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At least 145 records · Page 8

Corynebacterium glutamicum as an Efficient Omnivorous Microbial Host for the Bioconversion of Lignocellulosic Biomass

Corynebacterium glutamicum has been successfully employed for the industrial production of amino acids and other bioproducts, partially due to its native ability to utilize a wide range of carbon substrates. We demonstrated C. glutamicum as an efficient microbial host for utilizing diverse carbon substrates present in biomass hydrolysates, such as glucose, arabinose, and xylose, in addition to its natural ability to assimilate lignin-derived aromatics. As a case study to demonstrate its bioproduction capabilities, L-lactate was chosen as the primary fermentation end product along with acetate and succinate. C. glutamicum was found to grow well in different aromatics (benzoic acid, cinnamic acid, vanillic acid, and p-coumaric acid) up to a concentration of 40 mM. Besides, 13 C-fingerprinting confirmed that carbon from aromatics enter the primary metabolism via TCA cycle confirming the presence of β-ketoadipate pathway in C. glutamicum . 13 C-fingerprinting in the presence of both glucose and aromatics also revealed coumarate to be the most preferred aromatic by C. glutamicum contributing 74 and 59% of its carbon for the synthesis of glutamate and aspartate respectively. 13 C-fingerprinting also confirmed the activity of ortho-cleavage pathway, anaplerotic pathway, and cataplerotic pathways. Finally, the engineered C. glutamicum strain grew well in biomass hydrolysate containing pentose and hexose sugars and produced L-lactate at a concentration of 47.9 g/L and a yield of 0.639 g/g from sugars with simultaneous utilization of aromatics. Succinate and acetate co-products were produced at concentrations of 8.9 g/L and 3.2 g/L, respectively. Our findings open the door to valorize all the major carbon components of biomass hydrolysate by using C. glutamicum as a microbial host for biomanufacturing.

13C-fingerprinting↗

Using Incremental Changes to Convert Lignocellulosic Feedstocks to Cellulosic Ethanol

One billion tons of biomass feedstocks have been identified for the production of renewable biofuels and biochemicals. This is one of the key carbon feedstocks to supply energy to the transportation sector for light duty, heavy duty and aviation fuels. Utilization of lignocellulosic feedstocks supports an improved energy security by reducing demand of petroleum imports, agricultural development, job creation, and reducing greenhouse gas emissions. To date, however, operational challenges have stymied the industrial production of large volumes of lignocellulosic-based fuels and chemicals. As a result, significant research investment has been led by the United States Department of Energy to understand and improve operational reliability at pioneer cellulosic biorefineries. In this perspective article lignocellulosic conversion technologies are described that have been adopted from the starch ethanol process. The developed process culminated in successful demonstration of 1,000-h integrated runs using several feedstocks, including switchgrass, energy sorghum, and two types of corn kernel fiber. This report highlights process development that solved several of the issues that plagued—and continue to plague—many in the cellulosic sugars space such as biomass feeding into equipment, high ash content, diversified co-product value, and others.

09 BIOMASS FUELS↗

Methodology for assessing the maximum potential impact of separations opportunities in industrial processes

Separation technologies currently used in U.S. manufacturing industries are estimated to account for more than 20% of plant energy consumption. However, accurately determining the impact of new separation technology solutions can sometimes be difficult, especially when evaluating a slate of new candidate separation technologies, each of which has its own separation performance, energy demand, and capital cost. In these cases, a typical approach is to assess each new separation technology by collecting performance and cost information and then using that information to develop a techno-economic analysis to identify overall benefits. While this approach is thorough, it can be time consuming and can hinder reaching a critical understanding of the potential of a given separation challenge, especially when there is no known solution. To address these issues, we developed an assessment methodology, using industrial screening processes, that can be used to better understand the potential impacts of addressing a given separation challenge. This paper presents an overview of our separation challenge stream assessment methodology. The methodology involves defining an “ideal” separator and deriving the associated minimum separation energy. The “ideal” separator represents the most optimistic outlook of a given opportunity so the maximum impact from existing and not-yet-developed solutions can be assessed. Using established biorefinery models, we applied the methodology to 10 different separation challenge streams from two different biomass conversion platforms to identify the type of information that can be obtained. Three of the ten challenge streams assessed had maximum possible cost savings predictions >20%, and associated reductions in process energy carbon intensity ranging from 0 to 54%. Two streams had cost and energy savings potential that were < 5%. Some of the opportunity drivers from the various assessments include higher product yields, reduction or elimination of downstream equipment, new co-products, and cost savings associated with raw materials and energy consumption. The information from these assessments can help guide the selection or development of new separation technology solutions based on the various potential factors that drive the projected benefits.

