Bioconversion of cellulose into bisabolene using Ruminococcus flavefaciens and Rhodosporidium toruloides
Not Available
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SEARCH · Engineering Papers
Results for “Cellulosic biofuel”
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Lignin-first biorefining of Nordic poplar to produce cellulose fibers could displace cotton production on agricultural lands
Here, we show that lignin-first biorefining of poplar can enable the production of dissolving cellulose pulp that can produce regenerated cellulose, which could substitute cotton. These results in turn indicate that agricultural land dedicated to cotton could be reclaimed for food production by extending poplar plantations to produce textile fibers. Based on climate-adapted poplar clones capable of growth on marginal lands in the Nordic region, we estimate an environmentally sustainable annual biomass production of ~11 tonnes/ha. At scale, lignin-first biorefining of this poplar could annually generate 2.4 tonnes/ha of dissolving pulp for textiles and 1.1 m 3 biofuels. Life cycle assessment indicates that, relative to cotton production, this approach could substantially reduce water consumption and identifies certain areas for further improvement. Overall, this work highlights a new value chain to reduce the environmental footprint of textiles, chemicals, and biofuels while enabling land reclamation and water savings from cotton back to food production.
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Inhibition of enzyme hydrolysis of cellulose by phenols from hydrothermally pretreated sugarcane straw
Relatively few studies have addressed the characterization of sugarcane straw (SCS) for production of fermentable sugars through enzyme hydrolysis. Straw is a major co-product of the sugarcane harvest in Brazil that has potential to sustainably increase cellulosic feedstocks in Brazil by 50%. Pretreatment of 10% w/v straw with liquid hot water (LHW) at 180°C for 50 min (severity, S of 4.05), solubilizes hemicellulose, preserves glucan, and generates 4.49 g/L soluble phenolic compounds in the resulting liquid. Extracts from washing pretreated solids with excess hot water followed by acetone resulted in 1.10 and 0.83 g/L phenolics, respectively. Here, acetone-derived extracts were more inhibitory and decreased glucose yield for enzyme hydrolysis of Solka Floc (a lignin-free cellulose) by 42%. In comparison, pretreated straw washed with hot water or acetone was readily hydrolyzed to 92% and 97% by cellulase enzyme. Hydrothermally treated SCS has the potential to provide a valuable and added source of fermentable sugars suitable for bioprocessing into biofuels and bioproducts when cellulase enzyme inhibitors are removed after pretreatment.
Deactivation↗
Editorial: Lignocellulose valorization: Fractionation, conversion and applications
Lignocellulosic biomass is the most abundant form of renewable feedstock on the Earth. Chemicals, energies, and materials derived from lignocellulosic biomass are renewable and sustainable, and have the potential to replace fossil feedstocks. Biorefinery, a process to fractionate lignocellulose into the three major components, is considered the most pivotal step for bioenergy, biomaterials and biochemicals. However, due to the complex hierarchy and chemical structures, only limited lignocellulose is valorized into value-added products, and most of them are burnt or just discarded. Therefore, there is a need to investigate the heterogeneous structure of lignocellulose, improve its efficient fractionation into cellulose, hemicelluloses and lignin, and develop the technology that can convert the lignocellulose into high-value chemical products and high-performance functional materials in a sustainable and promising configuration. In short, the ultimate goal of biomass utilization is that the production of biofuels and biomaterials in industrial applications must be cost-, and performance-competitive with petroleum-derived equivalents.
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
Editorial: Wood Development and Physiology in a Changing Climate
Wood of forest trees is a renewable, sustainable and easily workable material and has been widely used in construction, paper making, furniture, and as a feedstock for biofuels. Wood formation is the result of a highly ordered and finely regulated process. Secondary xylem (wood) and phloem are the inner and outer derivative products of the vascular cambium, respectively. Xylem is mainly composed of dead cells with thickened cell walls rich in cellulose, hemicelluloses, and lignin, which are responsible for providing mechanical support and conducting water and minerals for the trees. Phloem transports photoassimilates and signaling molecules, including phytohormones and peptides. Wood structure varies greatly between, and within, tree species. A thorough understanding of developmental processes and metabolic pathways during wood formation would provide a basis for improving wood biomass and modifying properties in forest trees.
