In silico structure prediction of full-length cotton cellulose synthase protein (GhCESA1) and its hierarchical complexes
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Microalgae are a promising feedstock for sustainable fuel and nutritional products. To be economically viable, algal biorefineries will need to produce multiple products, such as nutraceuticals and animal feed, in addition to fuel. Life cycle assessment (LCA) has been applied widely to algal biorefineries, but results vary across studies. Here we address two methodological challenges that can lead to variability in LCAs of multi-product algal biorefineries: the choice of functional unit and the method used to handle co-products. LCA was conducted for an algal biorefinery that produces two representative products—fuel and animal feed—using three functional units: (1) 1 MJ fuel, (2) 1 kg animal feed, and (3) 1 ha of production area, coupled with the system expansion method. For the fuel-based functional unit, the system expansion method was compared to impact allocation based on mass, energy, and market value of the biorefinery products. The choice of functional unit affects the results for several environmental indicators. For ecosystem quality, the choice of functional unit can change the overall balance of environmental impacts from harmful to beneficial. The effect of functional unit on climate change depends on the boundary considered in the model—i.e., “well to gate” or “well to wheels”. When a fuel-based functional unit is used, the choice of co-product handling method (i.e., allocation versus the system expansion method) affects environmental impacts, such as ecosystem quality and climate change, differently. For LCAs that use a fuel-based functional unit to compare algal biofuel to other fuels, multiple co-product handling methods should be analyzed and presented to stakeholders. A land based functional unit coupled with the system expansion method can be used to improve LCAs that compare among multi-product algal biorefineries.
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The hydrothermal liquefaction aqueous phase (HTL-AP), a nutrient- and carbon-rich waste byproduct of the HTL process, can be recirculated to enhance biocrude production efficiency. To avoid dilution of the HTL feedstock and reduced fuel quality associated with recirculating untreated HTL-AP, this study investigated nanofiltration to concentrate organic compounds and separate nutrients from HTL-AP of three algal biomass sources: an algal wastewater treatment system and two from harmful algal blooms in lakes. Up to 99% of the organics were recovered in the concentrated HTL-AP (retentate), and significant increases in both permeate flux and permeation of nitrogenous compounds were achieved by increasing HTL-AP temperature to 45 °C and pH to 11. Recirculating the concentrated HTL-AP increased biocrude yield by 70%-116% and reduced biocrude nitrogen content by up to 12%. The carbon and energy capture in the biocrude also increased by 66% and 68%, respectively.
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Plant cell walls are complex systems that exhibit the characteristics of both rigid and soft material depending on their external perturbations. The three main polymeric components in a plant primary cell wall are cellulose fibrils, hemicellulose, and pectins. These components interact in a hierarchical fashion giving rise to mesoscale structural features such as cellulose bundles, lamella stacking, and so on. Although several studies have focused on understanding these unique structural features, a clear picture linking them to cell wall mechanics is still lacking. As a first step toward this goal, a phenomenological model of plant cell wall has been developed in this work by using available experimental data to investigate the underlying connections between mesoscale structural features and the motions of fibrils during deformation. In this model cellulose fibrils exhibit motions such as angular reorientations and kinking upon forced stretching. Furthermore, these motions are dependent on the orientation of fibrils with respect to the stretch direction, i.e., fibrils that are at an angle to the stretch direction exhibit predominant angular reorientations, while fibrils transverse to the stretch direction undergo kinking as a result of transverse compression. Varying the chain length of pectin had negligible effects on these motions. One of the main contributions from this work is the development of a simple model that can be easily fine-tuned to test other hypotheses and extended to include additional experimental knowledge about the structural aspects of cell walls in the future.
Green pathways for nonisocyanate polyurethane (NIPU) production have attracted increasing levels of interest. The reaction between 5-membered cyclic carbonate and polyamines is one of the most promising pathways to produce NIPU polymers. Though promising, major technical hurdles such as slow polymerization rate and poor performance hinder the commercialization of NIPU. In this paper, we screened several commercially available triglyceride oil feedstocks for NIPU products, focusing on polymerization kinetics and product performance for industrial application. The impact of carbonated group density on polymerization rate and mechanical strength was determined. We have demonstrated a remarkably higher reactivity of carbonated oil derived from feedstocks with polyunsaturated fatty acid (PUFA). The NIPU derived from such feedstocks also showed improved performance for industrial application. Unlike traditional polyurethane foam production that uses isocyanate and water to generate CO2 as a blowing reagent, there is no gas formation in NIPU polymerization. We have demonstrated a practical and cost-effective approach to produce NIPU foam material using bicarbonate as a blowing reagent. Furthermore, we conducted the first-ever technoeconomic analysis (TEA), revealing that profitable commercial NIPU production can be achieved when operating at sufficient production capacities.