09 BIOMASS FUELS↗

Engineered poplar for bioproduction of the triterpene squalene

Here we aimed to engineer poplar with optimized pathways to produce squalene, a triterpene commonly used in cosmetic oils, a potential biofuel candidate, and the precursor to the further diversified classes of triterpenoids and sterols. The squalene production pathways were either re-targeted from the cytosol to plastids or co-produced with lipid droplets in the cytosol. Squalene and lipid droplet co-production appeared to be toxic, which we hypothesize to be due to disruption of adventitious root formation, suggesting a need for tissue specific production. Plastidial squalene production enabled up to 0.63 mg/g fresh weight in leaf tissue, which also resulted in reductions in isoprene emission and photosynthesis. These results were also studied through a technoeconomic analysis, providing further insight into developing poplar as a production host

isoprene emission↗

BETO 2021 Peer Review - 1.3.2.001 - Algae Biomass Composition

Addressing critical improvements in biomass productivity and associated biochemical composition is a priority for the economic and sustainable commercial development of biofuels and bioproducts from algae. Capitalizing on pathways that integrate engineering approaches with fundamental biochemistry of photosynthetic organisms will lead to a better understanding of the complex nexus of algae growth rates, productivity and composition. This project focuses on identifying the critical factors for economic development of fuel and bioproduct technologies. Algal compositional characteristics form the foundation of robust economic and business models. This project supports that foundation by developing and validating accurate compositional methods and disseminating them to the greater community. Simultaneously, we build a deep understanding of the dynamic biochemical composition and carbon allocation for biomass value and conversion yields. A co-product portfolio developed under this project demonstrated a 30% increase in intrinsic value. Additionally, an integrated pipeline of molecular diversity mapping for product discovery with quantitative demonstration across species was deployed over the BETO algae program. The advances made here are highly relevant to BETO's multi-year program targets of reducing costs and integrating dynamic biomass composition with conversion processes to provide options for bioproducts, all leveraging the molecular diversity of algae.

algal compositional characteristics↗

Bioconversion of Thermochemical Intermediates

Thermochemical (TC) biofuels production via both pyrolysis and hydrothermal liquefaction produces aqueous waste streams, typically with organic compounds at concentrations of -50-100 g/L. These streams represent a wastewater treatment cost and carbon loss for the TC biorefinery, but the concentration range for these compounds is ideal for bioconversion. To that end, the Bioconversion of Thermochemical Intermediates (BTI) project is developing advanced analytics and engineered microbes to convert these waste streams to co-products, with the overall aim of improving the economics and carbon conversion efficiency of TC biorefining. To date, we have primarily focused on development of advanced analytical chemistry approaches to fully characterize TC aqueous streams and engineering of Pseudomonas putida for conversion of non-conventional substrates, including methylated phenolics, cyclic ketones, furans, and C1-C3 light oxygenates, into atom-efficient products. Two primary challenges are the rapid deployment of aqueous-compatible analytics to changing upstream conditions and dealing with the toxicity of the feed streams to engineered microbes. The project efforts have resulted in engineered strains of P. putida able to consume 90% of the organic compounds in aqueous waste streams from catalytic fast pyrolysis, more than 300-fold toxicity tolerance improvements in P. putida, and carbon closures exceeding 90% for TC wastewater streams across multiple processing technologies.

