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At least 811 records · Page 45

Yeast cells having disrupted pathway from dihydroxyacetone phosphate to glycerol

Yeast cells are genetically modified to disrupt a native metabolic pathway from dihydroxyacetone to glycerol. In certain aspects, the yeast cell is of the genera Kluyveromyces, Candida or Issatchenkia. In other aspects, the yeast cell is capable of producing at least one organic acid, such as lactate. The yeast cells produce significantly less glycerol than the wild-type strains, and usually produce greater yields of desired fermentation products. Yeast cells of the invention often grow well when cultivated, despite their curtailed glycerol production.

Dundon, Catherine Asleson↗

Microorganism with knock-in at acetolactate decarboxylase gene locus

Provided herein is a genetically engineered microorganism comprising knock-in of DNA at an acetolactate decarboxylase gene locus. Replacement of the acetolactate decarboxylase gene with DNA encoding one or more native or nonnative enzymes confers certain advantages, including fermentation stability and increased production of native and nonnative products from gaseous substrates.

Leang, Ching↗

Coupling Waste Feedstocks to Microbial Protein for a Circular Food System

The global food system is responsible for approximately 34% of annual greenhouse gas (GHG) emissions and up to 85% of water consumption. This critical sector suffers from intensive and inefficient land and water use, the generation of multiple (solid, liquid, and gaseous) waste streams, and high fuel, fertilizer, and pesticide consumption. The production of waste-derived microbial protein (MP) represents a promising alternative for reducing the environmental impacts of protein production relative to conventional agriculture. MP can be mass-produced in volumetrically scalable cultivation processes on short timescales, enabling facile up-scaling with lower greenhouse gas emissions, land use, and water impacts than animal and, in some cases, plant protein production. MP can also be produced from waste feedstocks, diverting waste from landfills or the natural environment. Here, we present the availability and suitability of waste feedstocks for MP production, as well as the fermentation and downstream processes required to convert MP into human food products. We discuss the challenges and opportunities facing waste-derived bacterial MP and highlight key areas for innovation in both the microbiology and process design space for a more sustainable and circular food system.

BASIC BIOLOGICAL SCIENCES,ENERGY PLANNING, POLICY,↗

Adaptive Laboratory Evolution for Enhanced Performance of Cupriavidus necator on Formic Acid

The need to decarbonize our economy is becoming ever more pressing. CO2, can be electrochemically reduced to formic acid, a soluble C1 molecule that can be used to store carbon and energy. Cupriavidus necator H16, a soil bacterium, capable of consuming and growing on formic acid as its sole carbon source, is well positioned to upgrade CO2-derived formic acid into value added chemicals such as sustainable aviation fuel. To improve the performance of C. necator on formic acid adaptive laboratory evolution (ALE), a proven tool for improving microbial fitness, has been conducted using continuous pH-stat bioreactors. The system works on the basis that consumption of formic acid raises the pH and triggers the addition of more formic acid to maintain the pH at 6.7, such that formic acid is provided at the same rate as it is consumed. This system has been coupled with level control to achieve continuous fermentation where cells acquiring mutations that improve growth on formic acid become more abundant in the population, from which they can be isolate and characterized. This configuration where the dilution rate is controlled by pH-stat has been found to seek out a dilution rate that matches the exponential growth rate of the organism. During developmental experiments it was discovered that formic acid accumulated to inhibitory levels. It was determined that the nitrogen source, ammonium hydroxide, must be tailored to the carbon consumption to avoid formic acid accumulation. ALE has run in three lineages for 2000+ hours and 300+ generations. Growth of evolved isolates derived from all lineages will be evaluated on formic acid and those with improved growth rates or biomass yield will be subjected to whole genome sequencing to identify potentially causative mutations. These mutations will be evaluated individually and in combination to identify those that improve growth on formic acid.

adaptive laboratory evolution↗

Zeolitic catalytic conversion of alcohols to hydrocarbons

A method for converting an alcohol to a hydrocarbon, the method comprising contacting said alcohol with a metal-loaded zeolite catalyst at a temperature of at least 100° C. and up to 550° C., wherein said alcohol can be produced by a fermentation process, said metal is a positively-charged metal ion, and said metal-loaded zeolite catalyst is catalytically active for converting said alcohol to said hydrocarbon.

