to expand the application fields of cellulose materials. In this study, cellulosic cotton fibers with anti-rehydration (exchange-resistant) deuterium incorporated in cellulose were prepared by chemical hydrogen–deuterium exchange treatment. The chemical hydrogen–deuterium exchange process, along with exchange time, were characterized by nuclear magnetic resonance hydrogen spectroscopy (1H-NMR). The anti-rehydration deuterium incorporation was determined by Fourier Transform infrared spectroscopy (FTIR) and Stable Isotope Ratio Mass Spectrometer (IRSM). Here, the effect of the deuterium hydroxyl substitution on cotton fiber’s spectral data, microstructure, crystalline information, degree of polymerization, as well as it’s thermogravimetric analysis (charcoalization and combustion) are explored. Analysis of the chemical exchange process indicated that the hydrogen–deuterium exchange occurred preferentially in the amorphous cellulose component over the first several minutes. Deuterium exchange in the anti-rehydration crystalline phase took several hours. Increasing the treatment time, enhanced exchange-resistant deuterium incorporation to as high as about 60% of the cotton fibers’ cellulose hydroxyl groups was achieved. The characterization of FTIR, Fourier transform Raman (FT-Raman), and near-infrared spectra (NIR) all exhibited the deuterium spectral isotope effect on cellulose hydroxl groups. While, apart from the effect of reaction temperature, deuterium incorporation isotope effect did not affect the cellulose microstructure, crystalline index and the degree of polymerization properties. Furthermore, the thermogravimetric analysis of deuterated cotton fibers under N 2 and air atmosphere were both altered due to the thermodynamic isotope effect. These observations revealed the hydrogen–deuterium exchange treatment process and impacts on cellulose fiber properties, which helped us to better understand the cellulose internal structure and may facilitate the potential utilization of deuterated cellulosic materials.
Paper sludge biomass represents an underutilized feedstock rich in pulped and processed cellulose which is currently a waste stream with significant disposal cost to industry for landfilling services. Effective fractionation of the cellulose from paper sludge presents an opportunity to yield cellulose as feedstock for value-added processes. A novel approach to cellulose fractionation is the sidehill screening system, herein studied at the pilot-plant scale. Composition analysis determined ash removal and carbohydrate retention of both sidehill and high-performance benchtop screening systems. Sidehill screening resulted in greater carbohydrates retention relative to benchtop screening (90% vs 66%) and similar ash removal (95% vs 98%). Techno-economic analysis for production of sugar syrup yielded a minimum selling price of $331/metric ton of sugar syrup including disposal savings, significantly less than a commercial sugar syrup without fractionation. Furthermore, sensitivity analysis showed that screening conditions played a significant role in economic feasibility for cellulosic yield and downstream processes.
Plant secondary cell walls constitute the dominant reservoir of renewable biomass, comprising tightly packed cellulose, hemicellulose, and lignin at the nanoscale. Recent advances in solid-state NMR spectroscopy and the availability of small-angle X-ray scattering for biomass characterization have led to an accumulation of experimental data on cell wall organization, yet no explicit structure model has simultaneously satisfied both Xray and NMR observations. Using wheat straw as a model system, we propose a structural framework consistent with current knowledge of cellulose biosynthesis, X-ray scattering data, and one- and two-dimensional 13 C solid-state NMR spectra. In this model, 18-chain elementary fibrils align in parallel and populate the cross-section at random. Arabinose-substituted xylan shows no conformational dependence for cellulose-binding in wheat, and only a minor fraction of 2-fold xylan appears in close proximity to cellulose, unlike in Arabidopsis, where xylan is more tightly attached to the cellulose surface. While NMR data cannot unambiguously resolve the internal arrangement of the 18 glucan chains, X-ray scattering profiles uniquely constrain the fibril size and exclude the possibility of tight bundling in the intact walls. The specific interaction between the matrix polymers and the cellulose elementary fibrils must be reconsidered in light of the small interfibril spaces, which bring the matrix components into spatial proximity with cellulose even in the absence of attractive interactions. These findings provide fundamental molecular-level insight into cellulose fibril architecture and matrix−polymer interactions, resolving longstanding discrepancies between spectroscopic and scattering data and advancing our understanding of biopolymer assembly into structurally and functionally versatile lignocellulosic biomaterials.
Plants synthesize polysaccharide-based primary cell walls that possess unique microstructures and mechanical properties to accommodate plant growth and provide protection. Here, it remains challenging to assess the role of polysaccharide organization and interactions in the mechanical behavior of primary cell walls owing to their complex microstructure and highly nonlinear mechanical responses. Employing a coarse-grained molecular dynamics model developed for onion epidermal walls, this work explores the conditions under which polysaccharide assembly and interactions might play a significant role in primary cell wall mechanics. Cellulose–cellulose adhesion plays a dominant role in the wall load-bearing capacity, but when cellulose–cellulose adhesion was disrupted computationally, cellulose–xyloglucan adhesion could influence the wall load-bearing capacity. Contrary to the common concept that xyloglucans mechanically tether well-separated cellulose microfibrils, xyloglucans functioned in this case as interfibrillar adhesives capable of transmitting tensile forces between cellulose microfibrils. Our findings may inform design criteria of new materials inspired by plant cell walls.