BASIC BIOLOGICAL SCIENCES,BIOMASS FUELS↗

Cell Free and Immobilization Technologies (CFIT)

Today, several key factors negatively impact the production of fuels and chemicals from renewable sources. Common hindrances in the biological production of biochemicals are: (1) end-product or intermediate toxicity to the microbial biocatalyst, (2) the diversion of carbon to biomass formation, and (3) co-production of undesired byproducts. A particularly attractive alternative is to eliminate the biocatalyst entirely and instead operate the desired metabolic pathways in isolation, thus circumventing the roadblocks of biological toxicity, lower yields, and lack of specificity. However, cell-free enzyme systems still suffer from low productivities owing in part to the effects of free diffusion of intermediates, lack of long term enzyme stability, cofactor cost or inefficient recycling rates, and finally, the cost of enzyme production/purification. This project represents a new effort to propose innovative and cost competitive routes to producing biochemicals from a variety of feedstocks using cell free approaches. These routes will help reduce the current risk and cost associated with classical cell free production. Cell free technologies show promise for application to the production of toxic/inhibitory products or products difficult to separate from microbial growth media and can help reduce the production barriers in multiple areas of biological conversion of feedstocks to biochemicals. More specifically, we are developing new technologies and routes that could be used to produce high value biochemicals such as 2,3 BDO and terpenes (among many others) from biomass derived C5/C6 sugars or lignin but also from waste byproducts such as glycerol. This project will lead to significant innovation and also lead to new concepts and rational design of pathways and enzymes. Within this project, we will develop new metabolic enzyme cascades that will represent natural or artificial combinations of enzymes to produce the desired biochemicals from a variety of feedstocks. We are also developing basic design principals for constructing synthetic metabolons, using fusion proteins and synthetic protein scaffolds, to promote substrate channeling and stability while conserving peak activity. Additionally, our efforts include a techno economic analysis (TEA) of cell free approaches to provide the sensitivities of the process to enzyme loading, activity, pH, reactor volumes, cofactor recycling rates. Finally, we are focusing on further increasing stability, operating lifetime and efficiency of the pathway enzymes by immobilization on support surfaces. We are also focus on immobilizing pathway enzymes or combinations of enzymes on several different conducting polymers and evaluate the effect on stability and operating lifetime. As more combinations become available we will conduct a more systematic study of the means of immobilizing these enzymes. This preliminary work will enable the in-depth study of cofactor recycling at these interfaces using mediators for electron transfer. Taken together, these approaches will enable process intensification, continuous operation, lower capital and separations costs and end-product flexibility, and thus has the potential to contribute significantly to BETO's goals of cost competitive biofuels and bioproducts. To date we have demonstrated 1) the conversion of pyruvate to 2,3 BDO (4 enzymes with cofactor recycling, no additional cofactors needed) at >100g/L (>3g/L/h) using our enzyme tethering approach, 2) Generated mutants of a key redox enzyme with >180- fold improvement in NADH utilization over WT to >85% of NADPH utilization. 3) Produced mevalonate at >10g/L from glucose, limonene at >10g/L from mevalonate and more than >5g/L from glucose with crude enzyme preparations and complete cofactor recycling. 4) successfully generated fully active cross-linked aggregates (CLEAs) of NOX, a cofactor-regenerating water-forming oxidase that is key to our process with increased stability 5) Engineered and identified enzymes able to use the biomimetic cofactor NMN and synthesized new synthetic cofactors.

biocatalysis↗

Evaluating U.S. Natural Gas Environmental Performance

This work summarizes the U.S. Department of Energy National Energy Technology Laboratory's (NETL) modeling and analysis of natural gas environmental performance. Over the past year, NETL updated its U.S. natural gas supply chain life cycle baseline model to incorporate 2020 data, regionalize the transmission and distribution network into six regions, incorporate improved gathering and boosting stage and distribution stage emissions factors, and assign environmental burdens among the various co-products (i.e., crude oil, natural gas liquids, natural gas) of the natural gas supply chain. The updated natural gas model helps generate more comprehensive and regionalized natural gas profiles, mapping production basins to the relevant transmission and distribution regions downstream. The work incorporates data from multiple peer reviewed measurement-based studies to achieve the objective of providing a comprehensive understanding of emissions from the U.S. natural gas supply chain. This presentation also explores future work by NETL to develop a platform that enables stakeholders to customize key parameters within the natural gas LCA model and observe the corresponding effects on the carbon intensity of the overall supply chain. The platform aims to offer a convenient method for stakeholders to obtain relevant insights from the updated model, without diving deep into the intricacies of the NETL natural gas model with hundreds of parameters and linkages.

