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Scalable and Tunable Carbide-Phosphide Composite Catalyst System for the Thermochemical Conversion of Biomass

We have prepared composite materials of hexagonal nickel phosphide and molybdenum carbide (Mo2C) utilizing a simple and scalable two-stage synthesis method comprised of carbothermic reduction followed by hydrothermal incubation. We observe the monophasic hexagonal phosphide Ni2P in the composite at low phosphide-to-carbide (P:C) ratios. Upon increasing the proportion of P:C, the carbide surface becomes saturated, and we detect the emergence of a second hexagonal nickel phosphide phase (Ni5P4) upon annealing. We demonstrate that vapor-phase upgrading (VPU) of whole biomass via catalytic fast pyrolysis is achievable using the composite material as a catalyst, and we monitor the resulting product slates using pyrolysis gas chromatography/mass spectrometry. Our analysis of the product vapors indicates that variation of the P:C molar ratio in the composite material affords product slates of varying complexity and composition, which is indicated by the number of products and their relative proportions in the product slate. Our results demonstrate that targeted vapor product composition can be obtained, which can potentially be utilized to tune the composition of the bio-oil downstream.

09 BIOMASS FUELS↗

Evaluation of Fifteen Cultivars of Cool‐Season Perennial Grasses as Biofuel Feedstocks Using Near‐Infrared

Core Ideas Harvest yield varies more across species than sugar content and accessibility. Harvest yield and sugar accessibility are both critical parameters for conversion. Near‐infrared/partial least square models are valuable for quickly evaluating biomass for bioconversion. Cool‐season (C3) perennial grasses have a long history of cultivation and use as animal forage. This study evaluated 15 cultivars of C3 grasses, when harvested in late June for increased biomass yield, as biofuel feedstocks using near‐ infrared spectroscopy (NIR) based partial least square (PLS) analysis. These grasses were grown near Iliff, CO, for three growing seasons (2009–2011). The carbohydrate composition and released carbohydrates (total glucose and xylose released from dilute acid pretreatment and enzymatic hydrolysis [EH]) were predicted for samples from the study using NIR/PLS. The results were analyzed from a biofuels perspective, where composition combined with harvest yield provided information on the carbohydrate yield available for biomass conversion processes, and released carbohydrate yield provided information on the accessibility of those carbohydrates to conversion methods. The range in harvest yields varied more among cultivars (2900 kg ha −1 ) than did the range in carbohydrate composition (56.0 g kg −1 ) or released carbohydrates (60.0 g kg −1 ). When comparing carbohydrate yield to released carbohydrate yield between cultivars, an efficiency as high as 87% release of available carbohydrates was obtained for pubescent wheatgrass [ Thinopyrum intermedium (Host) Barkworth & D.R. Dewey ‘Mansaka’], with a low of 71% for hybrid wheatgrass [ Elytrigia repens (L.) nevski ´ pseudoroegneria spicata (PURSH) A. Love ‘Newhy’]. Though hybrid wheatgrass had the lowest release efficiency, its high harvest yield resulted in release of more total carbohydrates than half the other cultivars analyzed. This suggested that harvest yield, carbohydrate release, and carbohydrate composition, togetherplay significant roles in biofuel feedstock evaluation.

09 BIOMASS FUELS↗

Chapter 7.1: Microalgae

Microalgae is a unique biomass resource that does not need to compete for land and water with other biomass feedstocks because it can be cultivated on low-quality unencumbered land using noncompetitive water types including saline and wastewater. It is included as a complementary resource alongside other biomass feedstocks reported in this study, albeit at higher biomass production costs reflective of more capital-intensive farming operations than typical for terrestrial biomass. Higher biomass costs can be offset by the potential to produce value-added coproducts unique to compositional constituents of microalgae. Relative to BT16, this chapter reflects the latest analysis from the 2022 Algae Harmonization Update, which uses the latest parameterized and high-performing saline algal strain, second-generation carbon capture of point-source waste CO2 and high-pressure pipeline transport resolved to specific point-source types, saline water sourcing up to 40,000 mg/L total dissolved solids for source and makeup water salinity, blowdown water treatment and recycle, and brine disposal handling. National-scale algal biomass availability potential was calculated at 152 million tons/yr ash-free dry weight (AFDW) (191 million tons/yr dry weight) at an average biomass productivity of 26.2 g/m2 -day AFDW (about 50 tons/acre/yr dry weight).1 The algal biomass was cultivated on 3.9 million acres of multi-criteria screened and potentially available land for CONUS and fixed 268 million tons of waste CO2 based on biomass uptake. The algae biomass can be produced at an average MBSP of $674/ton AFDW ($536/ton dry weight) in 2020 dollars,2 corresponding to a total energy potential of 3.3 quads/yr at an average MBSP of $31.2/MMBtu (higher-heating-value [HHV] basis).