Narula, Chaitanya K.↗

Adaptive Laboratory Evolution for Enhanced Performance of Cupriavidus Necator on Formic Acid

The threat of global warming, driven by rising carbon emissions, highlights the need to decarbonize our economy. This requires innovative solutions for managing carbon waste and its effective utilization. One promising method for CO2 capture and sequestration is the electrochemical reduction of CO2 to formic acid, a soluble C1 molecule that can be used to store carbon and energy, and as a feedstock for biological conversion. Cupriavidus necator H16, a soil bacterium capable of consuming and growing on formic acid as its sole carbon and energy source, is well positioned to upgrade CO2-derived formic acid into platform chemicals and fuel precursors. To improve the performance of C. necator on formic acid, adaptive laboratory evolution (ALE), a proven tool for improving microbial fitness, has been conducted using continuous pH-stat bioreactors. The system works on the basis that consumption of formic acid raises the pH and triggers the addition of more formic acid to maintain the pH (in this case 6.7), such that formic acid is provided at the same rate as it is consumed. This system has been coupled with level control to achieve continuous fermentation where cells acquiring mutations that improve growth on formic acid become more abundant in the population, from which they can be isolated and characterized. During developmental experiments it was discovered that formic acid accumulated to inhibitory levels. It was determined that the nitrogen source, ammonium hydroxide, must be tailored to the carbon consumption to avoid formic acid accumulation. The ALE ran in three lineages for approximately 3000 hours and more than 500 generations. Evolved isolates obtained from each lineage demonstrated an increase in growth rate in conjunction with improve formate utilization compared to the parental strain when evaluated in pH-stat bioreactors. The isolates with improved performance were then subjected to whole genome sequencing to identify potentially causative mutations. Mutations in several key genes across different lineages have been found and will be evaluated individually and in combination to identify those that improve growth on formic acid. Incorporating these mutations into production strains has the potential to greatly improve formic acid conversion and further industrial decarbonization.

adaptive laboratory evolution↗

Evaluating Impacts of Sustainable Aviation Fuel Production with CO2-to-Fuels Technologies on High Renewable Share Power Grids

This paper investigates the impact of Sustainable Aviation Fuel production using CO2-to-Fuels technologies on a future power grid with a high share of renewable energy. We focus on understanding the implications of the 2050 SAF production goal on the U.S. power system's long-term planning, encompassing generation, transmission, and cost analysis. Via the Regional Energy Deployment System (ReEDS) model, we developed a detailed SAF electricity demand model based on a low-temperature electrolysis-syngas fermentation-ethanol pathway. Four SAF target scenarios which aims to meet 10%, 15%, 20%, and 27% of SAF demand by 2050. These scenarios are exhaustively simulated to assess their impact on the power grid. Our results reveal that increasing SAF demand will result in higher electricity requirements, as well as expanded generator and transmission capacities, leading to an overall rise in system costs. However, these impacts are manageable within the broader context of U.S. capacity expansion plans. This study provides valuable insights into incorporating the CO2-to-Fuels electricity demand model and other carbon capture technologies in power system planning, emphasizing their significance in shaping a sustainable energy future.

BIOMASS FUELS,POWER TRANSMISSION AND DISTRIBUTION↗

Evaluating Impacts of Sustainable Aviation Fuel Production with CO2-to-Fuels Technologies on High Renewable Share Power Grid: Preprint

This paper investigates the impact of Sustainable Aviation Fuel (SAF) production using CO2-to-Fuels technologies on a future power grid with a high share of renewable energy. We focus on understanding the implications of the 2050 SAF production goal on the U.S. power system's long-term planning, encompassing generation, transmission, and cost analysis. Via the Regional Energy Deployment System (ReEDS) model, we developed a detailed SAF electricity demand model based on a low-temperature electrolysis-syngas fermentation-ethanol pathway. Four SAF target scenarios which aimto meet 10%, 15%, 20%, and 27% of SAF demand by 2050. These scenarios are exhaustively simulated to assess their impact on the power grid. Our results reveal that increasing SAF demand will result in higher electricity requirements, as well as expanded generator and transmission capacities, leading to an overall rise in system costs. However, these impacts are manageable within the broader context of U.S. capacity expansion plans. This study provides valuable insights into incorporating the CO2-to-Fuels electricity demand model and other carbon capture technologies into power system planning, emphasizing their significance in shaping a sustainable energy future.