Khutal, Harshvardhan↗

Pretreatment of Algae for the Production of Fuels and Chemical Coproducts

Algal biomass is a promising resource for producing renewable fuels and chemicals, but despite decades of research, algal biorefining remains in a pre-commercial state. Recent economic analyses have indicated that high-value co-products are necessary to offset the cost of fuel production, implying the need for algae fractionation technology. One promising fractionation approach is Combined Algal Processing (CAP), which pretreats and extracts algal biomass to produce a fermentable aqueous hydrolysate, an organic lipid fraction, and a residual solids fraction. The CAP approach has historically employed a dilute acid pretreatment to lyse cells and solubilize algal carbohydrates for fermentation of the hydrolysate phase, but we recently identified alternative pretreatments with the potential to decrease costs and environmental impacts, including different implementations of dilute acid pretreatment, dilute alkali pretreatment, enzymatic pretreatment, and flash hydrolysis. We conducted a screening of six different pretreatment approaches across nine algae strains of varying composition, and measured pretreatment effectiveness as a combination of carbon and nitrogen solubilization, lipid extraction yield, and lipid speciation. In this screening, we found traditional dilute acid pretreatment to provide the most robust pretreatment performance, though other pretreatments were competitive, and some performed better for certain strains of algae. These results highlight the interplay between algae composition and pretreatment effectiveness.

algae↗

Biomass Attributes and Attribute Modifications Affecting Systems and Methods to Separate and Fractionate. In: Handbook of Biorefinery Research and Technology

Chemical and physical heterogeneity in biomass feedstocks such as agricultural or forestry residues is due to substantial differences in plant tissue types. These differences can contribute significant challenges to handling, preprocessing, and conversion in biorefining processes. An understanding of this chemical and physical heterogeneity can be used to inform fractionation technologies that could facilitate more streamlined processing and potentially be employed to yield multiple co-product streams for a single feedstock. In this chapter, the motivation and scope of biomass fractionation is first outlined. Physical and chemical properties of biomass feedstocks, along with their distribution and diversity within plants is next discussed with respect to how these differences can be exploited in a fractionation process. A summary of some of the key physical principles that allow for fractionation is next covered along with how these physical principles are exploited in equipment designs. Examples from the literature are briefly discussed that highlight how these approaches can be employed to achieve processing objectives. Several case studies on physical fractionation of corn stover and forestry residues are presented that illustrate how integrated fractionation processes could be employed. Finally, prospects and potential economic drivers for adoption of biomass fractionation technologies are discussed.

Biomass chemical properties↗

Evolution of a Geological Model for Co-Producing Electricity at the Blackburn Oil Field, Nevada: Preprint

Oil fields characterized by significant proportions of hot water in the reservoir offer a unique opportunity for geothermal energy co-production and utilization. Development of an accurate geological model is essential in understanding the subsurface. However, this is a challenge at the preliminary stages of resource assessment when data on the subsurface parameters and subterranean fault network is scarce. Our focus in this paper is to explore a methodology for developing an accurate conceptual geological model of the complexly faulted reservoir at the Blackburn Field, Nevada, to create a numerical simulation that will eventually be used to assess the potential of co-producing electricity. There are four steps to the evolution of this model: (1) collection of public data and research on the geological setting of the system; (2) digitization of seismic time-interpretation and cross-sections to create the most representative model of the subsurface; and (3) creation of a natural-state numerical model to simulate the thermal gradient through the subsurface. Utilizing publicly available data of the geological system and Leapfrog conceptual modeling software, we refined a preliminary model to be more representative of the subsurface geology of an active geothermal exploration stage project. The conceptual geological model developed has been imported into VOLSUNG reservoir modeling software to create a natural-state model, further characterizing the Blackburn Field reservoir's thermal gradient. In the future, new seismic data will be utilized to further evolve the conceptual geological model for numerical simulations.