54 ENVIRONMENTAL SCIENCES↗
Field evaluation of transgenic hybrid poplars with desirable wood properties and enhanced growth for biofuel production by bicistronic expression of PdGA20ox1 and PtrMYB3 in wood-forming tissue
Background: To create an ideotype woody bioenergy crop with desirable growth and biomass properties, we utilized the viral 2A-meidated bicistronic expression strategy to express both PtrMYB3 ( MYB46 ortholog of Populus trichocarpa , a master regulator of secondary wall biosynthesis) and PdGA20ox1 (a GA20-oxidase from Pinus densiflora that produces gibberellins) in wood-forming tissue (i.e., developing xylem). Results: Transgenic Arabidopsis plants expressing the gene construct DX15::PdGA20ox1-2A-PtrMYB3 showed a significant increase in both stem fresh weight (threefold) and secondary wall thickening (1.27-fold) relative to wild-type (WT) plants. Transgenic poplars harboring the same gene construct grown in a greenhouse for 60 days had a stem fresh weight up to 2.6-fold greater than that of WT plants. In a living modified organism (LMO) field test conducted for 3 months of active growing season, the stem height and diameter growth of the transgenic poplars were 1.7- and 1.6-fold higher than those of WT plants, respectively, with minimal adverse growth defects. Although no significant changes in secondary wall thickening of the stem tissue of the transgenic poplars were observed, cellulose content was increased up to 14.4 wt% compared to WT, resulting in improved saccharification efficiency of the transgenic poplars. Moreover, enhanced woody biomass production by the transgenic poplars was further validated by re-planting in the same LMO field for additional two growing seasons. Conclusions: Taken together, these results show considerably enhanced wood formation of our transgenic poplars, with improved wood quality for biofuel production.
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Transgenic Poplar Designed for Biofuels
Members of the genus Populus (i.e., cottonwood, hybrid poplar) represent a promising source of lignocellulosic biomass for biofuels. However, one of the major factors negatively affecting poplar’s efficient conversion to biofuel is the inherent recalcitrance to enzymatic saccharification due to cell wall components such as lignin. To this effect, there have been efforts to modify gene expression to reduce biomass recalcitrance by changing cell wall properties. Furthermore, we review recent genetic modifications of poplar that led to change cell wall properties and the resulting effects on subsequent pretreatment efficacy and saccharification. Although genetic engineering’s impacts on cell wall properties are not fully predictable, recent studies have shown promising improvement in the biological conversion of transgenic poplar to biofuels.
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In silico structure prediction of full-length cotton cellulose synthase protein (GhCESA1) and its hierarchical complexes
Not provided.
biofuels (including algae and biomass), bio-inspir↗
Feedstock Variability: Differences Among Biomass Anatomical Fractions and Tissues
Corn stover is a versatile feedstock for producing biofuels and other sustainable products. However, its diverse chemical and physical attributes can hinder the processes that convert it into these compounds. The composition of corn stover varies according to factors including corn plant variety, cultivation conditions and maturity at harvest. In essence, corn stover consists of the polymers cellulose, hemicellulose and lignin. The structural core of plant cell walls consists of elongated chains of glucose molecules arranged in bundles of cellulose. Additionally, lower concentrations of hemicellulose, a complex carbohydrate composed of various sugars, form part of the matrix surrounding cellulose bundles. Lignin is a complex aromatic polymer that reinforces plant cell walls. Corn stover generally comprises 40%-50% cellulose, 20%-30% hemicellulose, and 20%-30% lignin. The ratios of these components vary based on the specific type of corn plant and the growth conditions. For example, corn plants cultivated in drought conditions exhibit higher lignin levels than those grown under optimal conditions. The compositional diversity resulting from growth conditions is most pronounced among the different anatomical fractions of the corn plant. Understanding and accommodating the variation among the anatomical fractions of corn stover can enhance the effective utilization of this resource.