algae↗

Technology Case Study: Techno-Economic and Life Cycle Analysis for Microalgae Conversion Pathways to Fuels and Products

This technology case study report details the cost and sustainability prospects for an emerging feedstock - microalgae - converted to fuels and products via a fractionation and upgrading approach termed combined algae processing (CAP). Detailed techno-economic analysis (TEA) and life cycle analysis (LCA) are conducted for the conversion of farmed algae biomass, with two primary scenarios considering the conversion of either high-compositional-quality biomass enriched in lipids (high-lipid [HL]) or lower-quality biomass enriched in protein (high-protein [HP]). Each scenario employs a different biorefinery configuration tailored towards extracting the maximum value from the given biomass composition. The HL scenario produces fuels and non-isocyanate polyurethane (NIPU) as the primary products, while the HP scenario products fuels and a residual solid coproduct which can be used as a co-feed for producing thermoplastics. The results for the HL scenario were particularly promising, with a minimum fuel selling price (MFSP) of $\$$3.68 per gasoline gallon equivalent (GGE) and fuel GHG emissions translating to 54%-76% reduction compared to petroleum fuels depending on the coproduct handling method used. In contrast, the HP scenario faced more challenges in producing biofuels economically, projecting an MFSP of $\$$7.92/GGE despite significant revenues from the residual algae solids. LCA results for the HP case reflected a 24% reduction potential in biorefinery-level GHG emissions. However, these GHG reductions were primarily associated with the thermoplastic coproduct, which accounted for 93% of all biorefinery outputs by mass. Using a process-level allocation method, carbon intensity results were less promising, indicating a net increase in fuel GHG emissions versus petroleum fuels and highlighting the reliance of this scenario on the thermoplastic coproduct.

09 BIOMASS FUELS↗

Microalgae Commodities from Coal-Fired Power Plant Flue Gas CO 2 (Draft Final Report, October 1, 2015 to June 30, 2018)