Basin and Range province↗

Herbaceous Feedstock 2022 State of Technology Report

The U.S. Department of Energy promotes production of advanced liquid transportation fuels from lignocellulosic biomass by funding fundamental and applied research that advances the state of technology (SOT). As part of its involvement in this mission, Idaho National Laboratory completes an annual SOT report for nth-plant and 1st-plant herbaceous biomass feedstock logistics. The purpose of the SOT is to provide the status of feedstock supply system technology development for herbaceous biomass to biofuels relative to technical targets and cost goals from specific design cases, based on data and experimental results. Although conventional feedstock supply systems form the backbone of the emerging biofuels industry, they have limitations that restrict widespread implementation on a national scale. To meet the demands of the future industry, the feedstock supply system must shift from the conventional system to what has been termed “advanced” supply systems. In advanced designs, a distributed network of aggregation and processing centers, termed “depots,” are employed near the points of biomass production (i.e., the field or forest) to reduce feedstock variability and produce feedstocks of a uniform format, moving toward biomass commoditization. The 2022 Herbaceous SOT is part of a vision of achieving an implemented advanced feedstock supply system, which produces a stable, tradable commodity at the decentralized distributed depot. It utilizes feedstock fractionation by incorporating technologies that can separate the biomass into its anatomical fractions (leaves, husks, stems and cobs) to reduce impurities and produce fractions that satisfy downstream quality considerations. By using a series of air classification steps, this strategy can reduce the extrinsic ash in corn stover and produce enriched tissue fractions that can be blended to a conversion specification or converted individually in optimized biochemical conversion campaigns. Additionally, a majority of the leaves (which do not meet the quality specification) are separated out early and can be supplied to alternate markets. The 2022 Herbaceous SOT incorporates an advanced biomass fractionation and processing system to produce pellets enriched tissues from three-pass corn stover. The resulting enriched pellets are delivered to the biorefinery individually where they can be blended to a specification or converted in campaigns where the conditions are optimized for each tissue. Unused fractions can be sent to a a midstream market or to a different conversion process that is better suited to their properties to offset the cost of the delivered feedstock. The main benefits from the proposed system can be summarized as: (1) $6.86/dry ton (2016$) lower cost for the air classification due to elimination of the requirement to discard the high ash lights fraction; (2) $1.56/dry ton lower delivered cost by selling the unsuitable leaf fraction into the feed market as a midstream co-product (assuming a selling price that is 11% higher than their cost of production); (3) 0.98% increase in carbohydrate content (from 60.16% to 61.14%); and (4) 0.97% decrease in ash content (from 6.00% to 5.03%) compared to the 2021 Herbaceous SOT. Overall, the 2022 nth-plant Herbaceous SOT predicts a modeled delivered feedstock cost of $78.64/dry ton (2016$) if it is assumed that the enriched leaf fraction is sold at its production cost; this is a slight increase of $0.43/dry ton increase from the 2021 Herbaceous SOT nth-Supply case cost. The increased cost derived from a $0.38/dry ton increase in transportation and handling cost to procure more biomass (to replace the enriched leaf fraction that was not delivered to the biorefinery. The total preprocessing cost was $0.27/dry ton higher than the 2021 result because of updates to energy consumption, purchasing price and dry matter loss data for the rotary shear ($3.00/dry ton increase) and the pelleting mill ($4.52/dry ton increase). The data utilized were generated in pilot-scale tests in the Biomass Feedstock National User Facility (BFNUF) at INL and at Forest Concepts, including tests for rotary shear and pelleting of the air classified fractions. A greenhouse gas emissions analysis was performed by Argonne National Laboratory using the most up to date version of the Greenhouse Gases, Regulated Emissions, and Energy use in Transportation model (GREET®). The analysis showed an increase of 17.34 kg CO2e/dry ton from the 2021 SOT (67.71 kg CO2e/ton in the 2021 Herbaceous SOT to 85.05 kg CO2e/ton in the 2022 Herbaceous SOT). The net increase is primarily attributed to increased energy consumption in pelleting mill.

09 BIOMASS FUELS↗

Advancing Geothermal Research: Fiscal Year 2025 Accomplishments Report

This is a summary of geothermal work done at the National Renewable Energy Laboratory (NREL) in Fiscal Year 2025. This year brought increased attention to the geothermal industry and NREL's geothermal research portfolio. With more than 70 active projects, NREL research spanned the areas of resource exploration and characterization; conventional and next-generation geothermal technologies; subsurface thermal energy storage; heating and cooling; co-production of geothermal with critical minerals and oil and gas; modeling and analysis leveraging expertise in data science and machine learning; and more.