BIOMASS FUELS,ENERGY PLANNING, POLICY, AND ECONOMY↗

Decarbonizing Hydrogen Production: Assessing A Net-Negative Pathway

Hydrogen is gaining prominence as a key factor in the world's transition to a cleaner energy future. The International Energy Agency (IEA)'s Global Hydrogen Review 2023 reports that the number of low-emission hydrogen production projects is increasing rapidly. The potential for growth in new applications such as heavy industry, transportation, and power generation is significant. The IEA urges more decisive action to spur demand for low-emission hydrogen to achieve climate goals. While hydrogen is produced through various industrial methods, each with its own advantages and disadvantages, low-carbon hydrogen is critical for mitigating climate change and is incentivized by the Clean Hydrogen Production Tax Credit (45V). To this end, we have evaluated a commercial technology that can produce low- or negative-carbon hydrogen via ethanol catalytic oxidative reforming. This study assessed life cycle greenhouse gas emissions (carbon intensity or CI) associated with the hydrogen production technology. A total of 24 scenarios were evaluated, encompassing (a) Gen1 versus Gen2 ethanol inputs, (b) carbon capture and sequestration (CCS) of upstream fermentation CO2, and (c) oxygen sourcing via air separation unit (ASU) versus purchased or on-site production of oxygen as a byproduct of hydrogen electrolysis with a proton exchange membrane (PEM). Key findings include that the base case CI for hydrogen production using Gen2 ethanol from corn stover is lower than Gen1 dry mill corn ethanol. The study also points out that the CI for hydrogen production using PEM-O2 is lower than that using ASU-O2, whether the PEM-O2 is produced on-site or off-site (importing). When sourcing oxygen from on-site PEM-O2, the Gen1 and Gen2 ethanol-derived hydrogen exhibit favorable net-negative CI values for all evaluated scenarios, especially if the upstream ethanol CCS is included. As a reference, the 45V regulatory threshold for generating clean hydrogen tax credits is a CI below 0.45 kg CO2e/kg hydrogen.

BIOMASS FUELS,HYDROGEN↗

Microorganism with knock-in at acetolactate decarboxylase gene locus

Provided herein is a genetically engineered microorganism comprising knock-in of DNA at an acetolactate decarboxylase gene locus. Replacement of the acetolactate decarboxylase gene with DNA encoding one or more native or nonnative enzymes confers certain advantages, including fermentation stability and increased production of native and nonnative products from gaseous substrates.

Leang, Ching↗

Chapter 3: New Developments on Ionic Liquid-Tolerant Microorganisms Leading Toward a More Sustainable Biorefinery

The growing concerns about climate change and energy security are driving the development of bio-based technologies to produce renewable liquid fuels and chemicals. Ionic liquids (ILs) have demonstrated to be promising solvents to pretreat lignocellulosic residues, promoting efficient enzymatic hydrolysis of lignocellulosic carbohydrates into sugars, which can be further used by microorganisms to produce biofuels and other value-added chemicals. Despite their unique properties to effectively deconstruct plant cell walls, ILs show strong interactions with the pretreated biomass, and their presence is often inhibitory to cellulolytic enzymes and microorganisms. The most advanced biorefinery concepts based on IL pretreatments focus on the development of more biocompatible ILs and more robust microbial strains with higher tolerance to ILs. This chapter provides an overview and a discussion over the main efforts performed on the screening and development of IL-tolerant microbial strains, as well as in more biocompatible IL pretreatment methods. These early research advancements in this field offer a baseline and a platform for future research with the goal of improving the sustainability and economic viability of IL pretreatment-based biorefineries.

bioconversion↗

Functional plasticity of HCO 3 – uptake and CO 2 fixation in Cupriavidus necator H16

Despite its prominence, the ability to engineer Cupriavidus necator H16 for inorganic carbon uptake and fixation is underexplored. We tested the roles of endogenous and heterologous genes on C. necator inorganic carbon metabolism. Deletion of β-carbonic anhydrase can had the most deleterious effect on C. necator autotrophic growth. Replacement of this native uptake system with several classes of dissolved inorganic carbon (DIC) transporters from Cyanobacteria and chemolithoautotrophic bacteria recovered autotrophic growth and supported higher cell densities compared to wild-type (WT) C. necator in batch culture. Strains expressing Halothiobacillus neopolitanus DAB2 (hnDAB2) and diverse rubisco homologs grew in CO 2 similarly to the wild-type strain. Our experiments suggest that the primary role of carbonic anhydrase during autotrophic growth is to support anaplerotic metabolism, and an array of DIC transporters can complement this function. This work demonstrates flexibility in HCO 3 - uptake and CO 2 fixation in C. necator, providing new pathways for CO 2 -based biomanufacturing.