The Orlando Utilities Commission (OUC) operates an approximately 900-MWe coal fired-power plant at Stanton Energy Center (SEC), which was the site for this project to develop an integrated process for microalgae utilization of flue gas CO 2 for the production of biofuels and animal feeds. Microalgae have both potentially high yields (tons biomass/acre-year) and can be used for biofuels or high value animal feeds. The high biomass yields of microalgae cultures allows efficient use of land and water resources. They can also use wastewaters, recycle fertilizers, and have lower overall greenhouse gas emissions than conventional biofuels or animal feeds. However, many technical issues and technology gaps related to large-scale algae biomass production still must be resolved before such a process can be scaled-up. In this project, techno-economic analyses (TEAs) and life cycle assessments (LCAs) were carried out, projecting the costs and impacts of large-scale algae biomass cultivation for production of biofuels and animal feeds using OUC-SEC flue gas CO 2 . In addition, 3.5-m 2 (~ 40-ft 2 ) experimental raceway ponds were used for the cultivation of microalgae at OUC-SEC and also at the University of Florida. A large-scale algae farm was designed for a 1,300-acre site (~500-ha) located near the OUC-SEC plant, consisting of 400 hectares (1,000 acres) of paddle wheel mixed raceway ponds (each 4 ha), receiving flue gas piped approximately 3 km (2 miles) from the SEC plant. In the first year of this project (Budget Period 1, BP1), the TEA and LCA studies focused on the production of biogas produced from algae biomass and its use to replace coal at the power plant. However, due to the resulting very high cost of CO 2 mitigation, the process was modified to produce compressed natural gas (‘renewable natural gas’, RNG) for transportation fuel from the biogas, in conjunction with wastewater treatment by the algae cultures. During Budget Period 2 (BP2), the TEA and LCA studies focused on the production of animal feeds by the algae farm, using fresh water and agricultural fertilizers. In this case, CO 2 mitigation costs would be entirely covered if the algal biomass could be sold at between a two to three-fold premium above soybean feed, based on a higher nutritional value for the algal biomass. In support of the above analyses, microalgae cultivation experiments were carried out for over two years during this project in small 3.5-m 2 ponds at both SEC and the University of Florida. The most important results were the demonstration at SEC of the cultivation, on flue gas CO 2 , of filamentous algae biomass, which can be harvested at much lower cost than processes required for the colloidal algae species, as currently used by almost all algal mass culture biofuels and animal feeds projects. The yield of biogas production from the algal biomass was investigated by the University of Florida, and the value of the algal biomass as animal feed was projected based on biochemical analysis of the SEC ad UF biomasses. The conclusion from TEA/LCA carried out for the biogas case in BP1 was that a 400-ha algae system producing biomethane for co-firing at OUC-SEC could generate 27,500 MWh/yr of renewable power, offsetting 28,400 Mg CO 2eq /yr, which amounted to less than 1% of the CO 2 emitted by the SEC plant. (Mg = metric ton = 1.10 short tons). The overall cost was projected at $820/Mg of CO 2eq reduction, making this process unfeasible under any possible scenario. More favorable economics were projected when the biogas was converted to higher value transportation fuel, RNG, which also would benefit from renewable fuel (RINs) and possibly other credits. However, only by combining RNG production with municipal wastewater treatment in the algal ponds would CO 2 utilization result in no net cost or even be economically beneficial (i.e., generate income to the power plant). However, in that case the amount of CO 2 utilization would be even lower than that in the biogas case, because a large fraction of the carbon required would be supplied from the wastewater and CO 2 recovered during biogas upgrading to RNG. In brief, use of power plant CO 2 emissions for algal biofuels production was not favorable, even with optimistic assumptions about biomass productivity and biogas yields. For the animal feed case, developed during BP2, algal biomass would replace soybean feeds. The same scale 400-ha algal production plant was projected to produce 26,280 Mg per year of animal feed with a utilization of 45,300 Mg flue gas CO 2 taken up by the algal biomass. However, the process would result in a projected reduction of 112,500 Mg CO 2eq emissions per year, or roughly 1.9% of OUC-SEC emissions, mainly due to the large CO 2eq emissions from conventional soybean production. A major factor in this analysis were greenhouse gas emissions from land use changes. The cost of CO 2 utilization was projected at $116/Mg if the biomass sold for the same price as soybean meal, but if sold as a premium feed (e.g., based on high omega-3 fatty acid and carotenoid content), the power plant could actually charge for the CO 2 utilized. These economic projections are sensitive to assumptions underlying the TEA and LCA models, including biomass composition and the financial models used to project CAPEX and OPEX. Further development of an algal animal feed production using coal-fired power plant flue gas CO 2 is recommended based on the TEA and LCA, as well as experimental results. The required research and development is detailed in a technology gap analysis (Section 11), which focuses on the demonstrations of the cultivation of higher value filamentous algal biomass at OUC-SEC.

01 COAL, LIGNITE, AND PEAT↗

Effect of process variables on the density and durability of the pellets made from high moisture corn stover

A flat die pellet mill was used to understand the effect of high levels of feedstock moisture content in the range of 28–38% (w.b.), with die rotational speeds of 40–60 Hz, and preheating temperatures of 30–110 °C on the pelleting characteristics of 4.8 mm screen size ground corn stover using an 8 mm pellet die. The physical properties of the pelletised biomass studied are: (a) pellet moisture content, (b) unit, bulk and tapped density, and (c) durability. Pelletisation experiments were conducted based on central composite design. Analysis of variance (ANOVA) indicated that feedstock moisture content influenced all of the physical properties at P < 0.001. Pellet moisture content decreased with increase in preheating temperature to about 110 °C and decreasing the feedstock moisture content to about 28% (w.b.). Response surface models developed for quality attributes with respect to process variables has adequately described the process with coefficient of determination (R2) values of >0.88. The other pellet quality attributes such as unit, bulk, tapped density, were maximised at feedstock moisture content of 30–33% (w.b.), die speeds of >50 Hz and preheating temperature of >90 °C. In case of durability a medium moisture content of 33–34% (w.b.) and preheating temperatures of >70 °C and higher die speeds >50 Hz resulted in high durable pellets. It can be concluded from the present study that feedstock moisture content, followed by preheating, and die rotational speed are the interacting process variables influencing pellet moisture content, unit, bulk and tapped density and durability.