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Multi-response maps show that atomically dispersed iron on alumina provides relatively high activity and good stability in low temperature CO oxidation.
Solid-state nuclear magnetic resonance (ssNMR) is an indispensable tool for elucidating the structure and dynamics of insoluble and non-crystalline biomolecules. The recent advances in the sensitivity-enhancing technique magic-angle spinning dynamic nuclear polarization (MAS-DNP) have substantially expanded the territory of ssNMR investigations and enabled the detection of polymer interfaces in a cellular environment. This article highlights the emerging MAS-DNP approaches and their applications to the analysis of biomolecular composites and intact cells to determine the folding pathway and ligand binding of proteins, the structural polymorphism of low-populated biopolymers, as well as the physical interactions between carbohydrates, proteins, and lignin. Furthermore, these structural features provide an atomic-level understanding of many cellular processes, promoting the development of better biomaterials and inhibitors. It is anticipated that the capabilities of MAS-DNP in biomolecular and biomaterial research will be further enlarged by the rapid development of instrumentation and methodology.
De-esterification of homogalacturonan (HG) is thought to stiffen pectin gels and primary cell walls by increasing calcium cross-linking between HG chains. Contrary to this idea, recent studies found that HG de-esterification correlated with reduced stiffness of living tissues, measured by surface indentation. The physical basis of such apparent wall softening is unclear, but possibly involves complex biological responses to HG modification. To assess the direct physical consequences of HG de-esterification on wall mechanics without such complications, we treated isolated onion (Allium cepa) epidermal walls with pectin methylesterase (PME) and assessed wall biomechanics with indentation and tensile tests. In nanoindentation assays, PME action softened the wall (reduced the indentation modulus). In tensile force/extension assays, PME increased plasticity, but not elasticity. These softening effects are attributed, at least in part, to increased electrostatic repulsion and swelling of the wall after PME treatment. Despite softening and swelling upon HG de-esterification, PME treatment alone failed to induce cell wall creep. Instead, acid-induced creep, mediated by endogenous α-expansin, was reduced. We conclude that HG de-esterification physically softens the onion wall, yet reduces expansin-mediated wall extensibility.
Auxin-induced cell elongation relies in part on the acidification of the cell wall, a process known as acid growth that presumably triggers expansin-mediated wall loosening via altered interactions between cellulose microfibrils. Cellulose microfibrils are a major determinant for anisotropic growth and they provide the scaffold for cell wall assembly. Little is known about how acid growth depends on cell wall architecture. To explore the relationship between acid growth-mediated cell elongation and plant cell wall architecture, two mutants (jia1-1 and csi1-3) that are defective in cellulose biosynthesis and cellulose microfibril organization were analyzed. The study revealed that cell elongation is dependent on CSI1-mediated cell wall architecture but not on the overall crystalline cellulose content. We observed a correlation between loss of crossed-polylamellate walls and loss of auxin- and fusicoccin-induced cell growth in csi1-3. Furthermore, induced loss of crossed-polylamellate walls via disruption of cortical microtubules mimics the effect of csi1 in acid growth. We hypothesize that CSI1- and microtubule-dependent crossed-polylamellate walls are required for acid growth in Arabidopsis hypocotyls.
Genome assembly and annotation of the biofuel- and bioproduct-relevant microalga Nannochloropsis oceanica CCAP849/10
Plant cell walls are dynamic structures that are synthesized by plants to provide durable coverings for the delicate cells they encase. They are made of polysaccharides, proteins, and other biomolecules and have evolved to withstand large amounts of physical force and to resist external attack by herbivores and pathogens but can in many cases expand, contract, and undergo controlled degradation and reconstruction to facilitate developmental transitions and regulate plant physiology and reproduction. Recent advances in genetics, microscopy, biochemistry, structural biology, and physical characterization methods have revealed a diverse set of mechanisms by which plant cells dynamically monitor and regulate the composition and architecture of their cell walls, but much remains to be discovered about how the nanoscale assembly of these remarkable structures underpins the majestic forms and vital ecological functions achieved by plants.
Microbial lipids hold great promise as biofuel precursors, and research efforts to convert such lipids to renewable diesel fuels have been increasing in recent years. In contrast to the numerous literature reviews on growing, characterising and extracting lipids from oleaginous microbes, and on converting vegetable oils to hydrocarbon fuels, this review aims to provide insight into aspects that are specific to hydroprocessing microbial lipids. While standard hydrotreating catalysts generally perform well with terrestrial oils, differences in lipid speciation and the presence of co-extracted compounds, such as chlorophyll and sterols, introduce additional complexities into the process for microbial lipids. Lipid cleanup steps can be introduced to produce suitable feedstocks for catalytic upgrading.