15 GEOTHERMAL ENERGY↗

Advancing Geothermal Research: Fiscal Year 2025 Accomplishments Report

This is a summary of geothermal work done at the National Laboratory of the Rockies in Fiscal Year 2025. This year brought increased attention to the geothermal industry and NLR's geothermal research portfolio. With more than 70 active projects, NLR research spanned the areas of resource exploration and characterization; conventional and next-generation geothermal technologies; subsurface thermal energy storage; heating and cooling; co-production of geothermal with critical minerals and oil and gas; modeling and analysis leveraging expertise in data science and machine learning; and more.

15 GEOTHERMAL ENERGY↗

Mind the gaps! Climate scientists should heed lessons in collaborative storytelling from William Shakespeare

Abstract To co‐produce locally relevant climate knowledge, climate scientists are engaging in new collaborations—with stakeholders and with scholars in the humanities, arts, and social sciences. In our work as a Shakespeare scholar‐turned‐public‐humanist and a climate scientist, we have created a methodology that allows researchers and communities to co‐produce knowledge by co‐producing narratives. We combine principles from emerging climate “storylines” research with collaborative storytelling inspired by William Shakespeare's plays and theatrical practices. Shakespeare's plays spark collaborations and interpretations, in part, because of how Shakespeare leaves gaps in the narrative. These gaps allow others to enter as collaborators, creating a “cognitive ecology” that fosters knowledge and action among all engaged. Integrating these methods into climate storyline‐making offers a radical paradigm: it upends the scientist's role as the focal storyteller and expert, and fosters, instead, partnership, equity, and a co‐exploration of multiple uncertainties. It is time for researchers to cede control to a cognitive ecology of collaborative action. This article is categorized under: Trans‐Disciplinary Perspectives > Humanities and the Creative Arts

54 ENVIRONMENTAL SCIENCES↗

Optimized production of a bioactive human recombinant protein from the microalgae Chlamydomonas reinhardtii grown at high density in a fed-batch bioreactor

Microalgae have been identified as an alternative platform to produce high-quality biomass and subsequent bioproducts, such as foods, feeds, nutritional supplements, recombinant proteins, and biofuels. Traditional biotechnological hosts for therapeutic proteins, such as the bacteria Escherichia coli and mammalian CHO cells, have long been established as the dominate platforms, but recent advances have shown that microalgae can potentially serve as an alternative platform. In the present study, we examine the potential of the microalga Chlamydomonas reinhardtii to produce a complex human recombinant protein in a high-density heterotrophic culture. The recombinant human protein, ICAM-1, was targeted for secretion to the extracellular media of the culture from cells grown in a bioreactor using a fed-batch strategy to achieve high cell density. Ultimately, this resulted in a maximum biomass titer of 40 g/L and a recombinant protein titer of 50 mg/L. The algal-produced ICAM-1 protein showed comparable bioactivity to mammalian cell culture produced ICAM-1, as measured using binding assays for its native ligand LFA-1. This work shows that C. reinhardtii is a viable option to produce complex recombinant proteins, with native biological activity, at high concentrations using a fed batch heterotrophic growth strategy.

60 APPLIED LIFE SCIENCES↗

A research agenda for the science of actionable knowledge: Drawing from a review of the most misguided to the most enlightened claims in the science-policy interface literature

Linking science with action affords a prime opportunity to leverage greater societal impact from research and increase the use of evidence in decision-making. Success in these areas depends critically upon processes of producing and mobilizing knowledge, as well as supporting and making decisions. For decades, scholars have idealized and described these social processes in different ways, resulting in numerous assumptions that now variously guide engagements at the interface of science and society. We systematically catalog these assumptions based on prior research on the science-policy interface, and further distill them into a set of 26 claims. We then elicit expert perspectives (n = 16) about these claims to assess the extent to which they are accurate or merit further examination. Out of this process, we construct a research agenda to motivate future scientific research on actionable knowledge, prioritizing areas that experts identified as critical gaps in understanding of the science-society interface. The resulting agenda focuses on how to define success, support intermediaries, build trust, and evaluate the importance of consensus and its alternatives – all in the diverse contexts of science-society-decision-making interactions. We further raise questions about the centrality of knowledge in these interactions, discussing how a governance lens might be generative of efforts to support more equitable processes and outcomes. We offer these suggestions with hopes of furthering the science of actionable knowledge as a transdisciplinary area of inquiry.

99 GENERAL AND MISCELLANEOUS↗