09 BIOMASS FUELS↗

A Unified Modeling Framework to Advance Biofuel Production from Microalgae

Modeling efforts to understand the financial implications of microalgal biofuels often assume a static basis for microalgae biomass composition and cost, which has constrained cultivation and downstream conversion process design and limited in-depth understanding of their interdependencies. For this work, a dynamic biological cultivation model was integrated with thermo-chemical/biological unit process models for downstream biorefineries to increase modeling fidelity, to provide mechanistic links among unit operations, and to quantify minimum product selling prices of biofuels via techno-economic analysis. Variability in design, cultivation, and conversion parameters were characterized through Monte Carlo simulation, and sensitivity analyses were conducted to identify key cost and fuel yield drivers. Cultivating biomass to achieve the minimum biomass selling price or to achieve maximum lipid content were shown to lead to suboptimal fuel production costs. Depending on biomass composition, both hydrothermal liquefaction and a biochemical fractionation process (combined algal processing) were shown to have advantageous minimum product selling prices, which supports continued investment in multiple conversion pathways. Ultimately, this work demonstrates a clear need to leverage integrated modeling platforms to advance microalgae biofuel systems as a whole, and specific recommendations are made for the prioritization of research and development pathways to achieve economical biofuel production from microalgae.

09 BIOMASS FUELS↗

Development and validation of time‐domain 1 H‐NMR relaxometry correlation for high‐throughput phenotyping method for lipid contents of lignocellulosic feedstocks

Abstract The bioenergy crops such as energycane, miscanthus, and sorghum are being genetically modified using state of the art synthetic biotechnology techniques to accumulate energy‐rich molecules such as triacylglycerides (TAGs) in their vegetative cells to enhance their utility for biofuel production. During the initial genetic developmental phase, many hundreds of transgenic phenotypes are produced. The efficiency of the production pipeline requires early and minimally destructive determination of oil content in individuals. Current screening methods require time‐intensive sample preparation and extraction with chemical solvents for each plant tissue. A rapid screen will also be needed for developing industrial extraction as these crops become available. In the present study, we have devised a proton relaxation nuclear magnetic resonance ( 1 H‐NMR) method for single‐step, non‐invasive, and chemical‐free characterization of in‐situ lipids in untreated and pretreated lignocellulosic biomass. The systematic evaluation of NMR relaxation time distribution provided insight into the proton environment associated with the lipids in the biomass. It resolved two distinct lipid‐associated subpopulations of proton nuclei that characterize total in‐situ lipids into bound and free oil based on their “molecular tumbling” rate. The T1T2 correlation spectra also facilitated the resolution of the influence of various pretreatment procedures on the chemical composition of molecular and local 1 H population in each sample. Furthermore, we show that hydrothermally pretreated biomass is suitable for direct NMR analysis unlike dilute acid and alkaline pretreated biomass which needs an additional step for neutralization.

pretreatment↗

Analysis of Extracted Ingredients from Dairy Byproduct Using Dimethyl Ether

Whey permeate is a byproduct created in large quantities from the dairy industry with limited applications due to transportation and stability. Whey permeate is primarily comprised of water, carbohydrates, and minerals with minimal contribution of proteins. Various biomasses in food processing have utilized liquid dimethyl ether (DME) as a form of extraction for high-value ingredients and dewatering. DME is a growing interest in wastewater and biomass treatment due to it being a green solvent, ability to dissolve organic solvents, and miscibility with water. The current study is to determine if DME can effectively dewater whey permeate as well as recover components that can be further used in the food industry as a value-added ingredient. Analysis of the liquid extract and solid extract was evaluated with inductively coupled plasma-mass spectrometry (ICP-MS) and solid extracts were further characterized with the application of a scanning electron microscope (SEM).