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THF co-solvent pretreatment prevents lignin redeposition from interfering with enzymes yielding prolonged cellulase activity
Background: Conventional aqueous dilute sulfuric acid (DSA) pretreatment of lignocellulosic biomass facilitates hemicellulose solubilization and can improve subsequent enzymatic digestibility of cellulose to fermentable glucose. However, much of the lignin after DSA pretreatment either remains intact within the cell wall or readily redeposits back onto the biomass surface. This redeposited lignin has been shown to reduce enzyme activity and contribute to rapid enzyme deactivation, thus, necessitating significantly higher enzyme loadings than deemed economical for biofuel production from biomass. Results: In this study, we demonstrate how detrimental lignin redeposition on biomass surface after pretreatment can be prevented by employing Co-solvent Enhanced Lignocellulosic Fractionation (CELF) pretreatment that uses THF–water co-solvents with dilute sulfuric acid to solubilize lignin and overcome limitations of DSA pretreatment. We first find that enzymatic hydrolysis of CELF-pretreated switchgrass can sustain a high enzyme activity over incubation periods as long as 5 weeks with enzyme doses as low as 2 mg protein/g glucan to achieve 90% yield to glucose. A modified Ninhydrin-based protein assay revealed that the free-enzyme concentration in the hydrolysate liquor, related to enzyme activity, remained unchanged over long hydrolysis times. DSA-pretreated switchgrass, by contrast, had a 40% drop in free enzymes in solution during incubation, providing evidence of enzyme deactivation. Furthermore, measurements of enzyme adsorption per gram of lignin suggested that CELF prevented lignin redeposition onto the biomass surface, and the little lignin left in the solids was mostly integral to the original lignin–carbohydrate complex (LCC). Scanning electron micrographs and NMR characterization of lignin supported this observation. Conclusions: Enzymatic hydrolysis of solids from CELF pretreatment of switchgrass at low enzyme loadings was sustained for considerably longer times and reached higher conversions than for DSA solids. Analysis of solids following pretreatment and enzymatic hydrolysis showed that prolonged cellulase activity could be attributed to the limited lignin redeposition on the biomass surface making more enzymes available for hydrolysis of more accessible glucan.
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Biomass formation and sugar release efficiency of Populus modified by altered expression of a NAC transcription factor
Woody biomass is an important feedstock for biofuel production. Manipulation of wood properties that enable efficient conversion of biomass to biofuel reduces cost of biofuel production. Wood cell wall composition is regulated at several levels that involve expression of transcription factors such as wood-/secondary cell wall-associated NAC domains (WND or SND). In Arabidopsis thaliana, SND1 regulates cell wall composition through activation of its down-stream targets such as MYBs. The functional aspects of SND1 homologs in the woody Populus have been studied through transgenic manipulation. In this study, we investigated the role of PdWND1B, Populus SND1 sequence ortholog, in wood formation using transgenic manipulation through over-expression or silencing under the control of a vascular-specific 4-coumarate-CoA ligase (4CL) promoter. As compared with control plants, PdWND1B-RNAi plants were shorter in height, with significantly reduced stem diameter and dry biomass, whereas there were no significant differences in growth and productivity of PdWND1B over-expression plants. Conversely, PdWND1B over-expression lines showed a significant reduction in cellulose and increase in lignin content, whereas there was no significant impact on lignin content of downregulated lines. Stem carbohydrate composition analysis revealed a decrease in glucose, mannose, arabinose, and galactose, but an increase in xylose in the over-expression lines. Transcriptome analysis revealed upregulation of several downstream transcription factors and secondary cell wall related structural genes in the PdWND1B over-expression lines, partly explaining the observed phenotypic changes in cell wall chemistry. Relative to the control, glucose release efficiency and ethanol production from stem biomass was significantly reduced in over-expression lines. Our results show that PdWND1B is an important factor determining biomass productivity, cell wall chemistry and its conversion to biofuels in Populus.