Algenol Biotech, the National Renewable Energy Laboratory (NREL), Georgia Institute of Technology, Arizona State University, and Reliance Industries formed a team to advance the state of the art in algal biomass production and downstream processing technologies, with the end goal of a sustainable, economically viable biofuel intermediate (BFI, biocrude) product. The project included examination of high value co-product production as a market entry strategy and for enhancing the economics of a biorefinery for BFI production. The project targeted innovations in biology, operations, and engineering. The goals of the project were: BFI productivity greater than 4,000 gal-BFI/acre-yr on an annualized basis; energy efficient innovations in downstream operations resulting in an energy expenditure less than 10% of the BFI energy content and a carbon footprint reduction of more than 60% compared to fossil alternatives; and a comprehensive Techno-Economic Analysis (TEA) that identifies limiting factors for commercial viability of a photobioreactor (PBR)-based biofuel product. The project achieved the overall objectives. Strain development efforts led to the identification of a strain (Cyanobacterium sp. AB1166) that, relative to the previous best strain (Cyanobacterium sp. AB1), exhibited a ~10% increase in productivity under commercially-relevant cultivation conditions and also resulted in cultures with a >50% reduction in viscosity such that harvesting efficiency was improved; these results represent achievement of key project milestones. Progress was also made at NREL in altering the biochemical composition of algal biomass to improve the yield of BFI produced via HTL. These strain enhancements coupled with improved outdoor cultivation practices, including semi-continuous operation, increased areal biomass productivity by nearly 80% over the established baseline productivity. The annualized productivity achieved (26.8 g/m2-d), paired with HTL conversion yields realized at NREL and RIL (38% ± 2% BFI), translates to 4,100 gal-BFI/acre-yr, exceeding the FY20 BETO goal of 3,700 gal-BFI/acre-yr. Significant progress was also demonstrated in large scale PBR-based production system design, operability, and cost reduction. A 24,000-L production module comprised of 240 interlinked PBRs was constructed and successfully operated outdoors for over one year in Fort Myers, Florida. Aided by a state-of-the-art productivity model, the productivities achieved convincingly demonstrated scalability of laboratory results determined at the mL to L scale to large-scale outdoor operations exceeding 20,000 L. The system was used to cultivate Arthrospira platensis (Spirulina), an industrially-relevant cyanobacterium and source for phycocyanin, an approved blue food colorant that Algenol is developing as a risk reduction strategy for future biofuel projects and as a potential business opportunity. A key project milestone to develop phycocyanin extraction and purification technologies was achieved ahead of schedule, and product samples received positive feedback from potential customers. The production and downstream operations data generated in this project were used to conduct and refine Techno-Economic and Life Cycle Assessments to provide research guidance for reducing the costs and environmental footprint of algal biofuel and co-product manufacturing plants. Several CO 2 supply scenarios for an algal biorefinery were identified as being capable of providing a large (>60%) reduction in carbon footprint in comparison to gasoline. The TEA assessments incorporated detailed comparisons of PBR versus open pond production systems, yielding a 3-fold higher areal productivity for PBRs and suggesting overall production cost parity for the two systems. The progress in this ABY2 project addressed many of the the barriers identified for the Advanced Algal Systems R&D Program and are directly relevant to achieving the established BETO goals associated with large scale biofuel production and cost reduction.
The report documents the conceptual basis for a new potential Combined Algal Processing design strategy which may allow more flexibility in accommodating different algal biomass feedstock compositions, by enabling upgrading of both protein and carbohydrates in a single step, without a strict requirement for either component to be in soluble or monomeric form, while maintaining effective wet lipid extraction techniques to enable high lipid recoveries. In light of previously-established constraints around algal biomass costs (which are significantly higher than lignocellulosic terrestrial biomass), the present CAP processing strategy reflects an integrated biorefinery concept producing both fuels and value-added chemical coproducts as a means to improve profitability and generate coproduct revenues to help drive down the minimum fuel selling price (MFSP) towards economically viable levels. Namely, this report highlights an integrated CAP biorefinery process and associated technical targets that would be required to achieve U.S. Department of Energy target MFSP goals of $2.5/gallon gasoline equivalent by 2030. This is accomplished by a process involving low-cost seasonal storage of algal biomass during high-growth seasons, rapid flash hydrolysis pretreatment of the biomass, solvent extraction of pretreated biomass, cleanup and fractionation of lipids into triglyceride and free fatty acid fractions, and a series of thermochemical conversion steps to upgrade carbohydrates and protein to hydrocarbon fuels. These steps include mild oxidative treatment (MOT), a process originally investigated at NREL for upgrading lignin, followed by catalytic ketonization and hydrotreating of MOT products to fuels. Isolated triglycerides are sent to a coproduct train, with the base case focused on upgrading to polyurethane foams as a high-value, high-market-volume coproduct.