09 - BIOMASS FUELS↗

Archaeal and bacterial communities across a chronosequence of drained lake basins in arctic alaska

We examined patterns in soil microbial community composition across a successional gradient of drained lake basins in the Arctic Coastal Plain. Analysis of 16S rRNA gene sequences revealed that methanogens closely related to Candidatus ‘Methanoflorens stordalenmirensis’ were the dominant archaea, comprising >50% of the total archaea at most sites, with particularly high levels in the oldest basins and in the top 57 cm of soil (active and transition layers). Bacterial community composition was more diverse, with lineages from OP11, Actinobacteria, Bacteroidetes, and Proteobacteria found in high relative abundance across all sites. Notably, microbial composition appeared to converge in the active layer, but transition and permafrost layer communities across the sites were significantly different to one another. Microbial biomass using fatty acid-based analysis indicated that the youngest basins had increased abundances of gram-positive bacteria and saprotrophic fungi at higher soil organic carbon levels, while the oldest basins displayed an increase in only the gram-positive bacteria. While this study showed differences in microbial populations across the sites relevant to basin age, the dominance of Candidatus ‘M. stordalenmirensis’ across the chronosequence indicates the potential for changes in local carbon cycling, depending on how these methanogens and associated microbial communities respond to warming temperatures.

59 BASIC BIOLOGICAL SCIENCES↗

2023 Billion-Ton Report: An Assessment of U.S. Renewable Carbon Resources - Microalgae

Microalgae is a unique biomass resource that does not need to compete for land and water with other biomass feedstocks because it can be cultivated on low-quality unencumbered land using noncompetitive water types including saline and wastewater. It is included as a complementary resource alongside other biomass feedstocks reported in this study, albeit at higher biomass production costs reflective of more capital-intensive farming operations than typical for terrestrial biomass. Higher biomass costs can be offset by the potential to produce value-added coproducts unique to compositional constituents of microalgae. Relative to the 2016 Billion-Ton Report, this chapter reflects the latest analysis from the 2022 Algae Harmonization Update, which uses the latest parameterized and high-performing saline algal strain, second-generation carbon capture of point-source waste CO 2 , and high-pressure pipeline transport resolved to specific point-source types, saline water sourcing up to 40,000 mg/L total dissolved solids for source and makeup water salinity, blowdown water treatment and recycle, and brine disposal handling. National-scale algal biomass availability potential was calculated at 152 million tons/yr ash-free dry weight (AFDW) (191 million tons/yr dry weight) at an average biomass

09 BIOMASS FUELS↗

Stoichiometric Network Analysis of Cyanobacterial Acclimation to Photosynthesis-Associated Stresses Identifies Heterotrophic Niches

Metabolic acclimation to photosynthesis-associated stresses was examined in the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1 using integrated computational and photobioreactor analyses. A genome-enabled metabolic model, complete with measured biomass composition, was analyzed using ecological resource allocation theory to predict and interpret metabolic acclimation to irradiance, O 2 , and nutrient stresses. Reduced growth efficiency, shifts in photosystem utilization, changes in photorespiration strategies, and differing byproduct secretion patterns were predicted to occur along culturing stress gradients. These predictions were compared with photobioreactor physiological data and previously published transcriptomic data and found to be highly consistent with observations, providing a systems-based rationale for the culture phenotypes. The analysis also indicated that cyanobacterial stress acclimation strategies created niches for heterotrophic organisms and that heterotrophic activity could enhance cyanobacterial stress tolerance by removing inhibitory metabolic byproducts. This study provides mechanistic insight into stress acclimation strategies in photoautotrophs and establishes a framework for predicting, designing, and engineering both axenic and photoautotrophic-heterotrophic systems as a function of controllable parameters.

54 ENVIRONMENTAL SCIENCES↗

Discovering the desirable alleles contributing to the lignocellulosic biomass traits in saccharum germplasm collections for energy cane improvement