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Agroecosystem model simulations reveal spatial variability in relative productivity in biomass sorghum and maize in Iowa, USA
Biomass sorghum (Sorghum bicolor L. Moench) is an annual C 4 grass that has emerged as a candidate bioenergy crop but has not been widely grown in the United States. Corn (Zea mays L.), another annual C 4 grass, has been produced on a large scale in the United States for biofuels. Iowa leads the nation in both corn and ethanol production. The high productivity of corn in Iowa creates the research question: could biomass sorghum be as or more productive than corn in the state in terms of bioenergy? Efforts to use crop models to fill the gaps left by field experimentation on biomass sorghum have also been limited thus far. To address our research question, we collected biophysical data on biomass sorghum grown in Iowa for two growing seasons, used them to develop a biomass sorghum module in an agroecosystem model (Agro-IBIS), and predict the potential performance of biomass sorghum across Iowa compared to maize. Despite dry conditions in 2019 and 2020, average biomass sorghum yields were 17.20 Mg ha –1 . By comparison, average corn aboveground biomass was higher in 2020 (22.02 Mg ha –1 ). Soil cores indicated average belowground biomass of 1.46 Mg ha –1 , with roots concentrated near the surface (73% of biomass above 50 cm in a 1 m core). When biomass sorghum model parameters were calibrated with measured values, model output was in close agreement with measured biomass (slope = 0.932, R 2 = 0.91) and evapotranspiration (slope = 0.757, R 2 = 0.64). Subsequent regional simulations revealed a notable latitudinal gradient in biomass sorghum yield, with a strong linear relationship between yield and seasonal growing degree-days (R 2 = 0.89). When these yields were compared to simulated corn aboveground biomass yields, only 3.4% of the state had biomass sorghum yields that were significantly higher than corn. This number was reduced to 0.3% when comparing the crops in terms of energy ethanol yield. Thus, we conclude biomass sorghum must be improved to be competitive with corn as an annual biofuel crop in Iowa. This study provides a baseline against which to compare advancements in sorghum breeding for biomass and stress tolerance in Iowa.
Agro-IBIS↗
Yield, chemistry, and TEA/LCA data for 84 switchgrass (Panicum virgatum L.) variants
Switchgrass yield, chemistry, technoeconomic assessment (TEA), and life cycle assessment (LCA) data referenced in the manuscript "Economics and Sustainability impacts of yield and composition variation in bioenergy crops: Panicum virgatum L.", Happs, et. al. (manuscript in revision at ACS Sustainable Chemistry & Engineering). These results are based on analysis of yield data generated from a set of switchgrass common garden experiments established by the Center for Bioenergy Innovation (CBI; https://cbi.ornl.gov/). These data include estimates of commercial-scale yields and characterization of biomass chemistry (fermentable carbohydrate fraction) for 84 different switchgrass variants from a genome-wide association study population, as well as estimates of minimum fuel selling price and various life-cycle footprint metrics for cellulosic ethanol produced from that switchgrass biomass.
switchgrass, biofuels↗
Can a Nuclear-Assisted Biofuels System Enable Liquid Biofuels as the Economic Low-carbon Replacement for All Liquid Fossil Fuels and Hydrocarbon Feedstocks and Enable Negative Carbon Emissions?