59 BASIC BIOLOGICAL SCIENCES↗

Overcoming barriers to medium-chain fatty alcohol production

Medium-chain fatty alcohols ( mc FaOHs) are aliphatic primary alcohols containing six to twelve carbons that are widely used in materials, pharmaceuticals, and cosmetics. Microbial biosynthesis has been touted as a route to less-abundant chain-length molecules and as a sustainable alternative to current petrochemical processes. Several metabolic engineering strategies for producing mc FaOHs have been demonstrated in the literature, yet processes continue to suffer from poor selectivity and mc FaOH toxicity, leading to reduced titers, rates, and yields of the desired compounds. Here, this opinion examines the current state of microbial mc FaOH biosynthesis, summarizing engineering efforts to tailor selectivity and improve product tolerance by implementing engineering strategies that circumvent or overcome mc FaOH toxicity.

59 BASIC BIOLOGICAL SCIENCES↗

Interactive effects of precipitation, temperature, and grazing pressure on the net ecosystem carbon balance of grazing lands across the continental United States

Grazing lands cover approximately one-third of the contiguous United States, support much of the nation's beef production, and are an important component of the U.S. terrestrial carbon budget. In this study, we quantified net ecosystem carbon balance (NECB), the net status of grazing lands as a carbon sink (C-sink) or source (C-source) and a key determinant of soil health and productivity. Our primary objective was to synthesize multiple years of annual NECB across heterogeneous grazing lands across the continental U.S and evaluate annual NECB against physical drivers (mean annual precipitation (MAP), mean annual temperature (MAT), vegetation, and moisture condition) and management practices (grazing pressure index (GPI) and fertilization history). We hypothesized that (1) NECB is higher in mesic and fertilized grasslands; (2) NECB increases with MAP and MAT but decreases with GPI; and (3) interactive effects exist among MAP, MAT, and GPI. Using carbon fluxes measured by eddy covariance towers and methane emissions including both enteric methane and manure derived from stocking rates across seven USDA Long-term Agroecosystem Research Network sites, we found: (1) grazing lands were a C-sink or carbon neutral at most sites; (2) vegetation type, moisture conditions, or fertilization had no significant effect on NECB; (3) NECB increased with MAP and MAT, but decreased with a higher GPI; and (4) MAT had a significant positive effect on NECB when MAP exceeded 750 mm (greater water availability). The effect of GPI on NECB was significantly negative when MAP was below 1000 mm, significantly negative when MAT < 12 °C and significantly positive when MAT > 16 °C. Thus, most grazing lands in our study acted as C-sinks unless water deficit, low temperature, or heavy grazing were interactively present. Understanding how climate and management influence NECB of grazing lands is key to maintaining resilient agroecosystems that secure beef production and sustain rural prosperity.

LTAR↗

Enhancing methane production of anaerobic digested sewage sludge by advanced wet oxidation & steam explosion pretreatment

Anaerobic digestion (AD) of sewage sludge is associated with lowered carbon conversion efficiency due to the recalcitrance of microbial cells in the Waste Activated Sludge (WAS) from the aeration basins with 50 % of the dry weight still being present after AD. The effect of Advanced Wet Oxidation & Steam Explosion (AWOEx) pretreatment on the methane yield at variable temperatures (165–200 °C), residence time (15–45 min) and oxygen dosage (1%–10% based on VS concentration) was tested in this study. Central composite design (JMP® Pro) was used to design 19 runs of AWOEx pretreatment of dewatered digested sewage sludge (DADSS) at different operating conditions. After pretreatment the DADSS was fed to 19 bioreactors operated in semi-continues mode at 37 °C. The highest average methane yields of 183 mL/g VS and 170 mL/g VS were found for the bioreactors receiving DADSS pretreated at 165 °C with a retention time of 15 min and 10 % O2 (Condition 1), and, 182.5 °C with a retention time of 15 min and 5.5 % O2 (Condition 2). This corresponds to an increase in the methane production of 156.2 % and 140.5 % compared to the methane production from the DADSS without pretreatment.

09 BIOMASS FUELS↗