Phenotyping Methods: The accessions (which includes 21 taxa and 1,177 accessions) in the World Collection of Sugarcane and Related Grasses (WCSRG) was evaluated for the following traits: arenchyma, internode length and diameter, pubescence, pith, Brix, stalk number and fiber. A core of 300 accessions that included each species in the World Collection was selected by using the Maximization Strategy in MStrat software. Results: The core had a higher diversity rating than random selections of 300 accessions. The Shannon–Weaver Diversity Index scores of the core and whole collection were similar indicating that the majority of the diversity was captured by the core collection. The ranges and medians between the core and WCSRG were similar; only two of the trait medians were not significant at P = 0.05 using the non-parametric Wilcoxon method and the coincidence rate (CR % = 96.2) was high (>80) indicating that extreme values were retained. Thus, the phenotypic diversity of these traits in the WCSRG was well represented by the core collection. Associations Methods: Genotypic and phenotypic data were collected for 1002 accessions of the WCSRG including 209 SSR markers. Association analysis was performed using both General Linear (GLM) and Maximum Likelihood (MLM) models. Different core collections with 300 accessions each were selected based on genotypic, phenotypic and combined data based on the Maximization Strategy in MStrat software. Results: A major portion of the genotyping involving SNPs is being conducted by Dr. Jianping Wang of the University of Florida under the DOE award DE-FG 02-11ER 65214 and the genotypic and phenotypic associations will be reported separately next year. In the current, study forty one and seventeen markers were found to be associated with traits using the GLM and MLM methods respectively including associations with arenchyma, internode length and diameter, pubescence, pith, and Sugar Cane Yellow Leaf Virus. The data indicates that each of the cores and the World Collection are similar to each other genotypically and phenotypically, but the core that was selected using only genotypic data was significantly different phenotypically. This indicates that there is not enough association between the genotypic and phenotypic diversity as to select using only genotypic diversity and get the full phenotypic diversity. Core Collection: Creation and Phenotyping Methods: To evaluate this germplasm for breeding purposes, a representative diversity panel selected from the WCSRG of approximately 300 accessions was planted at Canal Point, FL in three replications. These accessions were measured for stalk height and stalk number multiple times throughout the growing season and Brix and fresh biomass during harvest in 2013 and, stalk height, stalk number, stalk diameter, internode length, Brix and fresh and dry biomass was determined in the ratoon crop harvest in 2014. Results: In correlations of multiple measurements, there were higher correlations for early measurements of stalk number and stalk height with harvest traits like Brix and fresh weight. Hybrids had higher fresh mass and Brix while Saccharum spontaneum had higher stalk number and dry mass. The heritability of hybrid mass traits was lower in the ratoon crop. According to the principal component analysis, the diversity panel was divided into two groups. One group had accessions with high stalk number and high dry biomass like S. spontaneum and the other groups contained accessions with higher Brix and fresh biomass like S. officinarum. Mass traits correlated with each other as expected but hybrids had lower correlations between fresh and dry mass. Stalk number and the mass traits correlated with each other except in S. spontaneum and hybrids in the first ratoon. There were 110 accessions not significantly different in Brix from the commercial sugarcane checks including 10 S. spontaneum accessions. There were 27 dry and 6 fresh mass accessions significantly higher than the commercial sugarcane checks. Core Collection: Fiber analysis Methods: A biomass sample was taken from each accession then shredded and dried. Fiber analysis was then performed on each sample. The acetyl groups, acid insoluble lignin, acid soluble lignin, arabinan, glucan, holocellulose, total lignin, structural ash, and xylan were quantified on a % fiber basis and nonstructural ash on a % total basis. Results: There were significant, but not large differences between species for holocellulose, lignin, acetyl, acid soluble lignin, nonstructural ash, and glucan. For each trait, S. spontaneum had significantly more holocellulose, glucan, lignin, and nonstructural ash and less acetyl and acid soluble lignin than the other species. In all populations, glucan and was positively correlated with holocellulose were positively correlated and glucan and and holocellulose were negatively correlated with lignin. In hybrids, internode length correlated positively with holocellulose and nonstructural ash and negatively with lignin. The heritability estimates for each of the fiber component traits is low indicating that environment is an important factor in fiber composition. Principal component analysis indicated that a large amount of diversity exists within each of the species.

09 BIOMASS FUELS↗

Validation of picogram- and femtogram-input DNA libraries for microscale metagenomics

High-throughput sequencing libraries are typically limited by the requirement for nanograms to micrograms of input DNA. This bottleneck impedes the microscale analysis of ecosystems and the exploration of low biomass samples. Current methods for amplifying environmental DNA to bypass this bottleneck introduce considerable bias into metagenomic profiles. For this study, we describe and validate a simple modification of the Illumina Nextera XT DNA library preparation kit which allows creation of shotgun libraries from sub-nanogram amounts of input DNA. Community composition was reproducible down to 100 fg of input DNA based on analysis of a mock community comprising 54 phylogenetically diverse Bacteria and Archaea. The main technical issues with the low input libraries were a greater potential for contamination, limited DNA complexity which has a direct effect on assembly and binning, and an associated higher percentage of read duplicates. We recommend a lower limit of 1 pg (~100–1,000 microbial cells) to ensure community composition fidelity, and the inclusion of negative controls to identify reagent-specific contaminants. Applying the approach to marine surface water, pronounced differences were observed between bacterial community profiles of microliter volume samples, which we attribute to biological variation. This result is consistent with expected microscale patchiness in marine communities. We thus envision that our benchmarked, slightly modified low input DNA protocol will be beneficial for microscale and low biomass metagenomics.