This report integrates the results of a series of studies and workshops to address the question: Can a nuclear-assisted biofuels system enable liquid biofuels as the economic low-carbon replacement for all liquid fossil fuels and hydrocarbon feedstocks and simultaneously enable negative carbon emissions? “Economic” is defined as economically competitive relative to other low-carbon replacement options for crude oil. “All” refers to the capability to replace the 18 million barrels of oil per day used by the United States. “Nuclear-assisted” refers to the provision of massive quantities of low-carbon heat and hydrogen at the refinery to fully utilize the carbon content of the cellulosic biomass feedstocks for liquid hydrocarbon production. A system is proposed that decarbonizes about half of the U.S. economy while improving long-term soil productivity and sequestering carbon from the atmosphere. In the U.S. almost half the energy consumed by the final customer is in the form of liquid hydrocarbons produced from crude oil. Liquid hydrocarbons are used as (1) an energy source, (2) a method for daily-to-seasonal energy storage, (3) a chemical feedstock, (4) a chemical reducing agent, (5) a method to enhance high-temperature heat transfer in many furnaces and industrial processes and (6) other purposes. As a consequence, the U.S. consumes ~18 million barrels of crude oil per day to produce liquid hydrocarbons. While there are substitutes for liquid hydrocarbons for some applications, our assessment is that the costs and difficulty will dramatically increase if liquid hydrocarbon use goes much below the equivalent of 10 million barrels per day of crude oil. New uses of liquid hydrocarbons to partly replace coal and natural gas could increase demand beyond the equivalent of 10 million barrels of oil per day. One can produce and burn liquid hydrocarbons from biomass without any net addition of carbon dioxide to the atmosphere. Plants grow by removing carbon dioxide from the atmosphere; thus, burning biomass does not result in any net addition of carbon dioxide to the atmosphere. Biomass is typically 40% oxygen. To remove this oxygen to create hydrocarbon liquids, there are two options. The first option is to use biomass as (1) a feedstock, (2) an energy source to operate the process and (3) a supply of carbon to remove the biomass oxygen as carbon dioxide. The second option is to use external heat and hydrogen to remove the oxygen as water and produce liquid hydrocarbons. The use of massive quantities of external heat and hydrogen for hydrocarbon liquid fuels production reduces the biomass feedstock per unit of liquid hydrocarbon product by more than a factor of two reducing land use by more than a factor of two. Many cellulosic feedstocks unsuitable for liquid hydrocarbon production are viable feedstocks with external heat and hydrogen inputs. As a result, there is sufficient cellulosic feedstocks to meet U.S. and global liquid fuels hydrocarbon demand without significant impacts on food and fiber prices. The heat and hydrogen (heat plus electricity) are produced using base-load nuclear power plants—the most economic form of nuclear energy. If low-price natural gas, there is also the option of producing hydrogen from natural gas with sequestration of the carbon dioxide. The biomass is locally processed in depots to produce commodity feedstocks that can be shipped long distances to large biorefineries (250,000 barrels per day, oil equivalent) similar to existing refineries except modifications of front-end processes. The depot system enables local recycle of nutrients back to the soil. The biorefinery can produce variable quantities of liquid hydrocarbon fuels and carbon dioxide for sequestration enabling removal of carbon dioxide from the atmosphere. Preliminary estimates are that the liquid hydrocarbon costs are equivalent to crude oil at between $60 and $70 per barrel. The largest cost is for hydrogen with biomass feedstocks and refinery costs a smaller fraction of the total cost.
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A grass-specific cellulose–xylan interaction dominates in sorghum secondary cell walls
Abstract Sorghum ( Sorghum bicolor L. Moench) is a promising source of lignocellulosic biomass for the production of renewable fuels and chemicals, as well as for forage. Understanding secondary cell wall architecture is key to understanding recalcitrance i.e. identifying features which prevent the efficient conversion of complex biomass to simple carbon units. Here, we use multi-dimensional magic angle spinning solid-state NMR to characterize the sorghum secondary cell wall. We show that xylan is mainly in a three-fold screw conformation due to dense arabinosyl substitutions, with close proximity to cellulose. We also show that sorghum secondary cell walls present a high ratio of amorphous to crystalline cellulose as compared to dicots. We propose a model of sorghum cell wall architecture which is dominated by interactions between three-fold screw xylan and amorphous cellulose. This work will aid the design of low-recalcitrance biomass crops, a requirement for a sustainable bioeconomy.
59 BASIC BIOLOGICAL SCIENCES↗
Environmental Trade-Offs for a Process Utilizing Aerobic Respiration for Biological Conversion of Cellulosic Biomass-Derived Sugars to Hydrocarbons (Renewable Diesel Blendstock): Cooperative Research and Development Final Report, CRADA Number CRD-18-00765, Project 1
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.
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Estimating induced land use change emissions for sustainable aviation biofuel pathways
Sustainable aviation fuels (SAFs) are expected to play an essential role in achieving the aviation industries’ goal of carbon-neutral growth. However, producing biomass-based SAFs may induce changes in global land use and the associated carbon stock. The induced land use change (ILUC) emissions, as a part of the full life-cycle emissions for SAF pathways, will affect whether and to what extent SAFs reduce emissions compared with petroleum-based jet fuels. Here, we estimate the ILUC emission intensity for seventeen SAF pathways considered by the International Civil Aviation Organization (ICAO), covering five ASTM-certified technologies, nine biomass-based feedstocks, and four geographical regions. We introduce the SAF pathways into a well-established computable general equilibrium (CGE) model, GTAP-BIO, and its coupled emission accounting model, AEZ-EF, to study economy-wide implications of SAF production and estimate ILUC emissions intensity for each pathway. The estimated SAF ILUC emission intensities, using a 25-year amortization period, range from -58.5 g CO 2 e MJ -1 for the USA miscanthus alcohol (isobutanol)-to-jet (ATJ) pathway to 34.6 g CO 2 e MJ -1 for the Malaysia & Indonesia palm oil Hydrotreated Esters of Fatty Acids (HEFA) pathway. Notably, the vegetable oil pathways tend to have higher ILUC emission intensities due to their linkage to palm expansion and peatland oxidation in Southeast Asia. The cellulosic pathways studied provide negative ILUC emissions, mainly driven by the high carbon sequestrations in crop biomass and soil. Using the core life-cycle emissions established by ICAO, we show that fifteen of the assessed pathways have a lower full life-cycle emission intensity than petroleum-based jet fuels (89 g CO 2 e MJ -1 ), offering promising options to reduce aviation emissions.
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Acoustic force spectroscopy reveals subtle differences in cellulose unbinding behavior of carbohydrate-binding modules
Protein adsorption to solid carbohydrate interfaces is critical to many biological processes, particularly in biomass deconstruction. To engineer more-efficient enzymes for biomass deconstruction into sugars, it is necessary to characterize the complex protein–carbohydrate interfacial interactions. A carbohydrate-binding module (CBM) is often associated with microbial surface-tethered cellulosomes or secreted cellulase enzymes to enhance substrate accessibility. However, it is not well known how CBMs recognize, bind, and dissociate from polysaccharides to facilitate efficient cellulolytic activity, due to the lack of mechanistic understanding and a suitable toolkit to study CBM–substrate interactions. Our work outlines a general approach to study the unbinding behavior of CBMs from polysaccharide surfaces using a highly multiplexed single-molecule force spectroscopy assay. Here, we apply acoustic force spectroscopy (AFS) to probe a Clostridium thermocellum cellulosomal scaffoldin protein (CBM3a) and measure its dissociation from nanocellulose surfaces at physiologically relevant, low force loading rates. An automated microfluidic setup and method for uniform deposition of insoluble polysaccharides on the AFS chip surfaces are demonstrated. The rupture forces of wild-type CBM3a, and its Y67A mutant, unbinding from nanocellulose surfaces suggests distinct multimodal CBM binding conformations, with structural mechanisms further explored using molecular dynamics simulations. Applying classical dynamic force spectroscopy theory, the single-molecule unbinding rate at zero force is extrapolated and found to agree with bulk equilibrium unbinding rates estimated independently using quartz crystal microbalance with dissipation monitoring. However, our results also highlight critical limitations of applying classical theory to explain the highly multivalent binding interactions for cellulose–CBM bond rupture forces exceeding 15 pN.
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