100 fg↗

Changes in above- versus belowground biomass distribution in permafrost regions in response to climate warming

Permafrost regions contain approximately half of the carbon stored in land ecosystems and have warmed at least twice as much as any other biome. This warming has influenced vegetation activity, leading to changes in plant composition, physiology, and biomass storage in aboveground and belowground components, ultimately impacting ecosystem carbon balance. Yet, little is known about the causes and magnitude of long-term changes in the above- to belowground biomass ratio of plants (η). Here, in this study, we analyzed η values using 3,013 plots and 26,337 species-specific measurements across eight sites on the Tibetan Plateau from 1995 to 2021. Our analysis revealed distinct temporal trends in η for three vegetation types: a 17% increase in alpine wetlands, and a decrease of 26% and 48% in alpine meadows and alpine steppes, respectively. These trends were primarily driven by temperature-induced growth preferences rather than shifts in plant species composition. Our findings indicate that in wetter ecosystems, climate warming promotes aboveground plant growth, while in drier ecosystems, such as alpine meadows and alpine steppes, plants allocate more biomass belowground. Furthermore, we observed a threefold strengthening of the warming effect on η over the past 27 y. Soil moisture was found to modulate the sensitivity of η to soil temperature in alpine meadows and alpine steppes, but not in alpine wetlands. Our results contribute to a better understanding of the processes driving the response of biomass distribution to climate warming, which is crucial for predicting the future carbon trajectory of permafrost ecosystems and climate feedback.

54 ENVIRONMENTAL SCIENCES↗

At-Line Sampling and Characterization of Pyrolytic Vapors from Biomass Feedstock Blends Using SPME-GC/MS-PCA: Influence of Char on Fast Pyrolysis

Solid-phase microextraction (SPME) coupled with GC-MS analysis was used for at-line sampling of pyrolytic vapors produced during fast pyrolysis of biomass. The pure and binary blends of switchgrass (SWG) and pine harvest residues (PT6) were used as feedstock. Sequential SPME sampling allowed for monitoring of changes in the pyrolysis vapors as char accumulated in the fluid bed. The concentration and composition of the vapors desorbed from the SPME fibers were investigated using GC-MS, and the data sets were then analyzed using principal component analysis (PCA) to compare the composition of the pyrolysis vapors over the course of the pyrolysis run. The chemical composition of both carbohydrate and lignin fragments varied as the char builds up in the reactor bed. Fragments derived from cellulose and xylan included anhydrosugars, furans and light oxygenated compounds. Lignin fragments included methoxyphenols, phenolic ketones and aldehydes, low molecular weight aromatics. The composition of the carbohydrate fragments changed more than the lignin fragments as the char build-up in the fluid bed. This combination of SPME-GC/MS-PCA were a novel, easy and effective method for measuring the composition and changes in the composition of pyrolysis vapors during the fast pyrolysis process. Here, this work also highlighted the effect of char build-up on the composition of the overall pyrolysis vapors.

09 BIOMASS FUELS↗

Preserving Aryl Ether Linkages and Higher Yields of Isolated Lignin through Biomass Fibrillation

Ball milling is used for size reduction of biomass for isolating lignin. However, it is known that it can lead to some structural alteration of lignin. Fibrillation using ultrafriction grinding is a newer size reduction technology which was applied to poplar for isolating cellulolytic enzyme lignin. Compositional analysis of poplar, enzymatic hydrolysis yield, water retention value, lignin yield, lignin structural characterization, and lignin molecular weight analysis were carried out to compare lignin isolated from fibrillated and ball milled poplar. By using fibrillation instead of planetary ball milling for size reduction of poplar, higher β-O-4' linkage content, lignin yield, and molecular weight were obtained. Replacing ball milling with fibrillation for size reduction allows recovery of lignin whose structure is closer to that of lignin in biomass.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